0191 2817a PDF

QUANTITATIVE
PHARMAOEUTICAL,
OHEMISTRY
CONTAINING THEORY AND PRACTICE OF
QUANTITATIVE ANALYSIS APPLIED
TO PHARMACY
BY
GLENN L. JENKINS, PH.D.

Profes80r of Pharmaceutical Chemi8try, Colle(Jo of Pharmacy,
Uni.eraity of Minnesota
AND
ANDREW G. DuMEZ, PH.D.

Profe8sor of Pharmacy and Dean of the School of Pharmacy,
Univeraity of Maryland
SECOND EDITION
McGRAW-HILL BOOK COMPANY,

NEW YORK AND LONDON
1937
INC.
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McGRAW-HILL PUBLICATIONS IN PHARMACY
QUANTITATIVE PHARMACEUTICAL
CHEMISTRY
McGRAW-HILL PUBLICATIONS IN
PHARMACY
A
SERIES OF TEXTS AND REFFERENCE WORKS

OUTLINED BY THE FOLLOWING COMMITTEE
C. W. JOHNSON, Chairman,
Dean, University of Washington, College of Pharmacy.
EDWARD SPEASE, Acting Chairman and Consulting Ed7.'tOT,
Dean, Western Reserve University, School of Pharmaoy.
JOHN
G.
BEARD,
Professor of Pharmacy, University of North Carolina,

School of Pharmacy.
W. W. OHARTERS,
Direqtor of the Bureau of Educational Research, Ohio
State University. Recently Director in Charge of the
Commonwealth Study of Pharmacy.
H. C. CHRISTENSEN,
Secretary National Assooiated Boards of Pharmacy,
Chicago, Illinois.
ZADA M. COOPER,
Associate Professor of Pharmaoy, State University of

Iowa, College of Pharmacy.
ANDREW G. DuMEZ,
Dean, University of Maryland, School of Pharmaoy.
CLAIR
A.
DYE,
D'l"n, Ohio State University, College of Pharmaoy.

D. B.
R.
JOHNSON,
Dean, University of Oklahoma, Sohool of Pharmacy.

C. B. JORDAN,
Dean, Purdue Uiiiver~ity, School of Pharmacy.
TOWNES
R.
LEIGH,
Dean, University of Florida, School of Pharmacy.

RUFus A. LYMAN.
Dean. University of Nebraska, College of Pharmaoy.
WORTLEY
F.
RUDD,
Dean, Medical College of Virginia. Sohool of Pharmaoy.

CHAS.
H.
STOCKING,
Associate Professor of Pharmaoy, University of Michigan, College of Pharmaoy.

ARNO VIEHOEVER t
Profeasor of Biology and Pharmacognosy, Philadelphia

College of Pharmacy.
A.
I. WINNE,
Secretary, Virginia State Board of Pharmacy, Richmond.
Virginia.
L.
McGRAW -HILL
PUBLICATIONS IN PHARMACY
Jenkin. and DuM ez-QUANTITATIVE PHARMACEUTICAL CHEMISTRY
JordanQUALITATIVE ANALYSIS FOR STUDENTS OF PHARMACY AND MEDICINE
LynnPHARMACEUTICAL THERAPEUTICS
Speaa_
PHARMACEUTICAL MATHEMATICS
COPYRIGHT,
1931, 1937,
BY THE
MCGRAW-HILL BOOK COMPANY, INC.

PRINTED IN THE UNITED STATES OF AMERICA
All rights reserved. This book, or

parts thereof, may not be reproduced
in any form without permission of
the publishers.
THE
USE
IN
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VOLUM!::
OF
CERTAIN
PORTIONS
OF THE
TEXT OF THE UNITED STATES PHARMACOPOEIA IS ElY VIRlUE

OF
PERMISSION
RECEIVEO
FROM
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OF THE UNITED STATES PHARMACOPOEIAL. CONVENTION.
SAID BOARD
OF TRUSTEES
INACCURACY OF
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STATEMENT
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FOR ANY ERRORS IN THE
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STRENGTHS.
PERMISSION TO USE FOR COMMENT PARTS OF' THE TEXT OF'
THE NATIONAL FORMULARY, SIXTH EDITION, IN THIS VOLUME

HAS BEEN GRANTED BY THE COMM1TTEE: ON PUBLICATIONS By
THE AUTHORITY OF THE COUNCIL OF 'l'HE AMERICAN PHARM4._

CEUTICAL
ASSOCIATION~
THE MAPLE PRESS COMPANY, YORK, PA.
PREFACE TO THE SECOND EDITION

Since the publication of the first edition of this book, numerous
changes have been made in the official methods of analysis of
pharmaceutical materials in the United States Pharmacopoeia
XI and in the National Formulary VI. The detailed procedures
and discussions have been changed or rewritten in the present
edition to conform to the revised official methods.
The features that distinguish this edition from the preceding
one are:
1. The contents of the book have been rearranged into three
parts instead of four. Part I treats of general methods of gravimetric and volumetric analysis, Part II treats of physicochemical methods, and Part III contains the special methods of
pharmaceutical analysis. In the deletion of Part IV of the first
edition pertaining to non-official methods, the chapters on
Hydrogen Ion Concentration, Photometric Methods, and
Electrolytic Methods have been placed in Part II under Physicochemical methods. The chapter on Ultimate Analysis has been
deleted to saye space and because a comprehensive treatment of
this subject properly belongs in a course in organic analysis.
2. The theory has been brought up to date and many of the
chapters have been largely rewritten. Sections on Calculations
and Errors, Significant Figures, and Logarithms and Their Use
have been added.
3. A number of new exercises have been added and many of
the former exercises have been replaced by new ones so that the
instructor has a wider range of selection.
4. The questions and problems have been extensively revised
and added to.
_ 5. The method of presentation employed in the first edition,
wherein typical analyses are explained simply, clearly, and
directly in considerable detail, has been retained. Extensive
tables classify all the official methods according to the type
methods which are described and explained.
vii
viii
PREFACE TO THE SECOND EDITION
The wide popularity that the first edition of thi~ book has
enjoyed among students and teachers has been gratifying to the
publishers and to the authors. It is hoped that the new edition
will similarly commend itself to a still larger circle. To the many
kind friends who have so generously aided by means of helpful
suggestions, the authors desire to express their thanks.
GLENN L. JENKINS.
ANDREW G. DuMEZ.
MINNEAPOLIS, MINN.
BALTIMORE, MD.,
February, 1937.
PREFACE TO THE FIRST EDITION

The object of this book is twofold: First, to furnish students of
pharmacy with a systematic course covering all of the quantitative chemical and physical methods official in the United States
Pharmacopoeia and the National Formulary through the selection and explanation of typical procedures. Second, to present
some of the generally applicable, non-official methods of analysis
which are widely used in pharmacy and with which all students
pursuing the profession should be familiar. In 'both instances,
the theory and practice of analytical chemistry as applied in
quantitative pharmaceutical procedures have been correlated.
The use of the book as a text should be supplemented by lecture and recitation instruction. It is obviously impracticable to
include detailed explanations of all of the quantitative determinations in the Pharmacopoeia and National Formulary. Typical
analyses illustrating all of the various methods have therefore
been selected and explained in considerable detail, while those
determinations requiring the same or closely similar procedures
are indicated in tables or otherwise. The instructor may select
other determinations from the Pharmacopoeia or National
Formulary and assign to the students the task of applying the
theory and explanation of the procedure involved as given in the
text.
The book is divided into four parts. Part I is intended for
use with students who have ~ompleted a full year of work in
general inorganic chemistry and qualitative analysis. Parts
II and III preferably should be given after the students have
eompleted inorganic and organic chemistry. Part IV should be
given to advanced students who have acquired a thorough knowledge of quantitative technique. In the authors' classes, Part I
is given during the second half of the second year in a course
covering sixteen weeks with one four-hour laboratory period
each week. In this course/ about twenty assays a.re performed
ix
in addition to the exercises pertaining to the analytical. balance

and standard solutions. Parts II and III are given in the second
half of the third year in a course of instruction. covering sixteen
weeks with two three-hour laboratory periods per week.
Approximately thirty-two exercises are performed during this
course other than those which deal with tl;te preparation of standard solutions, etc. Practically all of Part IV is taken up along
with other work in a course in food and drug analysis given in
the fourth year. The number of exercises that can be covered
in any given course will depend on the preparation of the students,
the amount of time devoted to the subject, and the facilities
available. The large number of exercises included provide for a
considerable degree of elasticity in the time devoted to the study
of quantitative methods and permit the instructor to vary the
exercises with different classes.
The authors wish to acknowledge their appreciation of the
courtesy extended by the following companies which granted
permission to use illustrations from their catalogues and books:
A. H. Thomas Company; Bausch and Lomb Optical Company;
Central Scientific Company; Christian Becker, Incorporated;
E. H. Sargent and Company; LaMotte Chemical' Products
Company; and Leeds & Northrup Company.
Notices of error and suggestions for the improvement of the
text will be greatly appreciated by the authors.
GLENN L. JENKINS.
ANDREW G. DuMEZ.
BALTIMORE,
MD.
December, 1930.
CONTENTS
PAGE
PREFACE TO THE SECOND EDITION.
vii
ix

INTRODUCTION .
xix
Definitions and scope of quantitative pharmaceutical chemistryReferences.

PART I
General Methods Used in Official Pharmaceutical Analyses
CHAPTER I
REMARKS AND GENERAL DIRECTIONS .
Sampling-Calculation of results and errors-General operationsThe analytical balance-Weights.

CHAPTER
GRAVIMETRIC ANALYSIS . . . . . .
II
29
Theory of ionization-Reversible reactions-Solubility product

principle-Common ion effect.
CHAPTER III
37
GRAVIMETRIC METHODS . .
Assay of sodium chloride, of sodium sulfate, of mercuric chloride, of

calcium glycerophosphate, of alum, and of solution of magnesium
citrate.
CHAPTER IV
PRINCIPLES OF VOLUMETRIC (TITRIM,ETRIC) ANALYSIS.
57
Definitions-Volumetric apparatus-The calibration of volumetric

apparatus-Neutralization .. method~: Theory-IndicatorsStandard solutions-Preparation and standardization of normal
hydrochloric acid, of normal sodium hydroxide, and of tenthnormal barium hydroxide.
CHAPTER V
ALKALIMETRY .
Direct titration methods: Assay of sodium bicarbonate, of sodium

hydroxide, and of sodium salicylate.
xi
86
xu
CONTENTS
PAGE
Residual titration methQds: Assay of zinc oxide, of potassium and

sodium tartrate, of magnesia magma, of methenamine; of solution of ammonium acetate, and estimation of nitrogen by' the
Kjeldahl method.
CHAPTER VI
ACIDIMETRY .
105
Direct titration methods: Assay of diluted sulfuric acid, of boric

acid, of tablets of sodium salicylate, and of tartaric acid.
Residual titration methods: Assay of aromatic sulfuric acid ~ntf of
tablets of acetylsalicylic acid.
CHAPTER VII
PRECIPITATION METHODS .
116
Determination of the end point-Indicators. Standard solktions.

Preparation and standardization of tenth-normal silver nitrate and
of tenth-normal ammonium thiocyanate.
Direct titration methods: Assay of strong silver protein.
Residual titration methods: Assay of sodium chloride, of ammonium bromide, of syrup of hydriodic acid, and of elixir of three
bromides.
V~II
CHAPTER
. . . . . . . . . . . .,129
Theory-Standard solutions-Preparation and stanaardization of
tenth-normal p'otassium permanganate.
Direct titration methods: Assay of ferrous sulfate, of reduced
iron, and of solution of hydrogen peroxide.
Indirect titration methods: Assay of calcium gluconate.
Residual titration methods: Preparation and standardization of
tenth-normal oxalic acid-Assay of sodium nitrite, of pre~
cipitated calcium carbonate, and of potassium chlorate.
Dichromate methods: Preparation of tenth-normal potassium
dichromate-Assay of mass of ferrous carbonate.
OXIDATION-REDUCTION METHODS .
CHAPTER IX
OXIDATION AND REDUCTION-IoDOMETRIC METHODS . . . . . . . . .
Starch indicator solutions. Standard solutions: Preparation and

standardization of tenth-normal sodium thiosulfate solution
and of tenth-normal iodine.
Direct titration with standard iodine solution: Assay of arsenic
trioxide.
Direct titration with sodium thiosulfate: Assay of compound
solution of iodine.
Residual titration with standard sodium thiosulfate: Assay of
mercurous chloride.
155
xiii
CONTENTS
PAGE
Titration of the iodine liberated from potassium iodide with sodium

thiosulfate: Assay of solution of ferric chloride, of chlorinated
lime, qf cupric sulfate, of sodium arsenate, of thyroid, and of
spirit of ethyl nitrite.
Titration with tenth-normal bromine: Preparation and standardization of tenth-normal bromine--Assay of phenol and of
ammonium hypophosphite.
Titrations with standard potassium iodate: Preparation of standard
potassium iodate solution-Assay of potassium iodide.
CHAPTER X
GASOMETRIC METHODS .
188
Theory...--Apparatus-Test of the nitrometer-Assay of carbon

dioxide and of spirit of ethyl nitrite.
PART II
Pl5.ysical Methods Used in Official Pharmaceutical Analyses
CHAPTER XI
SOLUBI}!iITY.
203
..
207
l)efinitions.
Determination of the solubility of boric acid in water at 25C.
CHAPTER XII
SPECIFIC GRAVITY AND DENSITY .
Methods used to determine the specific gravity of liquids: The use

of pycnometers-Determination of the alcohol content of an
official preparation. The use of the Westphal balance-Determination of the specific gravity of a volatile oil. The use of
hydrometers.
Methods used to determin~ the specific gravity of solids: B~ weighing in water-Determination of the specific gravity of camphor:
By the flotation method-Determination of the specific gravity
of yellow wax.
CHAPTER XIII
MELTING, CONGEALING, AND BOILING POINTS.
Melting point: Determination of the melting point of salicylic acid.

Congealing point: Method of determining-Determination of the
solidification temperature of the fatty acids of cottonseed oil.
Boiling and distilling point: Determination of the boiling point of
carbon tetrachloride.
226
xiv
CONTENTS
PAGE
CHAPTER XIV
REFRACTOMETRIC MEASUREMENTS .
. 240
Refractive index: Refractometers-The Abbe refractometer.

Determination of the refractive index of oil of oran~e.
CHAPTER XV
ROTATORY POWER .
247
Definitions-Polarimeters.
Determination of the specific rotation of sucrose.
CHAPTER XVI
. 257
VISCOSITY MEASUREMENTS.
Definitions-Apparatus: The Saybolt viscosimeter.

Determination of the kinematic viscosity of liquid petrolattlm.
CHAPTER XVII
PHOTOMETRIC METHODS OF ANALYSIS.
. .
. .
. .
. .
. .
260
Colorimetry: Determination of the ammonia contellt of waterDetermination of the amount of epinephrine hyclrochlori1.e in
solution of epinephrine hydrochloride-Assay of crPCUS for color.
Nephelometry: Determination of the amount of arsep-ic trioxide in
solution of arsenous acid-Determination of the aJjlount of oil of
peppermint in spirit of peppermint and limit test for chloride an.d
sulfate in calcium gluconate.
CHAPTER XVIII
DETERMINATION OF HYDROGEN ION CONCENTRATION
......
Acid base equilibrium and pH.

Potentiometric methods: The hydrogen electrode-rPlatinization
of the hydrogen electrode-The calomel electrode--The scheme
of assembly for hydrogen ion methods.
Determination of the end point of titration of hydrochloric acid
with sodium hydroxide potentiometrically-Notes and precautions-Determination of the end point of titro-tion of acetic
acid with sodium hydroxide potentiometrically. :J)etermination
of the pH of saturated boric acid solution, and or physiological
salt solution.
The quinhydrone electrode: Preparation-Use of.
Determination of the pH of elixir of iron, quinine aJld strychnine,
elixir of pepsin, and tincture of aconite by mean!! of the quinhydrone electrode.
The glass electrode.
Colorimetric methods: Indicators-Buffer solutions_,-Color standards-Color comparators.
Determination of the pH of solution of epinephrine P.ydrochloride,
and syrup of hydriodic acid.
272
CONTENTS
xv
CHAPTER XIX
ELECTROLYTIC ~ETHODS . . . . . . . . . . . . . . . . . . . . . 305
Electrical units and fundamentallaws-Theory-Apparatus.
Assay of copper sulfate and of mercuric chloride. Other electrolytic assays.
PART III
Special Methods
Use~
in Official Pharmaceutical Analyses
CHAPTER XX
ASH AND ~OISTURE DETERMINATIONS . . . . . . . . . . . . . . . 323
Ash content: Determination of the total and acid-insoluble ash
content of digitalis leaf.
~oisture content: Determination of the moisture content of acacia
-Determination of the moisture content of digitalis leaf by the
toluene distillation method.
CHAPTER XXI
ExTRACTIVE AND CRUDE FillER CONTENT . . . . . . . . . . . . . 336
Volatile and non-volatile ether-soluble extractive: Determination
of the volatile and non-volatile ether-soluble extractive of clove.
Alcohol-soluble extractive: Assay of benzoin.
Water-soluble extractive: Assay of aloe.
Purified petroleum benzin extractive.
Crude fiber: Determination of the crude fiber content of cloves.
CHAPTER XXII
CONSTANTS OF FATS, FATTY OILS, WAXES, BALSAMS, RESINS, ETC. . . 345
Acid number: Determination of the acid value of rosin.
Saponification value: Determination of the saponification value of
cottonseed oil.
Ester number.
Unsaponifiable matter.
Iodine value: Determination of the iodine value of olive oil.
CHAPTER XXIII
. . . . . . . . . . 361
ASSAY OF VOLATILE OILS
~ethods of general application: Specific gravity-Rotatory power
-Refractive index-Congealing point-Distilling point-Fractional distillation-Solubility.
Assay for ester content: Preparation of half-normal alcoholic potassium hydroxide-Assay of oil of peppermint for total esters.
Assay for alcohol content: Assay of oil of peppermint for total
menthol.
Assay for aldehyde content: Assay of oil of bitter almond for
benzaldehyde content.
xvi
CONTENTS
PAGE
Assay for ketone content: Assay of oil of caraway.

Assay for phenol content: Assay of oil of clove.
Assay for hydrocyanic acid content: Assay of oil of bitter almond
for hydrocyanic acid.
Assay 101: ascaridol content: Assay 01 oil of chenopodium.
Assay for allyl isothiocyanate: Assay of oil of mustard.
Assay for volatile oil in spirits: Assay of spirit of peppermint.
CHAPTER XXIV
ALKALOIDAL ASSAYING . . . . . . . . . . .
386
General principles: Sources of error-Theory of distribution

coefficient-Choice of indicators-Test solutions.
General procedures: Selection of the sample-Extraction with
immiscible solvents-Evaporation of organic solvents-Gravimetric determination of alkaloids-Volumetric determina'tion of
alkaloids.
CHAPTER XXV
OFFICIAL TYPE METHODS .
403
Alkaloidal assays by aliquot-part method.

General procedure: Extraction of the drug-Decanting the aliquot
portion-Shaking out with acid-Shaking out...with immiscible
solvent-Determination of the alkaloidal content.
Gravimetric assays: Assay of hydrastis for ether-solu\>le alkaloids, of cinchona for total alkaloids, and of compou.nd tincture of
cinchona.
Volumetric assays: Assay of ipecac for ether-soluble alkaloids and
assay of areca.
Alkaloidal assays by the total extraction method: Assay of hyoscyamus leaves.
Assay of preparations of hyoscyamus, belladonna, and stramonium:
Assay of tincture of belladonna and fluidextract of belladonna
leaf.
CHAPTER XXVI
ALKALOIDAL ASSAYS BY SPECIAL METHODS . . .
Assay of opium.
Table of official substances assayed by the same met.hod as opium.
Assay of colchicum.
Table of substances assayed by the same method as colchicum.
Assay of nux vomica.
Assay of caffeine containing drugs:-Assay of guaranlt.
Table of official drugs and preparations assayed for caffeine.
Assay of alkaloidal salts: Assay of citrated caffeille, of eucaine
hydrochloride, of theobromine with sodium salicylate, and of
theophylline with sodium acetate.
418
CONTENTS
~vii
PAGE
CHA'pTER XXVII
OTHER OFFICIAL ASSAYS INVOLVING THE USE OF IMMISCIBLE SOLVENTS 437
Assay of cantharides-Assay of aspidium-Assay of jalap-Assay
of tablets of phenobarbital.
CHAPTER XXVIII
ASSAY OF ENZYME-CONTAINING SUBSTANCES . . . . . . . . . . . . 444
Assay of pepsin-Assay of pancreatin for starch digestive powerAssay of pancreatin for casein digestive power-Asflay of rennin.
TABLE OF LOGARITHMS AND ANTILOGARITHMS.
453
INDEX . . . . . .
457
TABLE OF ATOMIC WEIGHTS . . . . . . . . . . . . lnside back cover
INTRODUCTION
Quantitative pharmaceutical analysis may be defined as the
application of the procedures of quantitative analytical chemistry
to the analysis of the materials used in pharmacy and, especially,
to the determination of the purity and quality of the drugs and
chemicals official in the United States Pharmacopoeia and in the
National Formulary. A complete chemical analysis of any
substance requires the establishment of the identity of its
component parts by qualitative analysis and the determination
of the proportion in which these components are present by the
processes of quantitative analysis.
Quantitative chemical methods are commonly classified as
organic and inorganic j gravimetric, volumetric, gasometric, and
electrometric. It is impractical, however, to adhere strictly
to this classification in presenting the subject matter of quantitative pharmaceutical analysis since certain phases of the work, as
alkaloidal assaying and the assay of essential oils, have been
developed to a point where' they constitute a distinct subject
matter in which the principles involved and the technique
required are the same whether the procedure be gravimetric or
volumetric.
Quantitative pharmaceutical analysis may be appropriately
subdivided according to various procedures each of which requires
a special technique, as follows:
Gravimetric analysis, or the separation, by extraction, precipitation, or other means of the constituent to be determined
either in the natural state, or in the form of a definite compound
the composition of which is known to the analyst, and weighing
the resulting product.
Volumetric analysis, or the determination of the volume of a
solution of known concentration required to react with a given
amount of the substance to be analyzed.
Gasometric analysis, or the measurement of the volume
of a liberated gas or the decrease in volume of a mixture of
xix
xx
INTRODUCTION
gases when a suitable reagent is used to remove one of the

gases present.
Physico-chemical methods of analysis, or those which are based
on the utilization of some specific physical or chemical property
or properties of the substance in its quantitative estimation.
Those physico-chemical methods commonly applied in quantitative pharmaceutical chemistry are optometric, refractometric,
electrometric, and colorimetric. They comprise some of the
most valuable methods used in analytical procedure. Since
each of them require special treatment, they will be discussed
separately.
Special methods, or those which require a distinct type of
technique, as alkaloidal assaying, require separate treatment in
order to preserve unity of subject matter.
The official assay methods serve as an exact measure of the
purity of a substance only when the results are considered in
conjunction with the qualitative tests. Thus, in the assay of
zinc oxide, the purity of the oxide as determined by assay must
follow qualitative tests for other metals which if present would
be estimated as zinc oxide. A complete analysis, therefore,
requires the qualitative identification of all substances present
as well as their quantitative estimation.
The theoretical considerations applicable in any given analysis
are inherently a part of the analytical procedure. They should
be mastered before a determination is made and carefully applied
throughout each step of the procedure.
Those who desire a more comprehensive treatment of special
aspects of analytical procedure will find the following texts and
reference books of value:
Textbooks
1. BASSET, "The Theory of Quantitative Analysis," Alfred A. Knopf, Inc.,'
New York, 1925.
2. BLASDALE, "Principles of Quantitative Analysis," 3d ed., D. Van
Nostrand Company, Inc., New York, 1928.
3. CLOWES and COLEMAN, "Quantitative Chemical Analysis," 13th ed.,
P. Blakiston's Son & Company, Philadelphia, 1931.
4. CUMMINS and KAY, "A Textbook of Quantitative Chemical Analysis,"
6th ed., Gurney and Jackson, London, 1934.
5. ENGELDER, "Elementary Quantitative Analysis," 2d ed., John Wiley
& Sons, Inc., New York, 1936.
INTRODUCTION
XXI
6. FALES, "Inorganic Quantitative Analysis," D. Appleton-Century

Company, Inc., New York, 1925.
7. FARNSWORTH, "Theory and Technique of Quantitative Analysis,"
John Wiley & Sons, Inc., New York, 1928.
8. FAY, "An Advanced Course in Quantitative Analysis," 2d ed., John
Wiley & Sons, Inc., New York, 1922.
9. GUY and SKEEN, "A Course in Quantitative Analysis," Ginn and
Company, Boston, 1932.
10. HALL, "A Textbook of Quantitative Analysis," 2d I'd., John Wiley &
Sons, Inc., New York, 1935.
11. HENDEL, "Quantitative Analysis," Ginn and Company, Boston, 1925.
12. KOLTHOFF and SANDELL, "Textbook of Quantitative Inorganic Analysis," The Macmillan Company, New York, 1936.
13. MAHIN, "Quantitative Analysis," 4th I'd., McGraw-Hill Book Company, Inc., New York, 1932.
14. OLSEN, "Quantitative Analysis," 5th I'd., D. Van Nostrand Company,
Inc., New York, 1919.
15. POPOFF, "Quantitative Analysis," 3d I'd., P. Blakiston's Son & Company, Philadelphia, 1935.
16. RIEMAN and NEUSS, "Theoretical Approach to Quantitative Analysis,"
McGraw-Hill Book Company, Inc., New York, 1937.
17. SCHIMPF, "Essentials of Volumetric Analysis," 4th ed., John Wiley &
18. SCOTT, "Essentials of Quantitative Chemical Analysis," 2d cd., Chemical Publishing Company, Easton, Pa., 1931.
19. SMITH, "Quantitative Chemical Analysis for Beginning Students,"
3d cd., The Macmillan Company, New York, 1933.
20. STOCK and STAHLER, "Quantitative Chemical Analysis," McGrawHill Book Company, Inc., New York, 1935.
21. TALBOT, "Introductory Course in Quantitative Analysis," 7th ed., The
Macmillan Company, New York, 1931.
22. WILLARD and FURMAN, "Elementary Quantitative Analysis," 2d ed.,
D. Van Nostrand Company, Inc., New York, 1936.
Calculations
1. HAMILTON and SIMPSON, "Calculations of Quantitative Analysis,"
2d ed., McGraw-Hill Book Company, Inc., New York, 1927.
2. LONG and ANDERSON, "Chemical Calculations," 3d I'd., McGraw-Hill
Book Company, Inc., New York, 1936.
3. MELLOR, "Higher Mathematics for Students of Chemistry and
Physics," 4th I'd., Longmans, Green & Company, New York, 1922.
4. MILLER, "Calculations of Analytical Chemistry," 3d I'd., McGrawHill Book Company, Inc., 1921.
5. MOORE, "Logarithmic Reduction Tables, for Students of AnaJytical
Chemistry," Ginn and Company, Boston, 1913.
xxii
INTRODUCTION
6. WILKINSON, "Calculations in Quantitative Analysj.s," McGraw-Hill

General References
l. ALLEN, "Commercial Organic Analysis," 5th ed., P. Blakiston's Son &
Company, Philadelphia, 1926.

2. GOOCH, "Methods in Chemical Analysis," John Wiley & Sons, Inc.,
New York, 1912.
3. GRIFFIN, "Technical Methods of Analysis," McGraw-Hill Book Company, Inc., New York, 1921.
4. KOLTHOFF and FURMAN, "Volumetric Analysis," JollD Wiley & Sons,
5. MELLOR, "A Treatise on Quantitative Inorganic Analysis," Charles
Griffin & Company, Ltd., London, 1913.
6. SCOTT, "Standard Methods of Chemical Analysis," 4th ed., D. Van
7. SMITH, "Analytical Processes," Edward Arnold and do., London, 1929.
8. SUTTON, "Volumetric Analysis," 12th ed., P. Blakiston's Son & Company, Philadelphia, 1935.
9. TRE.A.DWE.L.L. and HA.LL., "Q.uantitative A.n.al,!si"':' stU ed ... John Wiley
& Sons, Inc., New York, 1935.
10. VILLAVECCHIA, "Applied Analytical Chemistry," p.J3lakiston's Son &
Drug Analysis
1. DRAGENDORFF,
2.
3.
4.
5.
6.
7.
8.
"Plant Analysis, Qualitative and Quantitative,"

Balliere, Tindall and Cox, London, 1884.
EVERS and ELSDON, "Analysis of Drugs and Chemicals," Charles
Griffin and Company, London, 1929.
FULLER, "The Chemistry and Analysis of Drugs and Medicines," John
LYONS, "Practical Standardization of Drugs," Nelson & Co., Detroit,
1920.
"National Formulary VI," 6th ed., Mack Printing Oompany, Easton,
Pa., 1936.
NELSON, "Introduction to the Analysis of Drugs and Medicines," John
"New and Non-official Remedies" ("N.N.R."), MDerican Medical
Association, Chicago, 1936.
"Pharmacopoeia of the United States XI," 11th rev., Mack Printing
Company, Easton, Pa., 1936.
Food Analysis
1. BLYTH, "Foods, Their Composition and Analysis" 6th ed., D. Van

INTRODUCTION
xxiii
2. BRIDGES, "Food and Beverage Analysis," Lea Febeger, Philadelphia,

1935.
3. Cox, "Chemical Analysis of Foods," P. Blakiston's Son & Company,
Philadelphia, 1926.
4. LEACH and WINTON, "Food Inspection and Analysis," 4th ed., John
Wiley & Sons., Inc., New York, 1920.
5. LEFFMAN and BEAM, "Food Analysis," P. Blakiston's Son & Company,
Philadelphia, 1905.
6. SHERMAN, "Organic Analysis," The Macmillan Company, New York,
1920.
7. WOODMAN, "Food Analysis," 3d ed., McGraw-Hill Book Company,
Food and Drug Analysis
1. LIVERSEEGE, "Adulteration and Analysis of Foods and Drugs," D.

Van Nostrand Company, Inc., New York, 1932.
2. MOOR and PARTRIDGE, "Aids to, the Analysis of Food and Drugs,"
5th ed., William Wood and Company, Baltimore, 1935.
3. "Methods of Analysis or'the Association of Official Agricultural Chemists" (A.O.A.C.), 3d ed., Assoc. Off. Agr. Chem., Washington, D. C.,
1932.
\
4. PARRY, "The Analysis of Food and Drugs, Chemical and Microscopical,"
Scott, Greenwood & Sons, London, 1911.
5. THURSTON, "Pharmaceutical and Food Analysis," D. Van Nostrand
Special Analytical Methods
1. BLAIR, "The Chemical Analysis of Iron," 8th ed., J. B. Lippincott
2. BRITTON, "Conductimetric Analysis," D. Van Nostrand Company,
3. - - - , "Hydrogen Ions. Their Determination and Importance in
Pure and Industrial Chemistry," D. Van Nostrand Company, Inc.,
New York, 1932.
4. CLARK, "Determination of Hydrogen Ions," 3d ed., Williams & Wilkins
Company, Baltimore, 1928.
5. DENNIS, "Gas Analysis," The Macmillan Company, New York, 1913.
6. DIETERICH, "Analysis of Resins," Scott, Greenwood & Sons, London,
1920.
7. -GILDEMEISTER, HOFFMAN, and KREMERS, "The Volatile Oils," John
8. HALDANE and GRAHAM, "Methods of Air Analysis," 4th ed., Charles
Griffin & Company, Ltd., London, 1935.
9. HEVESY, "Chemical Analysis by X Rays and Its Application," Cornell
University Press, Ithaca, N. Y., 1932.
xxiv
I NTRODUCT TON
10. HILLEBRAND and LUNDELL, "Applied Inorganic Analysis with Special

Reference to the Analysis of Metals, Minerals, and Rocks," John
11. KOLTHOFF, "The Colorimetric and Potentiometric Determination of
pH," John Wiley & Sons, Inc., New York, 1931.
12. - - - , "Conductimetric Titrations," T. Steinkopff, Leipzig, 1923.
13. - - - , and FURMAN, "Potentiometric Titrations," 2d ed., John
14. LACEY, "Instrumental Methods of Chemical Analysis," The Macmillan
Company, New York, 1924.
15. LEWKOWITSCH, "The Chemical Technology and Analysis of Oils, Fats
and Waxes," 6th ed., The Macmillan Company, New York, 1923.
16. LUNGE, "Technical Gas Analysis," D. Van Nostrand Company, Inc.,
New York, 1934.
17. MAHIN and CARR, "Quantitative Agricultural Analysis," McGraw-Hill
18. PETERS and VAN SLYKE, "Quantitative Clinical Chemistry," Williams
& Wilkins Company, Baltimore, 1932.
19. PREGL, "Quantitative Organic Microanalysis," 2d English ed., P.
Blakiston's Son & Company, Philadelphia, 1930.
20. RADLEY and GRANT, "Fluorescence Analysis in Ultraviolet Light,"
D. Van Nostrand Company, Inc., New York; 1933.
21. RAE and REILLY, "Physico Chemical Practical Exercises," Methuen

and Company, Ltd., London, 1935.
I
22. SMITH, "Electro-analysis," 6th ed., P. Blakiston's Son & Company,
Philadelphia, 1918.
23. SNELL and SNELL, "Colorimetric Methods of Analysis, Including Some
Turbidimetric and Nephelometric Methods," D. Van Nostrand
24. WINKLER, "Handbook of Technical Gas Analysis," 2d English ed.,
Gurney and Jackson, London, 1932.
25. WRIGHT, "Soil Analysis," Thomas Murby and Company, London,
1934.
26. YOE, "Photometric Chemical Analysis, Colorimetry," John Wiley &
27. - - - , "Nephelometry," John Wiley & Sons, Inc., New York, 1929.
References on Physical and Chemical Data
1. ATACK, "The Chemists Year Book," Chemical Catalog Company, Inc.,
New York, 1936.

2. COMEY and HAHN, "A Dictionary of Chemical Solubilities, Inorganic,"
The Macmillan Company, New York, 1921.
3. HODGMAN and COOLBAUGH, "Handbook of Chemistry and Physics,"
The Chemical Rubber Co., Cleveland, 1936.
4. "International Critical Tables," McGraw-Hill Book Company, Inc.,
New York, 1926.
INTRODUCTION
xxv
5. LANDOLT-BoRNSTEIN,. "Physicalisch-Chemische Tabellen," Julius

Springer, Berlin, 1923.
6. MURRAY, "Standards and Tests for Reagent Chemicals," D. Van
7. OLSEN, "Van Nostrand's Chemical Annual," D. Van Nostrand Company, Inc., New York, 1928.
8. SEIDELL, "Solubilities of Inorganic and Organic Chemicals," D. Van
Nostrand Company, New York, 1928.
9. "Smithsonian Physical Tables," The Smithsonian Institution, Washington, D. C., 1918.
PART I
GENERAL METHODS USED IN OFFICIAL
PHARMACEUTICAL ANALYSES
The theoretical considerations and the procedures involved in

ravimetric, volumetric, and gasometric methods of analysis
re the same in quantitative pharmaceutical analysis as in quanitative chemical analysis. In quantitative pharmaceutical
,nalysis, however, the general theories of analytical chemistry
,re applied to the procedures used in the analysis of the drugs,
hemicals and medicinal preparations employed in pharmacy;
:hiefly to those materials official in the United States Pharma:opoeia(U.S.P.) and National Formulary (N.F.).
CHAPTER I
REMARKS AND GENERAL DIRECTIONS
Success as an Analyst.-To be successful as an analyst, the
student must realize that analytical chemistry is not simple
routine procedure. Manipulative skill acquired by experience
with the ability to follow directions under the supervision of a
skilled analyst may enable one to carry out successfully certain
analytical procedures. The object of teaching quantitative
pharmaceutical chemistry, however, is to impart a thorough
fundamental knowledge of the theory as well as the practice of analytical methods.
The successful analyst must know what reactions are taking
place during an analysis and be able to understand and apply
the theory upon which the method is dependent. The analyst
must acquire skill of technique, patience, neatness, and accuracy.
The fact should be borne in mind that analysis is not carried out
for the sake of analysis but as a means to control the purity and
strength of substances used as medicaments. Above all, to be
successful, the analyst must not only use common sense in the
laboratory but think throughout each step of the procedure.
Accuracy and Honesty.-It is absolutely essential that all
quantitative work be performed without even a slight loss of
material or gain of extraneous matter. Every precaution should
be taken to prevent such loss or gain. All solutions and precipitates should be covered to prbtect them from dust, whenever
possible, and every particle of sample, solution or precipitate
must be regarded as indispensable to the ultimate success of the
analysis.
Absolute integrity is demanded of every quantitative analyst.
The operator is the only person who is familiar with the entire
history of an analysis and who knows whether it is worthy of full
confidence. When confidence is lost for any definitely established reason in the course of an analysis, operations should be
3
QUANTITATIVE PHARMACEUTICAL CHEMISTRY
discontinued at once and a new start made. All determinations

should be made in duplicate, and the results should agree closely.
Failure to obtain results which check is conclusive evidence that
at least one result is wrong and that neither can be depended upon.
Notebooks.-All notebooks should be of the type designated
by the instru0tor. A small bound notebook which will slide
under the balance while weighing has proved most satisfactory
in the writer's experience.
The following information relative to each determination
should be included:
Date.
Object or title.
Experimental data.
Reactions.
Calculations.
Results.
Remarks.
This information should be arranged In a neat, systematic

manner so that anyone familiar with quantitative analysis may
understand it readily.
The date is very important in all scientific work. Numerous
legal decisions have been lost through the failure of the analyst
to affix the date at the time of analysis. The object or title
should be briefly stated. The experimental data should include
a complete record of all weighings and measurements set down
at the time they are made. The practice of recording data on
loose pieces of paper or in a scrap notebook and copying the
original entries into a second notebook should not be tolerated.
These data may be conveniently placed and followed by the calculations on the left-hand page of the notebook, leaving the
right-hand page for a concise, orderly write-up of the entire
experiment. All reactions should be given using structural
formulae where possible. The use of logarithms for all calculations is highly recommended for accuracy and economy of time.
Results should always be reported as percentages unless otherwise directed. Remarks should include any unexpected development during the analysis, explanation of error, and comments
which seem pertinent to the technique or theory involved in
the analytical method.
Economy of Time.-Economical utilization of laboratory

hours is best achieved through a preliminary study of the work
to be done, followed by a plan for its prompt execution. There
is no such thing as "speed" in quantitative work. Rapidity
of accomplishment is achieved not by haste but by planning the
work so that two or more operations may be carried out at one
time and especially by being so accurate that it will be unnecessary to repeat a determination. The following suggestions will
save time and increase accuracy: (1)
Keep the desk scrupulously clean at
all times. (2) Clean all apparatus at
once when through using it for the
day and put it in the desk. (3)
Label all solutions, filtrates, and
precipitates systematically throughout the analytical procedure. (4)
Keep two or more operations going
at one time; thus, while cooling a
precipitate in the desiccator, make a
weighing, or while washing one
precipitate, ignite another one. (5)
Utilize all time between operations
in making calculations and writing
up experiments.
Cleaning Mixture.-A good clean- FIG. l.-Wash bottle for distilled water.
ing mixture may be prepared by
adding approximately 40 Gm. of commercial sodium dichromate to 400 Gm. of commercial, concentrated sulfl~ric acid and
stirring the mixture. Commercial potassium dichromate may be
used, but the sodium dichromate is cheaper and more soluble in
concentrated sulfuric acid.
This mixture should be used to wash all glass and porcelain
ware. The mixture is best preserved in a thick-walled, glassstoppered, 500 cc. bottle.
Wash Bottles.-Wash bottles for use in quantitative work
usually consist of rubber-stoppered, flat-bottomed flasks of 500
to 1,000 cc. capacity, fitted with smoothly bent tubes and a jet
made flexible by means of a rubber joint. The jet should be
drawn out so that it will deliver a thin, even stream of liquid.
(3
Prepare two wash bottles of the type illustrated in Fig. 1, one of

1,000 cc. and one of 500 cc. capacity, for cold and hot water.
The necks of wash bottles for use with hot water should be
wrapped with asbestos paper or twine or other suitable
material.
To bore a hole through a rubber stopper, dip the end of the
borer in a strong solution of sodium hydroxide before starting to
bore. Boil all rubber parts in dilute sodium hydroxide for from
2 to 3 min. and wash them thoroughly with distilled water. Fire
polish the ends of all glass tubing by holding them in the blue
flame of a burner until the sharp edges have become rounded.
"Policeman."-The so-called "policeman" is a piece of soft
rubber tubing fitted to the end of a glass rod. The rod should be
about 20 cm. long with a diameter between 5 and 7 mm., and it
should have plain annealed ends. A piece of rubber tubing
about 3 cm. long should be fitted tightly over and project about
2 cm. beyond the rod. The policeman is used to loosen from
the walls of the containing vessel adhering particles which are
not removable by a stream of water ftom the wash bottle. A
policeman should never be used as a stirring Irod or allowed to
remain in analytical solutions.
Reagents.-The reagents used in quantitative analysis must
be pure. Reagents which conform to the requirements of the
United States Pharmacopoeia and of the National Formulary
should be used in all assays which have for their purpose the
establishment of the strength or quality of official substances.
Those reagents which meet the specifications of the American
Chemical Society as well as those sold by reputable manufacturers
as "analysed" or "reagent" grade will usually be found to meet
all official requirements.
Purity and Strength Requirements.-The purity and strength
of chemicals and drugs of the U.S.P. and of the N.F. are usually
expressed in terms of per cent. The official requirements of
purity and strength obtain only when the official methods of
assay are employed. When the per cent strength or purity is
given in round numbers, it is understood that the figures represent exactly the minimum or maximum requirement, e.g., the
requirement that hydrochloric acid contain "not less than 35 per
cen~ and not more than 37 per cent of HCI" permits a product Of
35 or 37 per cent or any intermediate strength; in other words, the

figures are inclusive. It is established by rule that the figures
35 and 37, respectively, have the same significance as the figures
35.00 and 37.00.
Materials Required.-The statement of materials required
in the exercises is intended as a guide for the instructor in ordering
supplies. Commonly used reagents and solutions such as indicators, acids, and alkalies are not always listed since it is assumed
that they are available in the analytical laboratory. The quantities indicated are for each sample. It is possible to give only
approximate quantities for materials used as solvents, for washing,
to make alkaline, and so forth. All reagent materials should
meet the official requirements of purity and strength.
SAMPLING
The first important consideration in an analysis is the securing

of a representative sample, a consideration too frequently
ignored by analysts. Fixed rules cannot be laid down in sampling for much depends on the nature of the material and the
quantity from which a representative sample is to be taken.
Whenever possible, a sample from each package should be examined separately. When a single package is sampled, the contents
should be mixed or samples should be taken from different parts
of the container and mixed, and then a portion of this mixture
should be taken for analysis. The sampling of bulk quantities
of chemicals and drugs may be performed by the methods given
in the U.S.P. for sampling vegetable drugs:
"1. It is recommended that gross samples of vegetable drugs
in which the component parts are 1 cm. or less in any dimension,
and all powdered or ground drugs, be taken by means of a sampler
which removes a core from the top to the bottom of the container,
not less than two cores being taken in opposite directions; that
when the total weight of the drug to be sampled is less than
100 kilos (200 pounds) at least 250 Gm. shall constitute an official
sample; that when the total weight of the drug to be sampled is in
excess of 100 kilos, repeated samples shall be taken by the above
method, and according to the schedule given below, mixed and
quartered, two of the diagonal quarters being rejected, the
remaining two quarters being combined and carefully mixed, and

again subjected 0 a quartering process in the same manner until
two of the quarters weigh at least 250 Gm., which latter quarters
shall constitute an official sample.
"II. It is recommended that gross samples of vegetable drugs
in which the component parts are over 1 cm. in any dimension
be taken by hand; that when the total weight of the drug to be
sampled is less than 100 kilos, at least 500 Gm. shall constitute
an official sample, and this shall be taken from different parts of
the container or containers; that when the total weight of the
drug to be sampled is in excess of 100 kilos, repeated samples
shall be taken by the above method and according'io the schedule
below, mixed and quartered, two of the diagonal quarters being
rejected, and the remaining two quarters being combined and
carefully mixed, and again subjected to a quartering process
in the same manner until two of the quarters weigh not less than
500 Gm., which latter quarters shall constitute an official sample.
SCHEDULE RECOMMENDED FOR SAMPLING
Number of Packages
in Shipment
1 to 10
10 to 25
25to 50
50 to 75
75 to 100
Number of Packages
to Be Sampled
1 to 3
2 to 4
3to 6
6 to 8
8 to 10
"When over 100, the total number sampled should not be

less than 10.
"III. When the total weight of a drug to be sampled is less
than 10 kilos, it is recommended that the above methods be
followed but that somewhat smaller quantities be withdrawn, and
in no case should the final official sample weigh less than 125 Gm.
I "IV. In additiori to the withdrawing of official samples according to methods I, II, and III, the official sample may consist of
the total amount of a direct purchase made by Federal, State or
Municipal Food and Drugs' Act enforcement officials."
Students' samples are supplied ready for analysis as a rule.
Samples which require drying should be spread on a large watch
glass, thoroughly mixed, and dried in an oven at the specified
temperature. They should then be transferred to a weighing

bottle and kept in the desiccator.
CALCULATION OF RESULTS AND ERRORS
Source and Nature of Errors.-It is only in rare cases that the

numerical value of an experimental result can be directly determined. As a rule, it is necessary to calculate the result from the
different measurements and observations which have been made.
Duplicate results which check very closely are not an assurance
of accuracy, as is often taken for granted by the beginner in
quantitative work, since errors inherent in the method and
apparatus used may be repeated for each sample. Generally,
results which agree closely when obtained by two different methods of analysis are a good indication of the reliability of the
methods.
The results of repeated analyses or measurements will fail to
agree, in general, when made by the same or different analysts
to the full precision of which the method or instrument is capable.
The discrepancies in the results are caused by various sources of
error to which all experimental data are subject. The errors
are of two types, namely: indeterminate or accidental errors
which manifest themselves by slight variations in a series of
observations made by the same observer under identical conditions. They result from causes difficult to detect, such as differences in the judgment and skill of the analyst. Generally,
indeterminate errors are intangible and their elimination by the
analyst is impossible. Determinate or constant errors are of
such nature that they recur in a constant manner'in each of a
series of determinations. Consequently, it is possible partially to
determine their value and reduce their effect on the final result.
They arise from causes such as: (1) Personal errors made by the
individual analyst, e.g., inability to judge color changes sharply,
resulting in habitual reading of end points in titration too late.
(2) Errors of method caused by faulty procedure, e.g., incorrect
sampling, contamination of precipitates, and improper selection
of indicators. (3) Apparatus errors due to poor construction or
calibration, e.g., inaccuracy in the calibration of burettes or
pipettes, inequality in the length of the arms of the balance, and
10
incorrect weights. Errors of this type are usually detectable

and so may be eliminated to a large extent.
Indeterminate Errors.-Indeterminate errors follow the law
of' chance which is represented by the following curve commonly
known as the curve of error or the probability curve.
An examination of this curve shows that: (1) Very large errors
are unlikely to occur often; (2) small errors occur with greater
frequency than large ones; and (3) positive and negative errors
of the same numerical magnitude are equally probable since the
curve is symmetrical with respect to the Y axis. Consequently
y
M"gnitude of error
FIG. 2.-Probability curve.
+X
it is apparent that the best value to select for a series of observations is their arithmetical mean.
'
Average Deviation from the Arithmetical Mean.-In order to
measure the magnitude of indeterminate or accidental errors,
use is made of the theorem proved by the method of least squares
which states that an arithmetical mean computed from n equally
trustworthy observations is
times as trustworthy as any
single observation. If the indeterminate error in anyone observation is represented 'by i, the indeterminate error of the arithmetical mean is i/Vn; in other words, the uncertainty of the
arithmetical mean is inversely proportional to the square root
of the number of observations. The deviation of a single measurement from the arith~etical mean of a series of similar measurements can readily be determined and from this the average
deviation of a single determination may be calculated. For
example: (1) In the standardization of hydrochloric acid by
titration against pure sodium carbonate, the burette readings
30.25, 30.22, 30.26, 30.27, and 30.23 cc. were obtained. The
vn
R'EMARKS AND GENERAL DIRECTIONS
11
arithmetical mean of these numbers is 30.246. The deviations

of the individual readings from the mean, neglecting sign, are
0.004, 0.026, 0.014, 0.024, and 0.016. The sum of these five
numbers is 0.084, and the average is 0.084/5 = 0.017. Expressing the average deviation as a ratio to the arithmetical mean
gives 0.017 part per 30.246 or about 0.56 part per 1,000 parts.
(2) Samples of a pure chloride weighing 0.5056 Gm. yielded calculated results in grams of 0.4850, 0.4840, 0.4844, 0.4846, and
0.4887, when the chloride ion was precipitated as silver chloride.
Calculate the inaccuracy (absolute error) in per cent and the
mean error in parts per thousand from the results. The mean
value obtained by experiment and calculation, rejecting the last
determination (see Rejection of a Result), equals 0.4845. The
absolute error is 0.5056 - 0.4845 = 0.0211. The relative error
of the method is 0.0211/0.5056 X 100 = 4 per cent (approximate). The mean error of the result is 0.06 per cent or 0.6 part
per 1,000 parts.
The example shows that the inaccuracy of the method used is
40 parts per 1,000 parts as compared with a mean error in the
determinations of 0.6 part per 1,000 parts.
Rejection of a Result.-Sometimes in a series of determinations, one of the results obtained will differ greatly from the
others without any apparent mistake having been made in the
work. The question then arises as to whether this result should
be retained with the others in calculating the value of the arithmetical mean. If at least four determinations have been made,
the result in which the error appears to be large may be omitted
and the arithmetical mean and the average deviation of the
other results computed. If the difference between the arithmetical mean and the result believed to be in error is four or
more times the average deviation, the result should be rejected,
e.g., in the preceding .example, the last result should be rejected
since 0.4887 - 0.4845 = 0.0042 and this difference is more than
four times the average deviation, 0.0003. It has been shown that
the deviation of a result which is four or more times the average
deviation is due to a mistake in all except about five cases in ~
thousand.
Significant Figures.-8ignificant figures are the digits which
when placed in order give the value to a number, e.g., in the
12
quantity represented by the number 236, the digits 2, 3, and 6

are significant figures. Zeros are employed in some cases to
locate the decimal point; in other cases, they may be significaI).t
figures. Thus in the number 20.5, the zero is a significant figure,
but in the number 0.0125 the zeros are not significant but show
the order of magnitude of the other digits. In the quantity,
1.0000 Gm., the zeros are significant since they indicate that the
weighing 'has been made to Ylo mg. and that only the last zero
is uncertain.
Every measurement involves some error or can be carried out
only with a certain degree of accuracy. Consequently, it is
evident that the number expressing the value of a measurement
can be only approximate. Irrespective of the number of figures
written as the result of a calculation, the accuracy of a value
beyond that limited by the errors of the measurements cannot
be increased. On the other hand, if too few figures are written
down, the resulting value may be much less accurate than the
data permit. The proper manner of expressing a result is to
retain or use such a number of figures that all, except the last,
are known with certainty, and that while the last figure is uncertain the error is not greater than 5 in the following place.
For example: If a burette can be read to 0.01 cc. and 32.5 instead
of 32.50 is written as the result of a measurement, the mistake of
writing too few figures is committed since the number 32.5
indicates that the true value lies between 32.45 and 32.55 and
the apparent error is five times greater than the error of measurement. Again if, in recording the result of a series of burette
readings, the mean of the values 32.52, 32.54, 32.54, and 32.55 cc.
is written as 32.538, it would be a mistake of retaining too many
figures since it would indicate that the error of measurement is
only about 0.0005 cc. The number should be rounded off,
therefore, to 32.54 cc. For purposes of further calculation, all
of the figures should be retained, i.e., the number 32.538 should
be used.
In general, for analytical work, the operations of addition and
subtraction should give as the final result no more decimal
places than the least number of decimal places entering into
the calculation. This is illustrated by the following example:
In the addition 12.4 + 121.502 + 3.6653 = 137.5673, the maxi-
13
mum apparent error in the result is 0.05 which is the greatest apparent error in the number 12.4. Therefore, a derived
error of 5 exists in the second decimal place, figure 6, of the
result and the1r"esult should be written 137.6 but if the result is
used for further calculations, it is better to retain one figure more
i.e., to employ 137.57.
In multiplication or division, as a rule, retain one more significant figure, when available, in the result than there are in the
number having the least number of significant figures. For
example: The product of 11.32 X 12.2 X 0.0321 = 4.4331384.
If the last figure in each number is uncertain, the relative error
in the number 12.2 will be about 5 in 1,200 or 0.4 per cent.
Consequently, it would be incorrect to write the result as
4.4331384 for this result has a derived error of 0.4 per cent or
of about 2; units in the second place of the decimals. All figures
following this are therefore meaningless and should be discarded,
the result. being written 4.43.
Logarithms and Their Use.-The solution of problems and
the calculation of the results of analyses from data are greatly
facilitatEld by the use of logarithms. It is advisable for students
to make calculations by the use of logarithms in order to save
time and avoid tedious calculations by long hand.
A lOgarithm is an exponent which must be applied to a fixed
positive number other than 0 or 1 to produce any given number.
The fixed number is called the base. In the common or Briggsian
system of logarithms, the base is 10. A logarithm consists of
twO' parts, the characteristic or integral part and the mantissa or
fractional part. The characteristic of a number greater than 1 is
always one less than the number of digits to the left of the decimal
point, i.e., where n equals the number of digits to the left of the
decimal point, the characteristic is represented by a number
equal to n - 1. For exampl~, the characteristic of 10,000 or
10 4 is 4, of 100 or 10 2 is 2, and of 1 or 10 0 is 0. The characteristic
may be either positive or negative. When there are no digits
to the left of the decimal point, the characteristic is negative or
minus, and when there are no zeros before the significant figures
to the right of the decimal point, the characteristic is -lor
9 - 10. When there are no digits to the left of the decimal
point and n zeros to the right of the decimal point before the
14
QUANTITATIVE PHARMACEUTICAL qHEMISTRY
significant figures, the characteristic is (9 - n) - 10, e.g., the

characteristic of 0.1 is I or 9 - 10, of 0.01 is 2,or 8 - 10, and
of 0.000001 is 6 or 4 - 10.
The mantissas of numbers are furnished by the table of logarithms, see page 453. The examples given below illustrate
the use of the tables:
Example I.-Find the logarithm of 9465. From the explanation given, it is evident that the characteristic is 3. To find the
mantissa, locate 94 in the logarithm table in the column of
Natural Numbers and then move directly to the right to the
column under 6 at the top of the page. The mantissa given there
is 9759. This is the mantissa for 9460. To find the mantissa
for 9465, move farther to the right to the coluJPns headed
Proportional Parts where, under the 5 column, the, figure 2 is
given. This figure 0002 when added to 9759 shows 'flow much
more the logarithm of 9465 is than that of 9460. Con~equently,
the logarithm of 9465 is 3.9759 + 0.0002 = 3.9761.
Example 2.-Find the logarithm of 0.0001058. Since the first
significant digit is four places to the right of the decimal point, the
characteristic is 6 - 10. Find the mantissa as in example 1 by
following the horizontal line opposite 10 to the vertical ,column
headed by 5 and add to this the proportional part un&er the
column headed 8. The mantissa is then found to be 0212 +
0033 = 0245 and the logarithm is written 6.0245 - ,10.
Example 3.-Multiplying and dividing with logarithms.
To multiply, add the logarithms of the numbers together. To
divide subtract one logarithm from the other. Study the folk)'wing example:
24.46 X 0.2917
1000 X 0.003741
Multiply by adding the logarithms
'log 24.46. . . . . . . . . . . . . . . . . . . . . .
1.3885
log 0.2917 ..................... = 9.4728 - 10
24.46 X 0.2917 is the Bum of the
logs ........................ = 10.8613 - 10
REMAfiKS AND GENERAL DIRECTIONS
3.0000
7.5730 - 10
log 1000.,. .................... .

log 0.00~41 .................. .
1000 X10.003741 is the sum of
the logs .....................
15
10.5730 - 10
Divide by subt);acting the logs

10.8613 - 10
10.5730 - 10
0.2883
The result of these operations is the logarithm of the number
sought. T,b find the natural number corresponding to the
logarithm ,0.2883, turn to the table of antilogarithms (page 455),
move down the column headed Logarithms until 28 is found,
move horizontally to the column headed 8 where the number 1941
is found" and then proceed to the Proportional Parts column
under 3 where the number 0001 is found; add this to 1941, the
sum is 1942. The characteristic is O. Therefore, one digit is
to the left of the decimal point and the required number is 1.942.
Questions and Problems
1. Rewrite each of the following quantities, underscoring the significant
figures: (a) 1.1200 Gm. weighed on the analytical balance, (b) 20.459 cc.
from a burette reading, (c) 0.0062 Gm. from a weighing on the analytical
balance.
2~ Make the following calculations retaining only the significant figures:
a. 2.3 + 100.56 + 4.273 - 20.005 =
b. 100 X 0.010 X 1000 + 10 =
c. (20 X 1.0250) - (10 X 1.2000) + 1.200 =
3. A sample known to be 98.50 per cent pure yielded results on analysis
in duplicate of 98.30 and 98.12 per cent, respectively. What is the precision
in parts per thousand of each determination?
4. The result of an analysis for per cent purity employing the usual
apparatus and weighing to the fourth decimal place was reported as
86.3584323 per cent. How should the result be reported?
6. The following results were obtained in the determination of chloride
ion in a sample of pure potassium chloride: 47.48, 47.56, 47.50, 47.62 and
48.25 per cent. Calculate: (a) the mean deviation, (b) the per cent deviation,
(c) the mean deviation in parts per thousand, (d) the mean error in per cent.
Should any of the results be rejected? Why?
16
QUANTITATIVE PHARMACEUTICAL CPEMISTRY
6. Find the common logarithms of the following numpers: 100, 65,8542,

0.221, 0.00018.
7. Find the numbers (antilogarithms) which correspohd to each 'of the
following logarithms: 3.5448, 0.8250, 0.1260, 2.3979.
8. Using logarithms, multiply: (a) 21500 X 0.000332, (b) 0.0648 X
0.0008426, (c) (42.16 - 1.85)(36.20 + 12.82)(0.5444 X 0.lJ12).
9. Using logarithms, divide: (a) 0.01648 by 0.9472, (b) 20.04 by 140.80,
(c) 1020 by 12.64.
GENERAL OPERATIONS
Crucibles.-A variety of crucibles are used in quantitative

analysis. Those most commonly employed are made of highgrade porcelain. They withstand high
temperatures and are suitftble for use
in the ignition of most '.p.rugs and
precipitates. They are not suitable
for fusions because the glaze is attacked
by the flux, especially if the flux is
basic. Other crucibles consist of fused
silica, alundum, nickel, and p~atinum.
The Gooch Filtration Crucible.-The
Gooch filtration crucible (Fig. 3) is
designed for the separation1of precipitates by suction filtration. It has a
perforated bottom upon which is
bedded a mat of asbestos, thus makFIG. a.-Diagram of Gooch ing it possible to collect, wash, dry,
crucible assembly for filtrad
. h
. .
.
tion.
an weIg a preCIpItate III the saple
crucible.
Preparation and Use of Gooch Crucible.-Obtain about 0.5
Gm. of asbestos fiber (washed with hydrochloric acid) and shake
it thoroughly with 100 cc. of distilled water in a suitable flask.
Allow the asbestos to settle and decant most of the water containing the fine fibers. Add about 100 cc. of water and agitate
the mixture ~gain. Fit the Gooch funnel containing the crucible
in the mouth of a filter flask connected to a suction pump. Pour
the suspended asbestos mixture into the crucible in small successive portions applying suction gently after the addition of each
portion until a smooth mat not over 1.5 mm. thick is obtained.
Upon holding the crucible to a strong light, the holes in the
17
REMARKS AND .GENERAL DIRECTIONS
bottom should be just perceptible and the mat of asbestos should

appear uniform. Cover ~he asbestos with a perforated porcelain
disk (filter plate) and add a very thin layer of asbestos. Wash
the filter mat with 200 to 300 cc. of water or until no trace of
asbestos fiber passes into the washing. Place the prepared
crucible on a watch glass and dry it for 1 hr. in an oven at 120 to
130. Allow the crucible to cool in a desiccator, then weigh.
Repeat until the weight of the prepared crucible is constant
within 0.0002 Gm. for two successive weighings.
To use the Gooch crucible, place it in the funnel and apply
moderate suction. Pour the liquid from the precipitate into
the crucible, wash the precipitate in the beaker by decantation,
transfer the precipitate to the crucible and
complete the washing. The rod used to
guide the liquid into the crucible should
never be allowed to come in contact with
the asbestos mat, since it may break the
thin film above the filter plate. The filtrate
should be examined for traces of asbestos FIG. 4.-Fritted-glass
crucibles, low form and
fiber, and if such are found, it should be high form.
repassed through the filter.
A properly prepared Gooch filter may be used for a number of
successive determinations where the precipitates are coarse
grained and crystalline.
Fritted-glass Crucibles.-Fritted-glass crucibles (Fig. 4) have
fused in sintered or fritted-glass bottoms and are supplied in two
porosities, "medium" and" fine." Crucibles of medium porosity
are suitable for the filtration of flocculent and moderately fine
precipitates. Crucibles of fine porosity are suitable for the
filtration of fine precipitates such as mercuric or bismuth sulfides.
These crucibles are useful in analyses where the residue is t9 be
dried to constant weight at temperatures below 150C. The
crucibles should .be heated in an oven and allowed to cool to
room temperature in a desiccator.
Filtration and Washing Precipitates.-The most usual and
efficient method of washing a precipitate is by the process of
decantation. The liquid above the precipitate is poured along
a guide rod onto a prepared filter, some of the liquid and mORt
of the precipitate being allowed to remain in th~ container.
18
A given amount of wash water is then poured on the precipitate

and stirred vigorously. The precipitate is allowed to settle,
and the supernatant liquid is decanted as before. This process
is repeated three or four times. The precipitate is then washed
onto the filter, using a "policeman" if necessary to dislodge
any particles which adhere to the container wall. The filter
containing the precipitate is washed by causing a fine stream of
liquid from the wash bottle to fall upon the filter, always washing
from the top edge down, until the filter is about three-fourths
full. The filter is allowed to drain until practically empty, then
the rinsing is repeated. After about five such rinsings, 1 or
2 cc. of the final filtrate should be tested for completeness of
washing.
It should be borne in mind that it is better to wash a precipitate
a comparatively large number of times with small portions of
liquid than only a few times with large portions and that each
portion of wash liquid should be removed as completely as
practicable before the next portion is added. Never fill the filter
more than to within one-fourth inch of its uppe];...edge. The size
of the filter should be governed by the amount of precipitate,
not by the amount of liquid to be filtered.
Colloidal and Fine-grained Precipitates.-Colloidal precipitates, some 'of them gelatinous, some of them finely divided,
are given by the most insoluble substances, such as sulfides,
liydroxides, elementary sulfur, etc. Although insoluble, they
have the property of being converted by water into colloidal
suspensions which pass through the filter. These substances
can usually be coagu1ated and rendered filterable by boiling
them after the addition of an electrolyte, such as NH 4 NO s,
NH 4 CI, or HCI. It is. in part for this reason that prepared wash
liquors, rather than pure water, are used for washing certain of
these precipitates.
All crystallization is preceded by supersaturation. If supersaturation is high, crystallization will be spontaneous and the
crystals formed will be so fine that they will pass through the
filter. If the precipitating agent is 'added slowly with constant
stirring, high local supersaturation will be avoided, and the
crystals formed will grow in size to such an extent that they will
be retained by the filter. By using different concentrations, it
19
is possible to get the same salt crystallized out in crystals varying

in size from full-faced crystals down to the colloidal form.
Evaporation of Liquids.-The evaporation of liquids is best
carried out in porcelain dishes constructed for that purpose which
expose a large surface of liquid to the air. Evaporations should,
with few exceptions, be carried out on the water bath to avoid
danger of loss of material due to spattering or bumping and to
avoid decomposition. The dish should be covered by an inverted
funnel supported above it or with a watch glass elevated above
the rim of the dish by means of a glass triangle to prevent the
entry of foreign matter into the liquid. Large quantities of
liquid are best evaporated in a comparatively small dish by
adding fresh portions of the original liquid from time to time
to replace that lost by evaporation. This helps to eliminate the source of error involved in the frequent transfer of
liquids.
Transfer of Liquids.-As few transfers of liquid as possible
should be made, since each permits of a possible source of error.
When transfers are necessary, they must be made quantitatively,
washing the original container with successive small portions of
wash liquid. This procedure is the most efficient, since it most
thoroughly removes the material and keeps the volume of liquid
within workable limits.
When transferring liquids from one' vessel to another, a guide
rod should always be used. It should be held tightly against
the lip of the container.
Drying and Ignition of Samples and Precipitates.-Many
substances are directed to be dried under standard conditions
before analysis to correct for absorbed moisture. ~recipitates
frequently must be dried previous to ignition. Numerous
types of drying ovens are marketed which are suitable for this
purpose, the electrically heated op.es being best suited to constant
temperature control. Samples should be dried at the specified
temperature on a watch glass or in a suitable container. Precipitates may be dried directly on a filter by placing the funnel
containing both directly in a drying oven adjusted to a temperature of 90 to 100 unless otherwise specified. The funnel
should be covered with a sheet of filter paper held in place by
crimping its edges over the rim of the funnel.
20
Precipitates may frequently be ignited without previous

drying. To do this, the precipitate is folded within the filter
paper, and the whole placed in a vertical crucible. The crucible
is covered and heated gently by intermittent application of the
flame until all of the moisture is driven off and the paper is
well charred. The cover is then removed, and the crucible
placed in an inclined position on the triangle (Fig. 5). The
full flame is then turned on and ignition continued until all carbon
FIG. 5.-Position of crucible above flame during ignition.
has been burned away, rotating the crucible slightly from time
to time to expose fresh portions of precipitate. The crucible
containing the ignite'd residue is allowed to cool in a desiccator
and weighed. It is then ignited a second time and weighed. If
the successive weighings do not agree within 0.2 mg. when a
porcelain crucible is used, it should be ignited and weighed a
third time. These operations should be repeated until constant
weight is attained.
Constant Weight.-The. term dried to constant weight means
that two consecutive weighings do not differ by more than 0.1
per cent when the second weighing is made after an additional
hour of drying.
21
Use of Desiccators.-Desiccators are a special form of

glass vessel, rendered air-tight by means of ground contact surfaces, used to maintain a dry atmosphere for objects that might
be affected by moisture or carbon dioxide. If the contact surfaccs are not air-tight, they may be made so by coating them
with a thin layer of grease prepared by melting together equal
parts Of beeswax and petrolatum. The desiccator is usually
divided into two compartments separated by a perforated porcelain plate. The lower compartment is charged with a dehydrating agent, one of the most common of which is anhydrous
calcium chloride, but concentrated sulfuric acid, solid potassium
hydroxide, unslaked lime, magnesium perchlorate, etc., are
sometimes used. The porcelain plate is usually fitted with
holes for crucibles. The desiccator should always be kept closed
when not transferring crucibles into or from it.
THE ANALYTICAL BALANCE
A knowledge of the principles and use of the analytical balance

(Fig. 6) is essential in all quantitative chemistry. The diagram
(Fig. 7) shows the various parts of such a b'alance with the proper
name of each part. Other balances on the market differ slightly
in construction but the parts are essentially the same for all.
The analytical balance is a delicate instrument of precision which
must be handled with extreme care to maintain its fine adjustment and accuracy. Each student under the direct supervision
of an instructor should learn the name and function of each part.
The following brief comments will serve to assist the student
better to understand the function and care of the balance: The
balance beam is supported on a center knife edge or fulcrum,
usually of agate, which rests upon an agate plate. Two pans
intended to support the masses to be compared are suspended
vertically from each end of the beam by stirrups. Each stirrup
has an agate bearing which res'ts on a knife edge. The arms of
the balance beam are graduated so that a rider, of known weight,
can be placed at any desired distance from the central knife
edge. Forcible or sudden lowering of the beam upon the knife
edges dulls them and produces furrows in the bearings which
decrease the sensitiveness of the balance. A pan arrest is
provided which when pushed in and turned releases the pans,
22
FIG. 6.-A typical analytical balance.

RIDER ROD CARRIER----_
CENTREKNIFEEDGE------_,
\
RIDER HOOK--__
STIRRUP-- __
------RIDER ROD
BEAMARREST--
STlRRUPHOOK---
GRAVITY WEIGHT
_____ COLUMN
____INPlCATOR
BowWIRES-
(OR. NEEDLZl
~ __ --.LEV)!:L
PAN---__
.---PAN ARREST
---INPEX PLATE
FIG. 7.-Diagram showing parts of the analytical balance.
23
leaving them suspended. from the stirrup. A long pointer which

multiplies the rotational displacement is attached to the beam in
order to render small movements of the beam perceptible.
Rules for the Use and Care of the Balance:
1. Use the balance assigned by the instructor.
2. Adopt an attitude of personal responsibility for the condition of your balance; the carelessness of one student may render
inaccurate the work of all who use the same balance.
3. Brush the pans and floor of the balance with a camel's-hair
brush before starting to weigh any substance.
4. Test the adjustment of the balance before each weighing.
The balance is properly adjusted only if the following conditions
are fulfilled: (a) The balance is level as shown by the spirit
level; (b) the mechanism for releasing and arresting the beam
works freely and smoothly; (c) the pan arrests just touch the
pans when the beam is lowered; (d) the pointer rests at zero
when the beam is either released or arrested while the pans are
supported; and (e) the pointer swings equal distances on either
side of zero on the index plate when the beam is set in motion
without a load on the pans or with an equal load on each pan.
Allowance may be made for a variation of one division on the
index scale without adjustment.
5. Do not attempt to adjust the balance. Call an instructor
if it requires adjustment.
6. No sample should ever be placed directly upon the balance
pan. Solids may be weighed on a watch glass or in a weighing
bottle, and liquids must always be weighed in a tightly stoppered
bottle. The use of sheets of paper in place of a proper container
is not permissible.
7. Hot objects must be allowed to cool to room temperature
before placing them inside the balance case, since currents of
warm air tend to buoy up one arm of the balance and also to
cause that arm to expand in length.
8. Material spilled on the balance pans or floor must be
brushed up at once.
9. The balance door must be closed when the final weighing is
made.
10. The beam should be set in motion by gently lowering it
so that the pans are supported by the pan arrests; the pan arrests
24
are then carefully lowered, and if the beam does not swing, it
may be set in motion by means of the rider. Long swings of the
pointer are not necessary. The pointer should swing through an
amplitude of not less than two divisions to either side of the zero
of the index scale.
11. All weights should be handled with the forceps; they should
never be touched with the fingers. Place heavy weights in the
center of the pan to prevent oscillation.
12. Record all weights directly into your notebook upon completion of a weighing. Before removing weights, check the
balance by arresting and releasing the beam and be sure that
correct balance has been attained. Remember that a slight error
in weighing may render worthless all further analysis of a sample.
13. Before leaving the balance, stop the motion of the beam
by means of the pan arrests, raise the beam off the agate knife.
edges, be sure the balance is clean, and close the door of the
balance case.
Sensitivity of the Analytical Balance.-The sensitivity varies
inversely with the weight of the beam. The lighter the beam,
therefore, the greater the sensitivity. The sensitivity varies
with the length of the beam. The sensitivity varies directly to
the time period of oscillation. The sensitivity varies inversely
with the load on the pans. The sensitivity of a balance may be
varied by moving the small gravity weight on the pointer upward
or downward, thus changing the position of the center of gravity
of the beam. The efficient'operation and sensitivity of a balance
are dependent upon minimum friction between the agate
knife edges and bearings.
Exercise 1
Object.-Determination of the True Zero Point of a Balance.

Procedure.-Dust the pans and floor of the balance with a camel's-hair
brush. Close the balance door. Gently release the beam and pan arrests,
avoiding setting the beam in motion. Cause the beam to swing so that the
pointer moves past not more than seven or less than two divisions of the
index scale by carefully touching one arm of the beam with the rider.
Allow the pointer to complete two oscillations without taking a reading.
Then take three readings of the limit of pointer oscillation on one side
of the index scale and two on the other, counting first left then right. Take
the average reading for each side, find the difference between the averages,
25
and divide t.he difference by two; the result gives the true zero point of the
balance.
The zero point is then com:euted as follows:

Right
Left
5.2
4.6
5.1
4.4
5.0
-3)15.3
2)9.0
5.1
4.5 average
5 . 1 - 4. 5 = o. 6 difference
0.6/2 = 0.3 of a scale division to the left which is the true zero
point. Make at least three such zero point determinations. The
results should check within two-tenths of a scale division.
The true zero point of a balance varies from day to day and
should be determined each time the balance is used.
Exercise 2
Object.-Determination of the Sensitivity of a Balance.

Procedure.-The sensitivity of a balance is the displacement of the zero
point produced by a weight of 1 mg. Determine the true zero, or rest,
point of the balance. Place the rider on the first division of the beam scale
and determine the new resting point on the index scale.
The difference between the two rest points gives the sensitivity
of the balance in terms of index scale divisions. Assume that the
difference in the rest points is two, then 1 mg. causes a displacement of two scale divisions. Since the index scale can be read
to one-tenth of a division and there are 20 one-tenth divisions,
the balance is said to be sensitive to 0.05 mg.; that is, the smallest
weight which can cause a readable deflection is 0.05 mg.
Determine the sensitivity of the balance with loads of 10 Gm.
on each pan and with 20 Gm. on each pan. Tabulate your
results.
WEIGHTS
Figure 8 illustrates a set of analytical weights. Analytical

weights may be purchased in graduated sets; a set in which the
26
QUANTITATIVE PHARMACEUTICAL CHEMISTRY~
largest weight is 50 Gm. and the smallest 5 mg. is satisfactory

for most analytical work. The 1 Gm. and all larger weights
are usually made of brass and plated with gold or coated with
lacquer to prevent corrosion. The fractional weights are usually
made of platinum, tantalum, or aluminum.
FIG. 8.-Set of analytical weights.
Comparatively accurate weights can be purchased, but an

analyst must know that his weights are accurate and should,
therefore, always calibrate the weights at least once each year.
Exercise 3
Object.-Calibration of a Set of Weights.

Procedure.-Clean the set of weights to be calibrated by wiping them
thoroughly with a clean cloth and brush the weight box with a stiff brush.
Mark duplicate weights by punching one or two dots on them so that
they may be readily distinguished. Determine the zero point of the balance. Place the 5 mg. weight on the left pan and the rider on the 5 mg.
position on the right arm of the beam scale and determine the zero point
again. Transfer the 5 mg. weight to the right-hand pan and place the 10
mg. weight on the left-hand pan and secure perfect balance by means of the
rider. Proceed in like manner with the remaining weights. Always place
the unknown weights on the left pan and the known weights on the right
pan. Tabulate the data according to the following illustration:
27
CALIBRATION CHART
Left
0 . 005
0.010
0 . 010
0.020
0 . 050
0 . 100
0.100
0 . 200
0.500
1.0
1.0
1.0
2.0
5.0
10.0
10 . 0
20 . 0
50.0
Right
Rider
0.005
0.01
0.01 + 0.01
0.02 + 0.01 + etc.
0.05 + 0.02 + etc.
O. 05
O. 02 + etc.
0 . 100 + 0 . 100
0 . 200 + etc.
0.500 + etc.
0.500 + etc.
0.500 + etc.
1 + 1
2+1+1+1
5+2+1+1+1
5+2+1+1+1
10 + 10
20 + 10 + 10 + etc.
Rider
reading
Actual
weight
Aliquot
Actual
part correction
+0 . 0050
+0 . 0052
+0 . 0001
-0 . 0002
+0.0042
+0 . 0045
+0 . 0044
+0 . 0008
+0.0044
+0.0040
+0 . 0024
+ 0.0034
-0 . 0014
0.0000
-0.0064
- 0.0052
-0.0024
-0 . 0032
0.0050
0.0102
0 . 0103
0.0203
0.0500
0.1003
0.1002
0.2013
0 . 5020
1.0036
1.0020
1.0030
2 . 0042
5.0128
10.0192
10.0204
20.0372
50.0992
0 . 0050
0.0100
0.0100
0.0200
0.0501
0.1002
0 . 1002
0.2004
0.5010
1.0019
1.0019
1.0019
2 . 0038
5.0096
10.0192
10.0192
20 . 0384
50.0950
0.0
+0.0002
+0.0003
+0.0003
+0.0001
+0.0001
0.0000
+0.0009
+0.0010
+0.0017
+0.0001
+0.0011
+0.0004
+0 .0032
0 . 0000
+0 . 0012
-0 . 0012
+0 . 0042
The information secured in the first four columns is all that

is required for a calibration of the weights in terms of relative
units. This method of calibration is sufficiently accurate for all
ordinary quantitative work . The calculation of the actual
correction is found by taking the 10 Gm. weight in terms of the
preliminary standard and finding the aliquot part of this weight
represented by each of the other weights. In our table the actual
weight of the 10 Gm. weight in terms of the preliminary standard
was found to be 10.0192; the aliquot part of the 5 Gm. weight
will then be 10.0192/ 2 = 5.0096; the aliquot part of the 2 Gm.
weight will be 10.0192/5 = 2.0038; and the aliquot part of the
1 Gm. weight 10.0192/10 = 1.0019; etc. The aliquot part of
10.0192 is represented in the fifth column as calculated above.
In the sixth column are given the actual corrections as plus or
minus depending on whether the actual weight is greater or less
than the aliquot part.
The cOTrections in tenths of a milligram should be tabulated
on a card, and the card fitted tightly into the cover of the weight
box.
28

Weight,
grams
0.005
0.010
Correction,
0.1 mg.
0
+2
O.OlO
+3
0.020
0.050
0.100
0.100
0.200
0.500
+ 3
0
+ 1
0
+9
+10
Weight
1.0
1.0
1.0
2.0
5.0
10.0
10.0
20.0
50.0
Correction,
0.1 mg.
+17
+ 1
+11
+ 4
+32
0
+12
-12
+42
Riders.-Riders are supplied in 5 and 10 mg. weights. Always

check the weight of a new rider before weighing against the
0.005 Om. weight.
1. Why must the qualitative tests precede the assay to obtain accurate
data relative to the purity of an official substance?
2. What is a Gooch crucible? Why should asbestos fiber washed with
hydrochlori,c acid be used in preparing this crucible for use in filtration?
3. What is a desiccator? What two agents are used most fommonly to
dehydrate official samples before assay?
4. What factors affect the sensitivity of a balance?
6. Why are agate knife edges and agate supports used in balances?
6. Why should the true zero point of a balance be determined before
each weighing?
'1. Explain how the sensitivity of a balance may be determined with a
load of 10 Gm. on eMh pan.
CHAPTER II
GRAVIMETRIC ANALYSIS
THEORY
The student should be familiar with the modern chemical

theories which apply in quantitative analysis from the study of
general chemistry and qualitative analysis. In quantitative
analysis, theoretical considerations must be understood and
applied in order to know the full explanation of what takes place
in an analysis. A brief review of the fundamentals of the various
theories with their quantitative application will be given in
conjunction with those procedures to which they apply.
The reactions of quantitative analysis take place in accordance
with the established laws and theories of chemistry, i.e., theory
of ionization, law of mass action, common ion effect, reversible
reactions, and solubility product principle.
Theory of Ionization.-The theory of ionization is based on
the following postulates:
1. Electrolytes in solution form ions. When electrolytes are
dissolved in water, their molecules dissociate or break up into
particles termed ions; thus, when hydrogen chloride gas is dissolved in water, some of the hydrogen chloride molecules dissociate or ionize to form hydrogen ions and chloride ions so that
from one molecule of hydrogen chloride, two ions are obtained.
2. Some of the ions are charged positively, others negatively;
the sum of positive and negative charges always being equal, the
solution remains electrically neutral. In the case of hydrogen
chloride, the hydrogen ions are charged positively and the
chloride ions negatively, the sum of positively charged hydrogen
ions being equal to the sum of negatively charged chloride ions;
the solution of hydrogen chloride gas remains electrically neutral.
3. The properties of the ions into which the electrolyte dissociates differ from the original substance. The hydrogen and
chloride ions have properties different from the original hydrogen
29
30
chloride gas, and the hydrogen and chloride ions differ from
hydrogen and chlorine gas, respectively.
4. The ionization of the electrolyte results in an equilibrium.
The point at which equilibrium is reached is dependent on the
nature of the electrolyte, the nature of the solvent, and upon
the dilution; thus, electrolytes differ in the extent to which they
ionize; some solvents, such as ether and benzene, produce no
ionization, but in solvents which produce ionization, the greater
the dilution the greater the extent of ionization of the electrolyte.
A certain state of equilibrium exists between the ions and
undissociated molecules for every degree of dilution. The
equilibrium'''may be represented as follows: NaCI
Na+ + Clwhen sodium chloride is dissolved in water. Since the extent
of dissociation is dependent on dilution, the greater the dilution
of the solution the more sodium chloride will dissociate into ions
until at infinite dilution it may be regarded as completely ionized.
'l'b . .
db h
. [Na+J X [Cl-J
The eqUlI
num IS represente y t e equatIon
[NaCl]
=
K where [Na+], [Cl-], and [NaCl] represent the concentration

of sodium ion, chloride ion, and sodium chloride, respectively,
and K represents the ionization constant.
Reversible Reactions.-Most of the reactions iAvolved in
quantitative analysis are of the reversible type. Under certain
conditions they may be made to continue to completion, but
under other conditions they may attain equilibrium before
completion, resulting in loss of a portion of the substance being
analyzed. It, therefore, is very important to understand what
conditions must be satisfied to make the reaction go forward
to completion so that it will be of value in quantitative work.
Thete are three general conditions which tend to destroy equilibrium and lead to complete reaction: (1) the formation of an
insoluble gas; (2) the formation of a sparingly soluble solid;
(3) the formation of very slightly ionized molecules.
According to the law of mass action, the speed of a reaction is
proportional to the products of the molecular concentrations of
the teacting substances. Since the speed of a reaction depends
upon the concentration of every substance taking part in the
reaction, the point of equilibrium will depend upon the concentration of each of the components of the two opposing reactionsj
31
e.g., in the reaction KNO a H 2S0 4

KHS0 4
HNO a, the
speed of the reaction of potassium nitrate with sulfuric acid is
_
expressed by the equation:
Speed = (KNO a] X (H 2S0 4] X k where k is the affinity constant of the reaction.
The speed of the opposing reaction is expressed by the equation:
Speed = [KHS0 4 ] X [HNOa] X kl where kl is the affinity constant of the opposing reaction. At equilibrium the speeds of
the two reactions are equal.
and
Since kl and k are constants, their quotient K is a constant

termed the" equilibrium constant."
At a definite temperature the equilibrium constant is a fixed
value for any given reaction irrespective of the concentration
of the substances present. If, therefore, the concentration of
sulfuric acid is increased, all of the other concentrations must
change, the concentration of KNO a must become less, and that
of both KHS0 4 and HNO a greater to maintain the equilibrium
constant, with the result that equilibrium is forced to shift
toward the right. In quantitative analysis, an excess of one
component is frequently added to cause the reaction to go as
nearly to completion as possible.
Solubility Product Principle.-The solubility product principle is an application of the law of mass action to equilibria
which generalizes the behavior of difficultly soluble salts in their
saturated solutions. Whenever precipitation occurs, is prevented, or solution effected, the conditions to which this principle applies are involved. Tlte principle may be stated as
follows: The product of the concentration of the constituent ions in a
saturated solution of a difficultly soluble salt for any given temperature is practically a constant, each concentration being raised to a
power equal to the relative number of ions supplied by one molecule
of the salt upon dissociating, i.e., a difficultly soluble salt AmBn
32
upon dissociating would furnish a relative number of m cations

and n anions:
AmBn
mA+ + nB- and, using C to designate concentration, the solubility product would be (C A +)'" X (CB_)n = solubility product AmBn.
The solubility products of some of the more difficultly soluble
salts dealt with in pharmaceutical analysis are given in the
following table:
TABLE I.-SOLUBILITY PRODUCTS OF SOME IMPORTANT SALTS
Substance
Aluminum hydroxide ....... .

Barium carbonate . ....... .
Barium sulfate . ........... .
Calcium carbonate ........ .
Calcium oxalate ........ "
Lead carbonate .......... .
Lead sulfate. . . . . . . .. . .... .
Magnesium ammonium phosphate ................... .
Magnesium hydroxide ... .. .
Magnesium oxalate ........ .
Mercuric sulfide .......... .
Mercurous chloride ........ .
Silver bromide ............. .
Silver chloride ............. .
Silver iodide ............... .
Silver thiocyanate ......... .
Temperature,
C.
25
25
25
25
25
18
18
Ions involved
AIO,- X H+
BaH X CO,-Ba'+ X SO.-Ca++ X CO,-Ca++ X C,04-Pb H X CO,-Pb++ X S04--
18
18
MgH X NHL': X PO.--Mg++ X (OH-),

MgH X C,O.--
25
25
25
25
25
25
Hg++
Hg+
Ag+
Ag+
Ag+
Ag+
25
X
X
X
X
X
X
S--
crBrCl1SCN-
Solubility
product
3.7
8.1
1.08
9.3
2.6
3.3
1
X
X
X
X
X
10-16
10'"
10- 1
10'"
10'"
2.5
X 10-13
3.4
2.6
4
3.5
7.7
1. 5
0.9
1. 2
X
X
X
X
X
X
X
X 10-14
X 10'"
10- 11
10'"
10-54
10- 18
10-1 '
1{)-10
10- 18
X 1{)-12
When a number of different kinds of ions are present in the

same solution, the greatest concentration that anyone of them
can attain is determined by the others. Thus, no great concentration of Ag+ can be present in a solution in the presence of
CI-, for the two ions unite to form a precipitate of the difficultly
soluble salt, silver chloride. Silver chloride is slightly soluble
in water, the solubility being about 0.00001 mole (1.5 mg.) per
liter. When this solubility is exceeded, a precipitate of Agel
forms which is in equilibrium with the dissolved silver chloride.
When the equilibrium is established, the supernatant liquid is a
saturated solution, and the tendency of the solid to go into
solution is exactly equal to the tendency of the dissolved salt
33
to precipitate. When this condition exists, the following scheme

of equilibria is set up:
AgCl
AgOl
Ag+ + 01solid dissolved dissolved
ppt. unionized
ionized
The ionization equilibrium expressed according to theory is
[Ag+] X [01-] ionized
'"
[A g 01] UlllOlllze
. . d
= K = IOnIZatIOn constant
Since the solution is saturated with AgOI at a given temperature and the concentration of unionized [AgOl] remains constant,
it follows that the product K X [AgOI] also remains constant
and that in a saturated solution of a difficultly soluble salt, the
product of the concentrations of its ions is constant.
In the calculation of the solubility product, it is customary to
express the concentration in terms of moles per liter. A saturated
solution of silver chloride contains 1.5 X 10-3 Gm. (0.0015 Gm.)
per liter. The molecular weight of silver chloride IS 143.34.
In terms of molarity, therefore, the solution contains
1. 5 X 10143.34
1 1 X 10-5
moles per liter. At this dilution the dissolved silver chloride may
be assumed to be completely ionized, AgOI~Ag+
01- so
that each mole of silver chloride furnishes 1 mole of silver ions
and 1 mole of chloride ions. The solubility product, S.P., then
[Ag+]' X [01-] = (1.1 X 10- 5) X (1.1 X 10- 5)
= 1.2
X 10-1 ,
and the ionic product is equal to the solubility product.

If the solubility of a compound is known, the solubility product
may be calculated; e.g., a saturated solution of silver iodide at
250. contains about 0.00235 mg. of AgI per liter. The molar
solubility is equal to the solubility in grams divided by the
gram-molecular weight, or
~
0.0~~~~8235 =
0.00000001
= 10-8
Assuming 100 per cent ionization at this dilution, the concentrations of Ag+ and of 1- also equal 10-8 , since each molecule of
34
AgI forms 1 Ag+ ion and 1 I- ion, and the solubility product is
equal to the ionic product
Ou+ X Ox- = 10-8 X 10-8 = 10-16 = S.P. of AgI
From the above calculation of solubility product, it can be
predicted that silver iodide will precipitate if Cu+ X 0 1- becomes
greater than 10- 16 ; that in a supersaturated solution Cu + X Oxbecomes greater than 10- 16 ; that in a saturated solution OAa+
X Ox- wilJ..be equal to 10-16 ; an(that to dissolve precipitated
AgI, OAa+ X 0 1- must be less than)O-16.
In the above illustration, the solubility product of silver iodide
was calculated from solubility data. Conversely, if the solubility
produce is known, the solubility of a compound may be calculated. In the table, the solubility product of silver chloride is
given as 1.5 X 10- 10 at 25C. How much silver chloride will
dissolve in 100 cc. of water at the above temperature?
S,P' AgCI = GAg+ X GCI- = 1.5 X 10- 10
If OAg+ = x,
then
OCl- = x,
since
OAg+
x 2 = 1.5 X 10-10
X = 1.22 X 10-5
= OCl-
x is expressed in terms of moles per liter. Since lthe solubility

of AgOl is equal to 1.22 X 10-5 moles per liter, the solubility
1.22 X 10-5
in grams per 100 cc. is equal to
10
tImes the gram6

molecular weight of AgOl or 1.22 X 10- X 143.5 = 0.000175
Gm. AgCl soluble in 100 cc. of water.
The solubility product is thus seen to be an ultimate value
attained by the ionic product when equilibrium has been established between the undissolved solid and the difficultly soluble
salt in solution. If the product of the concentration of any
pair of ions in solution is made to exceed in value the solubility
product of the compound formed by their union, precipitation of
the compound will take place until the product of the ionic
concentration is exactly equal to the solubility product value,
and when the product of the ionic concentrations is made less
than the solubility product value, the compound formed by
their union will dissolve until the product of ionic concentrations
is equal to the solubility product value.
35
Common Ion Effect.-The equilibrium constant does not

change no matter what the concentration of the reacting substances may be. The relative concentration of the reacting
substances may change but there is no change in the equilibrium
constant. When two substances furnish an ion in common so
that the concentrations of positive and negative ions of an electrolyte are unequal, the law of mass action causes equilibrium
to be maintained; thus, when a solution of silver nitrate is added
to a solution of sodium chloride, the chloride ion is momentarily
present in a concentration such that its ionic product with the
silver ion exceeds the solubility product of silver chloride, and the
insoluble silver chloride is precipitated:
Ag+
+ Ol---l-AgOl ~
When an equivalent amount of silver nitrate has been added,

and the system has acquired equilibrium, the concentration of
silver ions will be exactly equal to the concentration of chloride
ions.
If to the supernatant liquid which is a saturated solution of
silver chloride a small amount of a soluble silver salt or a soluble
chloride be added, a slight further precipitation will take place.
It follows from the application of the equilibrium representing the
ionization constant
[Ag+] X [01':"] = K
[AgOI]
that if the concentration of silver ion be increased by the addition
of a soluble silver salt, the concentration of chloride ion must
decrease and, conversely, that if the concentration of chloride
ion: be increased by adding a soluble chloride, the concentration
of silver ion must decrease since their product remains constant.
This decrease in the concentration of the ions in either case can
be accomplished only by the ulJion of silver and chloride ions to
form insoluble silver chloride forcing the reaction toward
completion.
The common ion effect is used frequently in gravimetric
pharmaceutical analysis to drive reactions toward completion.
Other examples of COmIllon ion effect will be discussed in conjunction with the assays in which they occur.
36

1. What are the postulates upon which the theory of ionization is based?
2. What is a reversible reaction?
3. What conditions tend to destroy equilibrium and cause a reaction to
go to completion?
4. Define and illustrate the solubility product priilCiple.
5. 0.247 mg. of barium sulfate dissolve in 100 cc. of water at 25C.
Express the solubility in terms of molar solubility.
6. From the solubility of BaSO. given in'problem 5 calculate its so_lubility
product assuming 100 pel' cent ionization.
7. The solubility product of calcium sulfate is approximately 2 X 10- 4
Calculate the amount of calcium sulfate soluble in 500 cc. of water. Assume
100 per cent ionization.
S. Illustrate how common ion effect may be utilized to secure quantitative
precipi tation.
9. A saturated solution of lead chloride at 20C. contains 0.07 mole per
liter. This solution is 80 per cent ionized. Calculate the solubility product.
CHAPTER III
GRAVIMETRIC METHODS
Gravimetric analysis implies that the substance to be determined is to be separated from a weighed sample in the form of a
compound of known composition and weighed. Knowing the
weight of the original sample and that of the product, the weight
and percentage of any component common to both can be calculated. The product to be weighed in pharmaceutical analysis
may be obtained by any'one of various methods: (1) It may be
precipitated from solution; (2) it may be the decomposition
product resulting from ignition of a compound; (3) it may be
deposited on an electrode by electrolysis; (4) it may be separated
from other substances by extraction with a solvent; and (5) it
may be obtained by absorbing a gas in some substance of known
weight and finding the increase in weight produced by the absorption of the gas. The first two methods comprise the subject
matter considered in this chapter.
Exercise 4
Determination of Chloride Ion in a Soluble Chloride.-Chloride
ion is determined gravimetrically by precipitating and weighing it
as silver chloride. An excess of solution of silver nitrate, slightly
acidified with nitric acid, is added to the solution of soluble chloride. The precipitate is filtered out, washed, dried, and weighed
as silver chloride. Other substances which yield insoluble silver
salts must be absent.
No method for the gravimetric determination of chloride ion is
official, but the procedure constit.utes a classic example of gravimetric analytical technique with which every student should be
familiar.
Object.-Assay of Sodium Chloride.
Materials Required.-O.25 Gm. of sodium chloride.
5 per cent silver nitrate solution.
37
38
Diluted nitric acid.

Asbestos fiber.
Procedure.-l. Accurately weigh two samples of 0.2 to 0.3 Gm. of sodium
chloride.
This is most easily done by weighing the sample tube (weighing

-bottle) and contents accurately to 0.1 mg., without touching it,
entering the weight in the notebook. Hold the weighing bottle
directly above a 300 to 400 cc. beaker, labeled I, remove the
stopper, and carefully transfer 0.2 to 0.3 Gm. of the sample to
the beaker. Replace the stopper and again weigh the weighing
bottle to 0.1 mg., entering the weight in the notebook. The
first weight less the second is the weight of sample taken. In
like manner weigh a second 0.2 to 0.3 Gm. sample into beaker II,
entering the weights as before. Treat each sample as follows:
2. Completely dissolve the sample in 100 to 125 ee. of distilled water.
Add about 1 cc. of diluted nitric acid slowly with stirring. Test the solution
by touching a strip of blue litmus paper with the moist stirring rod. If it
is not acid in reaction, add sufficient acid to make it so. Measure out 5 cc.
in excess of the amount of silver nitrate solution theoretically required to
precipitate all of the chlorine as silver chloride. 4.dd the silver nit.rat~
solution in small successive portions, stirring constantly with a suitable
glass rod. Cover the beaker with a watch glass, and, '\fith occasional
stirring, gradually heat the mixtme to the boiling point. Turn down the
flame and digest the mixture, without boiling for 10 min. This treatment
coagulates the precipitate, causes it to settle, and leaves the supernatant
liquid clear. Add 1 or 2 drops of silver nitrate solution to,'the hot supernatant liquid to test for complete precipitation. If a precipitate forms, add
5 cc. more silver nitrate solution, stir, allow to settle, and test as before.
Set the beakers aside away from direct sunlight to allow the precipitate to
settle.
The solution is acidified with nitric acid to prevent the precipitation of other substances insoluble in water but soluble in nitric
acid such as carbonates, oxides, and phosphates. The acid also
helps to coagulate any colloidal silver chloride. Too great an
excess of nitric acid must not be added, since it exerts some solvent action on silver halides. The acidified solution should not
be heated until the silver nitrate has been added, since chlorine
may be liberated and lost, thus:
NaCl
6HCl
+ HNO a---7HCl + NaNOa
+ 2HN03~3C12 + 4H 20 + 2NO
GRA VIMETRIC METHODS
39
The silver nitrate is added in excess to drive the reaction to

completion through the production of common ion effect, assuring
complete precipitation:
AgNO s
+ NaCl---+AgCl t + NaNOs
The precipitation must be performed away from strong light,

since silver chloride is decomposed by sunlight with loss of
chlorine, while in diffused light the error from this source is
negligible.
The precipitate is digested near the boiling point to coagulate
colloidal silver chloride which, if present, would pass through the
filter.
3. During intervals in the above procedure, carefully clean and prepare
two Gooch crucibles for filtration as previously described. After the crucibles have been properly prepared, heated to constant weight, and weighed,
fit them into the suction flask. Be sure that the crucibles can be identified
as I and II by a suitable mark. Decant most of the supernatant liquid
into the Gooch crucible, using gentle suction to hasten filtration, guiding the
liquid into the crucible by means of a glass rod held against the lip of the
beaker. Wash the precipitate three times with 15 cc. portions of hot wash
solution prepared by adding a drop or two of dilute nitric acid to 200 cc. of
distilled water.
In pouring the clear liquid through the filter, retain as much of

the precipitate as possible in the beaker. When the third wash
solution has been added to the precipitate, transfer the precipitate
into the Gooch crucible using a stream of hot wash solution from
the wash bottle and a "policeman 11 to dislodge all particles which
adhere to the sides of the beaker and guide rod. Wash the
precipitate on the filter with several small portions of acidulated
wash solution, allowing each portion to run through before adding
another. Continue washing the precipitate in this manner until
1 to 2 cc. of filtrate give no test for silver upon the addition of a
few drops of dilute hydrochloric acid solution. When tested in
this way, the filtrate should gi.ve no greater turbidity than a
portion of the wash solution being used tested in the same manner.
4. When the filtrate is shown by the test to be free of silver nitrate, wash
the precipitate twice with 5 cc. portions of distilled water to remove most
of the nitric acid retained by the precipitate from the former wash solution.
Apply strong suction to remove as much liquid from the precipitate as
possible. Dry the crucible to constant weight at 110 to 120 0 in an air bath
40
or electric oven: until successive weighings differ by no more than 0.0002

Gm. Allow the crucibles to cool in a desiccator before weighing. The
weight of the crucible and precipitate mip.us the weight of the crucible gives
the weight of silver chloride obtained from ~he sample.
The precipitate is washed to remove soluble salts present in the

solution, chiefly sodium nitrate and excess silver nitrate. Hot
wash solution increases the solubility of these salts and keeps
the silver chloride from passing into the colloidal condition. The
solubility of silver chloride increases slightly in hot solution, but
this is compensated for by the smaller amount of wash solution
required.
Silver chloride is appreciably volatile when ignited, so it
should be dried at a relatively low temperature.
The reaction involved in the calculation of the amount of silver
nitrate solution to be used and of the purity of the sample IS
AgNO a + NaGI~AgGI + NaNO a
169.89
58.45
143.34
85.01
SAMPLE DATA FOR GRAVIMETRIC DETERMINATIONS
Weight of
II
Bottle before removal of sample ...

Bottle after removal of sample ....
Sample .........................
Crucible ........................
Crucible and silver chloride 1st ....
Crucible and silver chloride 2d ....
Crucible and silver chloride 3d ....
Crucible ........................
Silver chloride ...................
14.3580
14.0420
0.3160
18.2468
19.0204
19.0201
19.0200
18.2468
0.7732
14.0420
13.7315
0.3105
17.8827
18.6409
18.6403
18.6402
17.8827
0.7575
The equation' shows that 1 molecule of sodium chloride reacts

with 1 molecule of silver nitrate. From the molecular weights
NaGl = 58.45 and AgNO a = 169.89, therefore, the theoretical
amount of silver nitrate solution required to precipitate all of
the chloride ion as silver chloride can be calculated as follows:
169.89/58.45 = 2.9066 Gm. AgNO a required to precipitate the
chlorine from 1 Gm. of NaCl. Since, from the data, 0.3160
Gm. of NaCI were used, then 0.3160 X 2.9066 = 0.9185 Gm.
AgNO a required. Since the reagent silver nitrate is a 5 per
41
cent solution, each cubic centimeter must contain 0.05 Gm. of

AgNO s and the amount of solution required is 0.9185/0.05 =
18.37 cc. of 5 per cent solution.
The weight of the sample and the weight of silver chloride
obtained from it being known, the per cent purity of the sample
or the weight of chlorine in the sample may readily be calculated:
59.45/143.34 = 0.4078 Gm. NaCl equivalent to 1 Gm. of
AgCl. Since 1 Gm. of AgCl is equivalent to 0.4078 Gm. of N aCl,
then 0,7732 Gm. of AgCl found is equivalent to 0.4078 X 0.7732
= 0.3153 Gm. NaCl. The per cent purity of the sample is
0.3153
therefore 0.3160 X 100 = 99.78 per cent.
Having calculated the equivalent weight, the calculation of
per cent purity may be obtained by a single formula applicable
in all gravimetric determinations, thus:
WXE
--S- X 100 = per cent where W is weight of AgCl, E is
the equivalent weight, and S the weight of sample; e.g.,

0.7732 X 0.4078 X 100 = 9978
t
0.3160
. psr cen .
To calculate the per cent chlorine in the sample, substitute
the weight of chlorine, 35.45, for that of N aCl.
35.45/143.34 = 0.2474 Gm. Cl equivalent to 1 Gm. of AgCl.
Substituting this value for E in the above formula gives the per
cent chlorine in the sample.
The quantity by weight of an element or compound which is
equivalent to one part by weight of some other substance is
termed a chemical factor. For example, the ratio or factor
NaCl/AgCl = 58.45/143.34 = 0.4078 means that 0.4078 Gm. of
pure N aCl corresponds to 1 Gm. of AgCl. To find the amount
of N aCl represented by any given weight of AgCl, therefore,
the weight of AgCl obtained is multiplied by this factor. Thus
0.1 Gm. of AgCl is equiyalent to 0.1 X 0.4078 = 0.0408 Gm.
NaCl. Likewise,
Ag/AgCl = 107.88/143.34 = 0.7526. This
means that 1 Gm. of AgCl contains 0.7526 Gm. of silver and that
if we wish to calculate the weight of silver in a specific weight of
silver chloride, the latter need only be multiplied by this factor;
e.g., 5.2 Gm. of AgCl contain 5.2 X 0.7526 = 3.9136 Gm. Ag.
42
The gravimetric method for the determination of chloride ion

may be applied to estimate the purity of most soluble chlorides,
bromides, iodides, cyanides, thiocyanates, etc., as well as to the
standardization of hydrochloric acid solutions and the estimation
of silver compounds. Chlorates, perchlorates, and other oxyacids of the halogens may be determined by this method after
reducing them to the halides. The volumetric precipitation
method has superseded the gravimetric method because it can
be carried out more rapidly. The greatest objection to this
method is that impurities in the form of halogen salts are included
in the determination, the results being in reality a measure of the
halogens present.
1. Why should the solution be acidulated with nitric acid?
2. Why add silver nitrate in excess in the precipitation of the chloride?
3. Why is the precipitate washed with water slightly acidulated with
nitric acid?
4. Why is it best to wash the precipitate by decantation before it is
transferred to the Gooch crucible?
5. Why should th'is determination be carried out away from direct sunlight?
6. Name several official substances other than sodium chloride which may
be assayed by an analogous method.
7. What substances, if present, would interfere with the determination
of the purity of a soluble chloride?
8. In the precipitation of chloride from 0.600 Gm. of an unknown substance, a precipitate of 0.6800 Gm. of AgCI was obtained. Calculate the
per cent chlorine in the unknown.
9. How many grams of AgNO. are necessary to precipitate the chlorine
from 1 Gm. of 50 per cent zinc chloride solution?
10. When can it be assumed that a crucible is of constant weight?
11. Why are samples usually dried before portions are weighed for
analysis?
12. Give a rational procedure for the determination of silver by the
gravimetric method.
13. What is meant by the terms chemical factor and chemical equivalent?
Illustrate.
Exercise 5
Determination of Sulfate Ion in a Soluble Sulfate.-The
sulfate ion in a soluble sulfate may be determined gravimetrically
43
by precipitation as barium sulfate, the precipitate being collected,

dried, ignited, and wflighed.
Object.-Assay of Sodium Sulfate.
Materials Required.-l Gm. of sodium sulfate.
Barium chloride T.S. (12 per cent).
Procedure.-Weigh accurately about 0.4 Gm. of the dried salt, obtained
by drying about 1 Gm. to constant weight at 1200., dissolve it in 200 cc.
of distilled water, and add 1 cc. of hydrochloric acid. Heat to boiling, and
gradually add an excess of hot barium chloride T.S. Heat the mixture for
30 min. on a water bath, collect the precipitate of barium sulfate on a filter,
wash it until free from chloride, dry, ignite, and weigh. The weight of the
barium sulfate thus obtained, multiplied by 0.6086, indicates its equivalent
of Na.S04.
.
The solution of the sulfate is acidulated with hydrochloric

acid to prevent the precipitation of carbonates, etc., which are
soluble in hydrochloric acid solution. The acid also increases
the solubility of barium sulfate slightly and therefore promotes
the growth of large crystals, since small crystals dissolve more
rapidly than large ones. The small crystals expose more surface
area per unit of weight than do large ones. Consequently, they
dissolve more rapidly, and in a saturated solution the larger
crystals with small surface area exposed grow more rapidly than
they dissolve. The solution is heated to boiling to expel dissolved CO 2 An excess of hot barium chloride solution is added
to insure complete precipitation and to decrease the solubility
of barium sulfate by common ion effect. The mixture is digested
on a water bath for 30 min. to allow the larger crystals to grow
at the expense of the smaller ones, since the latter are more
rapidly soluble.
The barium sulfate is directed to be washed on a filter to
remove excess BaCh and the NaCl formed, but 'the washing
process may be carried out more rapidly by washing once by
decantation. The precipitate is then transferred quantitatively
onto an ashless filter by the same technique used in Exercise 4
and washed free of chlorides vas shown by tests made with silver
nitrate solution on 2 cc. portions of the filtrate. Always acidulate the filtrate with a drop of nitric acid in carrying out the test
for chlorides. Sometimes fine-grained crystalline precipitates
will pass through the filter. This may be observed by imparting
a rotatory motion to the filtrate, causing any crystals which
44
passed through to gather in the center of the beaker. If such

are detected, refilter.
Barium sulfate exhibits the property of "dragging down"
other soluble salts when it is precipitated. This property is
known as co-precipitation. The occluded or co-precipitated
salt cannot be removed by ordinary washing. The error from
this source is reduced largely by precipitation from hot dilute
solutions and by adding the precipitating reagent slowly with
continuous stirring. The phenomenon of co-precipitation is
particularly liable to occur when barium sulfate is precipitated
TABLE Ir.-OFFICIAL SUIlSTANCES ASSAYED BY GRAVIMETRIC PRECIPITATION
AS SULFATE
Substance
Weight
of
Ignition
sample, residue
Factor Official requirement, per cent
Gm.
--U.S.P.
Lead 8ubaceta teo R ..........
Sodium sulfate ..............
Sulfur ointment ..... ........
Sulfur (all forms) ...........

N.F.
Ichthammol for (NH.),SO ...
Ichthammol for total sulfur ...
Potassium guaiacol sulfonate.
Potassium sulfate ........ , ..
Sodium sulfate exsiccated ... .
R.
1.0
0.4
0.5
1.0
PbSO. .......
BaSO. 0.6086
BaSO. 0.1373
BaSO. 0.1373
1.0
1.5
1.0
0.6
0.5
BaSO.
BaSO.
BaSO.
BnSO.
BaSO.
0.5661
0.1373
1.03757
0.7465
0.6086
Pb = 70 to 73
Na.SO. = 99
S = 13.5 to 16.5
S = 99.5
(NH.),SO. = not more than 8
S = 10
C.H 3.OH.OCH 3SO.K = 97
K,SO. = 99
Na,SO. - 99
reagent.
in the presence of chI orates, nitrates, and the salts of iron,

aluminum, chromium, and other trivalent elements.
The precipita~e may be dried and ignited by the wet method as
described under General Operations (page 20). The equation
Na 2S04
142.05
+ BaCb.2H20~BaS04 + 2NaQI + 2H20

233.42
expresses the stoichiometric relationship on which the calculations are based:
;:~:~~
0.6086 Gm. Na2S04 equivalent to 1 Gm. of BaSO,
45
The gravimetric determination of sulfate ion as barium sulfate

is used extensively in pharmaceutical analysis. It may be
. applied to the estimation of metals which form insoluble sulfates as barium and lead. Sulfides and sulfur, free, and in
organic combination, may be oxidized to sulfate and subsequently precipitated as barium sulfate. Other compounds as
thiosulfates, sulfites, persulfates, alkaloidal sulfates, etc., may
be estimated by the application of similar procedures.
1. Why is the solution of soluble sulfate acidulated with hydrochloric
acid?
2. Why is it necessary to digest the precipitate of barium sulfate previous
to filtration?
3. Why should the precipitation of sulfates be carried out in hot, dilute
solution?
4. An unknown sample of a soluble sulfate weighing 1.8000 Gm. yielded
0.9000 Gm. of BaSO.. Calculate the percentage of sulfur in the unknown.
6. Calculate the equivalent amounts of each and the percentage purity
if the unknown in problem 4 were MgSO., K 2 SO., AI 2 (SO.)a, or H 2S0 4
6. Write the equations representing the reactions that take place in the
assay of sulfur ointment.
7. How much BaSO. would be formed from 1.2000 Gm. of Na 2SO . 10H 20?
8. What is the per cent of SO. in a sample of K.SO. if 0.5000 Gm. yields
upon reaction with BaCl 2 a precipitate weighing 0.5850 Gm.?
9. How is lead determined as sulfate? See reagent lead subacetate,
U.S.P.
Exercise 6
Determination of the Mercury Content of a Mercuric Salt.The mercury is precipitated as sulfide, washed, dried, and
weighed.
Object.-Assay of Mercuric Chloride.
Materials Required.-O.5 Gm. of mercuric chloride.
1 cc. of hydrochloric acid.
About 70 cc. of alcohol.
About 40 cc. of carbon tetrachloride.
Procedure.-l. "Dry about 0.5 Gm. of Mercury Bichloride to constant
weight over sulfuric acid, weigh accurately, and dissolve in 300 cc. of warm
distilled water to which 1 cc. of hydrochloric acid has been added. Pass
hydrogen sulfide through the cold solution until the precipitate of ~ercuric
sulfide readily subsides, leaving a clear, supernatant liquid."
46
The hydrogen sulfide gas used in the precipitation should be

washed by bubbling it through a gas-washing bottle containing
water. The glass delivery tube which passes to the bottom of
the precipitation beaker should be attached to the gas-washing
bottle by means of a rubber t.ube in such a manner that it can
be removed readily and held over the precipitation beaker to
wash any adhering precipitate into the beaker.
The hydrogen sulfide reacts with the mercuric chloride to form
a double salt of mercuric sulfide and mercuric chloride at first and
then this double salt is decomposed with the formation of black
mercuric sulfide as more hydrogen sulfide is passed into the mixture. Since mercuric sulfide is insoluble in acids, hydrogen
sulfide may be employed as the precipitating agent and hydrochloric acid may be added to repress the concentration of sulfide
ion so that metals which form sulfides more soluble than mercuric
sulfide and which may be present as impurities will not be
precipitated.
2. "Collect the precipitate on counterpoised filters or in a tared Gooch
crucible, wash it well with cold distilled water, and finally~with three portions
of about 10 cc. each of alcohol. Then close the tip of the funnel or of the
Gooch crucible holder with a cork stopper, add sufficient carbon fetrachloride
to cover the precipitate, cover the funnel or crucible with a watbh glass, and
allow it to stand for about half an hour. Drain off the solvent, and wash
the precipitate with further portions of carbon tetrachloride until, after
evaporating about 1 cc. of the filtrate, no visible residue remairis. Remove
the adhering carbon tetrachloride by washing the precipitate with several
portions of 10 cc. each of alcohol, and, after drying in the air, transfer to an
oven and dry to constant weight at about 110C. The weight of mercuric
sulfide, multiplied by 1.167, indicates its equivalent in HgC1 2."
The precipitate is washed with water to remove soluble sulfides,

then with alcohol te remove the water. The carbon tetrachloride, which is added later and is immiscible with water but
miscible with alcohol, can permeate the precipitate and dissolve
any sulfur formed by the decomposition of hydrogen sulfide.
The precipitate is allowed to stand covered with carbon tetrachloride for ~ hr. because sulfur is slowly soluble in carbon
tetrachloride. After washing the precipitate free of carbon
tetrachloride with alcohol, it is dried at 110C. because the
mercuric sulfide is volatile at high temperatures (400+ o C.).
47
The reactions which occur may be represented in their simplest form by the following equation:
,
HgCh
H2S~ HgS
2HCI
271.52 34.08 232.67 2(36.47)
The same principles are employed for the estimation of mercuric oxide in ointment of mercuric oxide and for the determination of the mercury content of such compounds as ammoniated
TABLE III.-:OFFICIAL SUBSTANCES ASSAYED "BY PRECIPITATION AS
SULFIDE
Substance
Sample,
ResiGm. or
due
cc.
Factor
Official requirement,
per cent
---U.S.P.
Bismuth and potassium tartrate .....
Merbaphen .........
Mercuric chloride,
poison tablets of,
large .......... , .
Mercuric chloride,
poison tablets of,
small ............
Mercuric chloride ...
Mercuric oxide, ointment of ..........
Mercury, ammoniated .............
Mercury, ammoniated, ointment of.
0.4
0.5
Bi 2S3 0.9063 Bh03 = 71 to 75

HgS 0.8622 Hg = 33 to 34.5
10
HgS
1.167 HgCl2 = 0.45 to 0.55 b
20
0.5
HgS
HgS
1.167 HgCl 2 = 0.1125 to 0.1375b

1.167 HgCl. = 99.5
10
HgS
0.931
0.5
HgS
0.8622 Hg = 78 to 80
1.5
HgS
0.862
3.0
Bi 2S. 0.9063 Bi 20 3 = 12.5 to 13.5

.
HgO = 0.9 to 1.1
Hg = 7.1 to 8.7
N.F.
Bismuth, glycerite of
Number of tablets .
Grams per tablet.
mercury and merbaphen after tQ_e mercury has been converted

into a readily ionizable salt. An analogous procedure is employed
for the estimation of bismuth compounds.
1. Show how the factor for the conversion of the weight of sulfide obtained
into an equivalent weight of mercuric chloride is derived.
48
2. How much mercuric sulfide would 1 Gm. of merbaphen containing 34

per cent of mercury yield?
3. What solvent is employed to remove the sulfur in the assay of (a)
merbaphen, (b) bismuth and potassium tartrate, (c) ammoniated mercury?
4. Explain how the presence of hydrochloric acid tends to repress the concentration of sulfide ion in the assay of mercuric chloride.
5. What special procedure is employed in the determination of mercury
in (a) ointment of ammoniated mercury and ointment of mercuric oxide,
(b) large poison tablets of mercuric chloride?
6. How would you proceed to sample (a) a ten-pound jar of ammoniated
mercury ointment, (b) a bottle containing 1,000 small poison tablets of
mercuric chloride?
7. A 5 cc. sample of glycerite of bismuth yielded 0.9094 Gm. of Bi 2S a.
Calculate the per cent of BhOa W IV in the glycerite.
Exercise 7
Determination of the Purity of Calcium Glycerophosphate.The calcium is precipitated as oxalate, ignited, and weighed as
oxide.
Object.-Assay of Calcium Glycerophosphate.
Materials Required.---O.4 Gm. of calcium glycerophosphate.
20 cc. of 5 per cent acetic acid.
Ammonium oxalate T.S. (3.5 Gm. ammonium oxalate in 100 cc.).
Procedure.-l. "Dry about 0.4 Gm. of Calcium Glycerophosphate to
constant weight at 130C., weigh accurately, dissolve in 20 cc_' of an aqueous
solution of acetic acid 1 in 20, and add 30 cc. of distilled water. Heat the
mixture to boiling, and add an excess of ammonium oxalate T.S."
Calcium glycerophosphate is hygroscopic and must be dried

at 130 to remove absorbed water. The salt is soluble in about
50 parts of water at 25, but at higher temperatures it becomes
less soluble. Acetic acid renders it more soluble, but the calcium
oxalate formed in the precipitation reaction is insoluble in acetic
acid solution. Ammonium oxalate solution is added in excess
because it reduces the solubility of calcium oxalate through
common ion effect and tends to hold any magnesium present in
solution as a double ammonium magnesium salt. Precipitation
should be carried out in boiling solution, adding the precipitating
agent slowly with constant stirring to cause the formation of
coarse crystals. The precipitate is best washed by decantation.
2. "Collect the resulting precipitate, wash, dry, and ignite to constant
weight, using a blast or high temperature burner.
49
GRAVIMETRIC METHODS
"Each gram of the residue (calcium oxide) is equivalent to 3.7473 Om. of

CaC.H.(OH).PO . '
After quantitatively transferring the precipitate to an ashless

filter and washing, the filtrate should be examined to determine
whether any crystals have passed through, since calcium oxalate
frequently comes down in fine-grained, difficultly filterable crystals. If crystals are observed in the filtrate, the latter should be
repassed through the filter. The precipitate may be ignited by
the wet method, as described on page 20. The ignited precipitate should be cooled in a desiccator and weighed as soon as
possible, since calcium oxide absorbs moisture and carbon dioxide
rapidly from the air. Upon strong ignition calcium oxalate
forms calcium oxide as shown by the following equations:
C3H~(OH)2CaP04
+ 2CHaCOOH---+Ca(CHaCOO)2 +
CaH 5 (OH)2H 2PO 4
210.15
Ca(CHaCOO)2
+ (NH4)2C20C-+CaC204 + 2CHaCOONH 4
CaC 20c-+CaO + CO 2
56.08
+ CO
Employing the above reactions as a basis, calculate the per

cent of CaO in the sample.
1. Is calcium oxalate more soluble in hot than in cold water?
2. Explain how an excess of ammonium oxalate decreases the solubility
of calcium oxalate by common ion effect.
3. Why should precipitation be carried out slowly from boiling solution?
4. Explain how to ignite a precipitate by the wet method.
5. Why should the ignition residue be weighed as rapidly as possblee?
6. How much calcium oxide should be obtained from 2.5 Gm. of pure
calcium pyrophosphate?
I
7. Briefly explain how to determine the calcium content of calcium lactate
gravimetrically.
8. Write the equations for the reactions that take place in each of the
following official assays: (a) zinc acetate, (b) manganese glycerophosphate,
(c) zinc phenolsulfonate.
9. Show how the factor in each of the assays under question 8 is derived.
50
Exercise 8
Determination of the Aluminum Content and Purity of Alulll.The aluminum is precipitated as hydroxide, ignited to oxide,
and weighed.
Object.-Assay of Alum.
Materials Required.-l Gm. of alum.
1 Gm. of ammonium chloride.
Ammonia T.S.
Procedure.-" Dissolve about 1 Gm. of Alum, accurately weighed, and
about 1 Gm. of ammonium chloride in 250 cc. of distilled water. Heat the
solution to boiling, and Mid a slight excess of ammonia T.S. to precipitate
aluminum hydroxide. Collect the precipitate on a filter, wash thoroughly
with hot distilled water, dry, ignite strongly, and weigh. The weight of
the aluminum oxide so obtained, multiplied by 8.894, indicates its equivalent
in Al'NH.(S04)2.12H 20 and when multiplied by 9.307, indicates its equivalent in AlK(SO.):.12H 20."
Ammonium chloride is added to the dissolved alum to prevent

the formation of the colloidal form of aluminum hydroxide and
to prevent the precipitation of other hydroxides of metals,
as magnesium, which may be present,as- impurity. Ammonia
water is added to precipitate the aluminum as hydroxide. The
ammonia water should be freshly distilled, since upon standing
in glass containers it dissolves silica, the presence of which leads
to high re~mlts, the silica being precipitated and retained in the
precipitate. The ammonia water should be added in very slight
excess, since aluminum hydroxide is slightly soluble in strong
solutions of ammonia. The precipitation is conducted in boiling
solution to convert any colloidal aluminum hydroxide into large
particles and to secure a coarse-grained precipitate. The solution
should not be boiled after complete precipitation, for ammonium
salts become acid in reaction upon prolonged boiling due to loss
of ammonia, and the resulting acid solution would dissolve some
aluminum hydroxide. Aluminum hydroxide forms a slimy
precipitate difficult to wash. It is most easily washed by decantation, using a hot wash liquid prepared by adding a drop of
ammonium hydroxide ~o hot distilled water. As much of the
supernatant liquid should be passed through the filter as possible
after the third washing before transferring the precipitate to the
filter, since the nature of the precipitate renders filtration very
51
slow. The precipitate ignited by the wet method should be

heated strongly, preferably over a blast lamp, after the ash has
T~IiE IV.-OFFICIAL SUBSTANCES ASSAYED BY IGNITION TO OxIDE
Substance
Sample,
FacOm. or Residue
tor
cc.
Official requirement, per cent
- - - --U.S.P.
Alum, ammoniUDl .......
Alum, ammoniuOl, exsiccated ...............
Alum, potassium ........
Alum, potassium, exsic...
cated ................
Bismuth subcarbonate ...
Bismuth subgallate ......
Bismuth subnitrate .....
Bismuth subsalioylate ..
Calcium creosotate ......
Zinc ac.tate ............
Zinc chloride ............
Zinc oxide, ointment of ..
Zinc suifate .............
N.F.
Aluminum acetate, solution of ...............
Aluminum chloride ......
Aluminum chloride, solution of. ..............
Aluminnm subacetate,
solution of ............
Aluminum sulfate .......
Bismuth magna ........
Bismllth subcarbona.te,
tablets of .............
Bismuth SUbgallate, tablets of
Bismuth subnitrate, tablets of ................
Bismuth subsalicylate.
ampuls of .............
Calcium glycerophosphate
Manganese citrate. soluble
Manganese glycerophosphate ................
Zinc phenolsulfonate ...
1.0
AhO.
8.894
AINH.(SO.),.12H,O = 99.5
0.5
1.0
AhO.
AhO.
4.653
9.307
AINH.(SO.), '" 96.5

AIK(SO,),.12IhO - 99.5
0.5
1.0
1.0
1.0
1.0
0.2
1.0
1.0
2.0
1.0
AhO.
BisO.
Bi,O.
BitO.
BhO.
CaO
ZnO
ZnO
ZnO
ZnO
5.066
AIK(SO.), = 96.5
BhO. = 90
Bi,O. = 52 to 57
.BhO, - 79
BhO, - 62 to 66
CaO = 40 to 50
(CH,COO),Zn ~ 83.16 to 87.32
ZnCh - 95
ZnO = 19 to 21
ZnSO. = 55.86 to 58.63
5.0
0.5
AhO.
AhO.
4.0031 AI(C.H,O.,. = 4.8 to 5.8 W IV

4.7369 AICIa.6H,O = 95
5.0
AhO.
4.737
5.0
0.5
10.0
AbO.
Al,O,
BilO.
3.1788 Al(C,HaO,hOR = 7.5 to 8.5 W IV

6.537 Alo(S04) . 18H,0 = 99.5
...... Bi,O. = 5.6 to 6.2WIV
3.0-
Bi,O.
...... Bi,O.
= 83 to 97 b
a.o
Bi,O.
...... Bi,O.
= 48 to 6]0
3.0
BilO.
......
1.0
0.4
0.5
BisO. ...... BhO, - 57.6 to 70.4'

3.7473 CaC,H.(OH),PO. -= 98
CaO
Mn.O. 2.3728 [C.H . OR(COO).],Mn. = 48 to 52
0.5
2.0
Mn.O, 2.95
MnC,H.(OHhPO. = 98
ZnO
6.8284 Zn(C,H.OSO.) . 8H,O = 99.5
......
......
......
......
......
2.254
1.674
......
1.984
AlCla.6H,O = 22.5 to 27.5WIV
BhO. = 73 to 85'
Weight of ingredient sought.

Per cent of labeled amount.
become white to convert any traces of basic aluminum sulfate

to the oxide:
52
2AINH.(SO')z.12HzO
2 X 453.32
+ 6NH.OH--i>2AI(OH)3 + 4(NH.)zSO. + 24H zO

2 X 77.99
2AI(OH).__"Al zO a + 3H zO.
101.94
906.64:101.94: :x:1
101.94x = 906.64
x = 8.894 Gm. alum equivalent to 1 Gm. AlzO a
Calculate the purity of the alum and the per cent of aluminum
in the sample.
1. Why is ammonium chloride added to the alum solution previous to
precipitation?
2. Why should a large excess of ammonium hydroxide be avoided?
3. Indicate by reactions how ammonium salts in solution may become
acid in reaction upon prolonged boiling.
4. If an unknown sample of alum yields 0.125 Gm. of Al z0 3 upon assay, to
how much AINH.(SO.)z.l2HzO, AINH.(SO.)" and AIK (SO.)z. 12H zO,
respectively, is the Al 20, equivalent?
6. A 3.0000 Gm. sample prepared by powdering 20 tablets of bismuth
subnitrate which weighed 6.8420 Gm. yielded 2.3687 Gm. of Bi z0 3 Calculate the amount of Bi and of BizO. contained in each tablet. Calculate the
per cent of Bi and of Bi z0 3 contained in each tablet.
6. Write equations for the reactions that take place in the assay of ointment of zinc oxide.
7. How much pure zinc sulfate would be required to form 0.8920 Gm. of
ZnO?
8. A sample of soluble manganese citrate weighing 0.5624 Gm. yielded
0.2744 Gm. of Mn 30.. What per cent of manganese did the sample contain?
Exercise 9
Determination of the Magnesium in a Magnesium Salt.-The

magnesium is precipitated as magnesium ammonium phosphate,
washed, dried, ignited to pyrophosphate, and weighed.
Object.-Assay of Solution of Magnesium Citrate for Magnesium Oxide.
Materials Required.-lO ee. of solution of magnesium citrate.
20 ce. of sodium phosphate T.S. (12 Gm. NazHPO. in 100 cc. of distilled
water).
Stronger ammonia T.S. (at least 27 per cent NH.).
Ammonia T.S. (9 to 10 per cent NH.).
Procedure.-l. "Transfer to a beaker of about 200-cc. capacity exactly
10 ee. of Solution of Magnesium Citrate which has been previously freed
GRAVIMETRIC METHODS
53
from excessive carbon dioxide by repeatcd pouring. Add 100 cc. of distilled water, 2 cc. of hydrochloric acid, 20 cc. of sodium phosphate T.S.,
and 2 drops of methyl red T.S. Add ammonia T.S a few drops at a time
with constant stirring until the solution becomes faintly yellow. Allow the
mixture to stand for ten minutes, add 40 cc. of stronger ammonia T.S. with
constant stirring, and allow the mixture to stand for two hours or over night."
The sample is' measured after removing most of the carbon

dioxide gas by pouring to prevent loss due to effervescence.
When sodium phosphate is dissolved in water, the phosphoric
acid ionizes in three stages which may be represented by the
following equationl'l:
H+
H aP0 4
H 2P04"<- 'H+
HPO'4
H+
+ H 2P04"
+ HPO,
+ PO:'
The degree of ionization becomes less for each successive stage.

The relative amount of PO:" HPO'4, H 2P04", and H aP0 4 in
equilibrium with each other depends upon the hydrogen ion
concentration of the solution. In a basic solution, the PO~ and
HPO, ions predominate. Each of the above species of phosphate
ion which exist in the solution of phosphate reacts with Mg++
ions to form the corresponding magnesium salts, all of which
are more or less insoluble in water. Thus there may be formed
Mg a(P0 4)2, MgHP0 4, and Mg(H 2P0 4h The addition of a
solution of NH 40H represses the concentration of H 2P04" ions
and prevents the formation of Mg(H2P04h. In the presence
of the ammonium ion, the salt is precipitated as MgNH 4P04.6H 20. If the solution is too basic, Mg(OH)2 may be precipitated. The precipitation of both (Mg aP0 4)a and Mg(OHh is
prevented by the presence of ammonium salts which, through
common ion effect, act as a buffer and prevent the OH ion
concentration from becoming too great. The hexahydrate,
MgNH 4P0 4 .6H 2 0, forms in relatively coarse crystals on standing
for three or more hours at room tem:peratures. At temperatures
above 60C. the monohydrate, MgNH 4 P0 4 .H zO, is formed.
2. "Collect the precipitate in a weighed Gooch crucible and wash with a
mixture of 1 volume of ammonia T.S. and 3 volumes of distilled water until

free from chloride. Dry and ignite to a constant weight. The weight of
magnesium pyrophosphate (Mg,P,O,) so obtained when multiplied by 3.621
gives the amount of magnesium oxide represented by 100 cc. of the solution."
54
The precipitate is washed with aqueous ammonia solution

until free of chlorides. In testing the filtrate for chlorides,
acidulate with nitric acid before adding the silver nitrate solution. Upon ignition, the magnesium ammonium phosphate is
converted into the pyrophosphate as follows:
2MgNH4P04.6H20~Mg2P207
+ 2NHa + 13H 20
For the purpose of making calculations the equation may be

represented as
2MgO~Mg2P207
2(40.32)
222.68
Thus it is evident that each gram of Mg 2P 20 7 is equivalent to

80.64/222.68 = 0.3621 Om. of MgO, and, since 10 cc. of the
TABLE V.-OFFICIAL SUBSTANCES ASSAYED GRAVIMETRICALLY AS
PYROPHOSPHATE
Substance
Sample,
Gm. or
co.
Official requirements,
per cent
Residue Factor
--
U.S.P.
Magnesium citrate, solution
of ............
Magnesium sulfate ..... " ...
Sodium phosphate .........
Sodium phosphate, exsiccated .........
N.F.
Magnesium sulfate, ampuls of
Sodium phosphate, solution
of ............
10.0
Mg2P 2Or 0.3621 MgO
= 1.6 to
1.9W IV
= 99.5
1.0
Mg~20r
0.3
Mg 2P 2Or 1.275
Na.HPO.
= 98
0.2
Mg 2P 2Or 1.275
Na,HPO.
= 98
0.20
Mg 2P 2Or 2.2138 MgSO.7H 20 = 95 to
5.0
Mg 2P 2Or 1.275
1.081
MgSO.
Na,2HPO.
= 39 to
105~
41 W IV
Weight of ingredient Bought.

b Per cent of labeled amount.
solution was used as sample, 10 X 0.3621 = 3.621 Om. of MgO

in each 100 cc. of solution.
55
The principles involved in this exercise may be applied to the

analysis for magnesium in all soluble magnesium salts. Conversely, phosphates may be determined by essentially the same
method, magnesia mixture being added to the solution containing
phosphate ion. The arsenate ion, AsO~, may be precipitated in
like manner to yield MgNH 4As0 4 Salts similar to MgNH 4P0 4
TABLE Vr.-SOME OTHER OFFICIAL GRAVIMETRIC ASSAYS
Substance
Sample,
Gm. or
Residue
Official requirement. per cent
ee.
U.S.P.
Collodion ............. 10.0
Camphor, liniment of ..
5.0
Camphor. spirit of. .... 25.0
Erythrityl tetranitrate,
diluted.
Gold chloride, R ......
Iodine, compound, 80lution of (for KI) ......
Iodine, tincture of (for
KI) ...................
Iodine. tincture of, mild
(for NaI) ...........
Molybdic anhydride, It.
Palladous chloride, R ..
Platinic chloride, R ....
N.F.
Camphor, ampule of ...
Phenolphthalein, tahlets
01 ..................
Pyroxylin ~ 5.1 W IV
Camphor ~ 19 to 21
Camphor = 9.5 to 10 5 W IV
0.2
Pyroxylin
Cottonseed oil
Camphor dinitrophenylhydrazide
Erythrityl tetranitrate
Au
Au
= 47
5.0
KI
KI
= 9.5
5.0
KI
KI
= 4.5 to 5.5WIV
5.0
0.5
0.2
0.5
Nal
PbMoO.
Pd
Pt
NaI = 2.1 to 2.5W IV

MoO, = 99.5
Pd = 59
Pt = 37
5.0
Ampul oil
Camphor
0.65"
Phenolphthalein
Phenolphthalein = 92.5 to 107.5&
0.25
C.H,(NO.)<
47 to 53
to 10.5 W IV
....
= 93 to 103&
" Amount equivalent to .

Per cent of the labeled amount.
R. - reagent.
are formed by zinc, cadmium, and manganese. Under carefully controlled conditions, the same general principles may be
made the basis for the estimation of cadmium, manganese,
phosphorus, zinc, and arsenic.
~uestions
and Problems
1. Explain each step in the procedure for the assay of sodium phosphate
and write equations for all reactions that take place in the assay.
2. What is magnesia mixture T.S.?
8. Under what conditions is Mg++ precipitated as MgNH 4P0 4.2H zO?
56
4. In the assay of ampuls of magnesium sulfate labeled "each 2 cc. contains 1 Gm. of magnesium sulfate," how would you sample a shipment containing 1 gross of ampuls? How much of the ampul solution would you
measure for each assay sample? What minimum and maJ[imum amounts of
magnesium sulfate might the ampuls contain in each cubic centimeter and
meet the official tolerance requirements?
5. A sample of sodium phosphate weighing 0.3666 GIn., when dried to
constant weight at 110C., weighed 0.1942 Gm. and upon assay yielded
0.1491 Gm. of Mg 2P 207. Calculate the per cent of sodium phosphate and
of phosphorus in the original sample and in the moiBturefree sample.
What per cent of moisture did the original sample contain?
Other Gravimetric Assays.-Numerous gravirnetric methods

of assay other than those considered in this chapter are used in
the determination of official substances. Those which require
a special technique, such as the gravimetric asslty of alkaloids,
are treated in subsequent chapters. Some of the official gravimetric assays, such as the ones for alkali iodides, which involve
only evaporation, drying to constant weight, and weighing, are
indicated in the table on page 55.
CHAPTER IV
PRINCIPLES OF VOLUMETRIC (TITRIMETRIC)
ANALYSIS
In volumetric (titrimetric) analysis, or analysis by measure,

the quantity of an element, component of a compound, or compound in a weighed sample is made to react with a measured
volume of a reagent solution of known concentration. Since the
volume and concentration of the reagent solution required to
complete a definite reaction are known, the amount of substance
entering into reaction with the reagent solution can be calculated.
Thus the chloride ion content in a soluble chloride is determined
by dissolving the chloride in water and adding silver nitrate
solution of known concentration until practically all of the chloride ion has been precipitated as silver chloride. From the volume
of silver nitrate solution consumed, the weight of chlorine in the
sample and the purity of the soluble chloride can be calculated.
Definitions.-The operation whereby the concentration or
value of a solution for a specific reaction is determined is termed
standardization. For example, a known weight of pure sodium
carbonate dissolved in water, when made to react quantitatively
with a measured volume of hydrochloric acid, gives the data
required to calculate the concentration of hydrochloric acid in
the solution. The hydrochloric acid solution thus standardized
is called a standard solution, i.e., a solution which contains a
known weight of reagent per unit of volume. The process of adding a standard solution or reagent in measured quantity to a
substance until the end point of reaction is observed is called
titration. Consequently, volumetric methods are often synonymously termed titrimetric melhods. Titration may be conducted
directly, i.e., a standard acid is added to an alkali until the end
point is reached, or residually, i.e., an excess of standard acid is
added to the unknown alkali and the amount of excess acid is
determined by titration with standard alkali solution. The end
point is the practical point at which titration is stopped when
57
58
the reaction is complete as shown by a particular indicator or

other device used for that purpose. The end point should not
be confused with the stoichiometric point, i.e., the point which
marks the addition of the calculated equivalent of reacting substance demanded by the equation.
Volumetric Apparatus.-The apparatus required for v'olumetric
analysis includes the analytical balance described under Chapter
I and a number of accurately calibrated instruments as well
as beakers, flasks, funnels, etc. Volumetric apparatus is of two
types: that made to deliver a definite volume of liquid, as burettes
and pipettes; and that made to contain a definite volume of
liquid, as volumetric flasks and graduated cylinders. The temperature specified in the U.S.P. for volumetric measurements
is 25C., while the Bureau of Standards has adopted 20C. as the
standard temperature for glass volumetric apparatus. It matters little at what temperature the instruments are calibrated
provided the same temperature is used for all, and all measurements are made at the temperature for which the instruments are
calibrated.
Units of Capacity.-The U.S.P. liter is the volume occupied by
996.04 Gm. of distilled water at 25C., weighed in ~ir with brass
weights, with the barometric pressure at 760 mm. The official
cubic centimeter (cc.) represents the one-thousandth part of the
normal liter. The normal liter is defined as the volume occupied
by a kilogram of water weighed in vacuum at 4C. The onethousandth part of a normal liter, termed a milliliter, and the
cubic centimeter, are both used as the unit of capacity. The cubic
centimeter, abbreviated as "cc.," which has been adopted as the
official unit of capacity, is not exactly the one-thousandth part of
a normal liter, since one milliliter = 1.000027 cubic centimeters.
The difference is sO'slight, however, that it becomes negligible in
all volumetric measurements.
Burettes.-Graduated glass tub'es of uniform bore throughout
the whole length used in the measurement of variable quantities'
ofliquid are termed burettes (Figs. 9 and 10). They are graduated into cubic centimeters and closed at the bottom by a glass
stopcock or a short soft india rubber tube provided with a glass
tip and pinchcock or other suitable device to control the outflow
of liquid. The official requirements for burettes follow:
PRINCIPLES OF VOLUMETRIC ANALYSIS
59
"All burettes should be provided with glass stop-cocks and

calibrated for use at the standard or other temperature at which
they are to be used.' Burettes provided with rubber tubes,
pinch cocks and glass delivery tubes in place of glass stop-cocks
FIG. 9.-Burette with glass

stopcock.
FIG. lO.-Burette wi1

pinchcock. (Mohr burette.)
may be used for potassium and sodium hydroxide solutions and

such other solutions as are best used in this type of burette. The
glass tips should be from 2 to 3 cm. in length, gradually tapered
and slightly bent. The rate of outflow should be regul!J.ted so
that not less than two seconds should be consumed for each cc.
delivered. When completing a titration, the tip of the delivery
tube should be touched to the wet inner surface of the receiving
vessel and the solution again stirred."
Burettes provided with glass stopcocks should be tested for
leakage. A lubricant such as petrolatum serves to prevent leak-
60
age and permits easy operation, but an excess of lubricant should

never be used. Those provided with a rubber outlet (Mohr
burettes) should never be used for other than alkali solutions.
The outlet tip of either type of burette should be of such diameter
and taper as to permit the delivery of a drop the volume of which
is considerably less than that which can be accommodated
between the finest graduations of the scale with
which the burette is marked.
Reading Burettes.-The surface of liquids in
narrow tubes 1S always curved due to capillarity j
the surface or meniscus is always concave when
the liquid wets the tube and convex when it does
not. Readings should always be made at the lowest
point of the meniscus, that is, the graduation mark
which coincides with the bottom of the curve, except
when measuring highly colored liquids. The eye
must be on the same level as the meniscus, otherwise error will be introduced due to parallax (see Fig.
~F~il~
11). Numerous devices have been employed to
Effect of
assist
in reading burettes correctly. One such device
parallax in
reading bu- recommended by the United States Bureau of
rettes.
Standards is a collar-shaped section of black rubber
tubing, cut open at one side and of a size which fits the burette
tube so that it can be moved. This black rubber tubing, when
placed immediately below the meniscus, renders the meniscus
profile dark and clearly observable against a light background.
Pipettes.-The following requirements for pipettes have been
adopted in the U.S.P.:
"Pipettes should be graduated to deliver at standard temperature the volume indicated. The suction stem should be at
least 16 cm. long,. and the delivery tube not less than 3 cm. and
not more than 25 cm. long. The inside diameter at the capacity
mark must not be less than 2 mm. It must not exceed 4 mm.
for pipettes up to and including 25-cc. capacity, 5 mm. for 50-cc.
capacity and 6 mm. for 100-cc. and 200-cc. capacities. The
capacity mark must not be more than 6 cm. from the bulb. The
outlet of any transfer pipette must be of such size that the free
outflow for water shall last not more than one minute. Not less
than fifteen seconds shall be required to empty a 5-cc. pipette,
61
twenty seconds for a 1O-cc. pipette, thirty seconds for a 50-cc.

pipette, forty seconds for a 100-cc. pipette, and a minimum of
fifty seconds for a 200-cc. pipette. After filling, the liquid adhering to the outside should be wiped from the stem. When emptying the contents, the pipette should be held in a vertical position
and the outflow should be unrestricted until the surface of the
water reaches the upper end of the delivery tube; the tip should
then be touched to the wet surface of the receiving vessel and kept
in contact with it until the emptying is complete. Pipettes
should never be drained by blowing into them unless, as in the
case of the Ostwald pipettes, they are especially graduated for
use in this way.
"The limits of error permitted in the calibration of pipettes
shall be those accepted by the United States Bureau of Standards,
which are as follows:
Contents of pipettes
Limit of error in cc.
Limit of error in per cent
2 cc.
0.006
0.30
5 cc.
0.01
0.20
10 cc.
0.02
0.20
25 ce.
0.03
0.12
50 cc. 100 cc.

0.05
0.08
0.01
0.08
"It is essential that ip. using pipettes the same procedure be

employed as was used in the calibration."
In using transfer or delivery pipettes (Fig. 12), they are filled
from the jet to about 1 cm. above the mark, then allowed to run
down just to the mark. The liquid is drawn up by sucking the
upper end of the pipette with the mouth, but this procedure
should be, avoided and some other method of measurement
employed when volatile, corrosive, or poisonous liquids are
measured.
Measuring Flasks.-The U.S.P. requirements for measuring
or volumetric flasks (Fig. 13) are as follows:
"Standard measuring flasks are calibrated to contain, when
filled to the mark, 1000,500, 258, 200, 100 or 50 cc. at 25C. The
necks must measure not less, than 14 mm. and not more than
20 mm. in diameter for 1000-cc. capacity; from 12 mm. to 18 mm.
for 500-capacity; from 10 mm. to 15 mm. for 250-cc. capacity;
from 8 mm. to 12 mm. for 100-cc. capacity, and from 6 mm. to
10 mm. for 50-cc. capacity. The capacity mark on any of these
flasks should be not less than 6 cm. distant from the mouth and
not less than 2 cm. from the base of the neck. The limits of
62
error permitted in the calibration of flasks shall be those accepted

by the United States Bureau of Standards, which are as follows:"
Contents of flask
Limit of error in ee.
Limit of error in per cent
50 ee.
0.05
0.10
100 ee. 250 ee.

0.08
0.12
0.08
0.05
500 ee. 1000 ee.

0.15
0.30
0.03
0.03
Volumetric flasks are used to make up standard solutions to a

given volume. Note that the U.S.P. requires that flasks be
graduated to contain and not to deliver the calibrated
volume.
Graduated Cylinders.-Graduated cylinders are
used in making approximate measures of volume.
They are graduated to contain a given volume of
liquid at standard temperature. The inside diameter of the cylinder should not exceed one-fifth of the
graduated length.
Cleaning Volumetric Apparatus.-New volumetric apparatus should always be cleaned before using,
and that in use should be cleaned as often as necessary. The adherence of droplets to the wall of a
burette or pipette is positive. evidence that the apparatus is dirty. In a clean instrument, the liquid
drains
down uniformly wetting the
a
walls so that no droplets are observable. A warm cleaning solution (sodium dichromate in sulfuric acid),
solution of sodium hydroxide, or solution of soap powder is the best cleansing agent to use. Hot solutions
should be avoided when cleaning accurately calibrated apparatus because of
the possib}e production of a permanent
change in volume caused by the heat
FIG, 12.- and known as "thermal after-effect."
(a) Transfer
Calibration of Volumetric Apparatus.
or delivery
pipette, (b) The ordinary volumetric apparatus
FIG. 13. cal'b
Measuring
d at varIous
.
Volumetric
mar
ete
IS
1
rate
temflask.
k
d
pipette.
peratures and is often unreliable. The
analyst should always check the calibration of volumetric apparatus and make corrections if necessary.
63
Burettes.-The calibration of burettes may be performed in

various ways by means of special calibration devices such as the
Ostwald calibrating pipette and the Kiehl burette calibrator. In
all practical work, however, the following method is satisfactory:
The clean burette previously tested for leakage is filled to the
zero mark with distilled water. The burette and water used
should be in the room a sufficient length of time before use to
acquire room temperature. A definite volume, e.g., 10 cc., is
allowed to run into a tared beaker. The weight of water withdrawn is determined and the burette is filled again to the zero
mark and then 20 cc. is withdrawn in exactly the same way.
This operation is repeated for each 10 cc. until the 50 cc. mark is
reached, determining the weight of water withdrawn each time.
The results with corrections should be recorded in tabular form
showing the relation between the apparent volume and actual
weight delivered to each mark on the burette corresponding to an
interval of 10 cc. Weighings need not be more ~ccurate than to
the second decimal place, since that is the greatest degree of
accuracy with -which the burette can be read. In practical
calibration, 1 cc. may be considered equal to 1 Gm.
A burette calibrated at 20C. should deliver 9.9718 Gm. of
water or at 25C. it should deliver 9.9604 Gm. for each 10 cc.,
since 1 liter of distilled water at 20C. weighs 997.177 Gm. and at
25C., 996.04 Gm. when weighed with brass weights in air of 50
per cent humidity at standard atmospheric pressure. The true
volume for each 10 cc. segment of the burette may be calculated
from the weights obtained and recorded on a convenient chart.
Pipettes.-Clean the pipette thoroughly with soap solution
and cleaning mixture and rinse well with distilled water. Fill it
with distilled water at 20 or 25C. according to the temperature
selected for calibration, and allow it to drain into It' tared flask.
Weigh accurately and record the correct volume of the
pipette.
Flasks.-Clean, dry, and tare the flask, placing the number of
grams weight corresponding to its volume on the opposite balance
pan and introduce water at 20 or 25C. into the flask in sufficient
quantity to counterbalance the added weights. Mark the lowest
point of the meniscus if it does not agree with the original
calibration.
64
Sources of Error in the Use of Volumetric Apparatus.-Every

precaution should be taken to avoid the following common
sources of error in the use of volumetric apparatus:
1. Rinse water adhering to walls of apparatus. The apparatus
may be dried, but it is more convenient and just as accurate to
wash it out with small successive portions of the liquid to be
used, discarding the washings.
2. Grease films and dirty apparatus cause irregularity in the
delivery of liquids and distort the meniscus.
3. Drainage or afterflow of the liquid adhering to the vessel
walls to the liquid below necessitates that the time of drainage
be the same as that used in the calibration of the instrument.
4. Parallax must be avoided to secure proper readings of the
level 9f the meniscus.
5. Variations in temperature lead to changes in volume of
vessels and liquids. All measurements, therefore, should be
made at a temperature closely approximating that at which the
apparatus was calibrated.
6. Air bubbles trapped beneath the liquid surface, especially
below the stopcock in burettes, displace liquid.
7. Heat, such as supplied by hot solutions, causes calibrated volumetric apparatus to suffer a slight permanent chanpe of volume.
S. Most salts when dissolved produce a change in temperature
with concomitant change in volume of the solution. Such solutions should never be measured until they have acquired the
temperature at which the instruments used to measure them were
calibrated.
9. Failure to use apparatus in a manner approaching as nearly
as possible that followed in calibration.
1. Define the followig terms: (a) standardization, (b) standard solution,

(c) end point, (d) residual titration.
2. What is the official liter?
3. Burettes provided with rubber tubes and pinchcocks in place of glass
stopcocks may be used to measure out sodium or potassium hydroxide
solution, but they should never be used in the measurement of solutions of
acids or oxidizing agents. Why?
4. A 100 cc. volumetric flask calibrated at 25C. is used to measure water
at 20C. What correction of the volume must be made? (See U.S.P. Xl
table, page 623.
65
5. A volumetric flask calibrated to contain 100 cc. at 25C. and 760 mm.
barometric pressure is used at 18C. and 746 mm. barometric pressure.
Calculate the volume of water contained in the flask at the temperature
and pressure at which it is used.
NEUTRALIZATION METHODS-ACIDIMETRY AND ALKALIMETRY
Acidimetry is the measurement of the quantity of acid in a

given sample by titration with a suitable alkali. Alkalimetry
is the measurement of the quantity of alkali in a given sample by
titration with a suitable acid. In each case the principle involved
is the same, namely, to add to the acid o~ alkali being determined
an equivalent amount of standard solution of alkali or acid,
respectively. From the amount of standard solution added, the
amount of alkali or acid originally present is calculated by means
of the stoichiometric equation. The point at which an equivalent
of reacting substance has been added to another substance is
termed the stoichiometric point, equivalence point, or theoretical
end point. It is that point at which equivalent amounts of reacting
substances have been combined as demanded by the equation. In
practice the titration is stopped at a practical point, termed the
end point, when the reaction is shown to be complete by the
change in color of a particular indicator or by some other device.
In every titration the end point and stoichiometric point must
coincide very closely if the result is to be of value in analytical
work.
Before undertaking analyses by methods of acidimetry and
alkalimetry, the student should become familiar with the theoretical considerations involved.
Titer.-The. titer of a solution is the number of grams of solute
contained in 1 cc., or the weight of any substance which will
react with or be exactly equivalent to 1 cc. of the solution, e.g.,
a solution containing 36.47 Gm. of HCl in 1,000 cc. has an HCl
titer of 0.03647. The NaOH titer of this solution may be found
\.,
from the relationship:
HCl:NaOH::0.03647:x
where x is the weight in grams of pure NaOH which can be
neutralized by 1 cc. of the' above acid and is equal to 0.0400 Gm.
66
In like manner, the desired titer of any other substance may be

found by direct proportion.
Theory.-The hydrogen ion concentration influences most
reactions employed in analytical methods whether they are
neutralization, precipitation, or oxidation-reduction reactions.
It has been established that in any aqueous solution there are
always hydrogen and hydroxyl ions present and that whatever
the concentration of the one, the concentration of the other
is such that the product of their concentrations is a constant for
any given temperature.
[H+] X [OH-] = a constant for any given temperature, and
in pure water at about 220., [H+] X [OH-] = 1 X 10- 14 Since
this relationship is true, the concentration of hydrogen ions [H+]
expressed in terms of gram-ions per liter in pure water is equal
to the square root of 1 X 10- 14 = 1 X 10-7 (0.0000001), and the
hydroxyl ion concentration [OH-] also is equal to 1 X 10-7
The degree of acidity or alkalinity of a solution can, therefore,
be defined in terms of its concentration of hydrogen ion or
hydroxyl ion, since when the value of one is known the value of
the other can be calculated; i.e., if [H+] = 10-5, then [OH-] =
10-9 , since [H+] X [OH-] must equal 10- 14 Solutions which
7
have a hydrogen ion concentration greater than
are acidic,
and those that have a hydrogen ion concentration less than 10-7
are alkaline. Thus a solution of 0.01 N HOI contains 10-2
Gm.-ions of hydrogen per liter, assuming complete dissociation,
and is acid in reaction although it contains 10- 12 Gm.-ions of OH-.
To avoid the disadvantage of employing a dual system of
definition, it is customary to use only the hydrogen ion concentration to characterize a solution, whether it be acidic, neutral, or
alkaline, the hydroxyl ion concentration being implied.
pH Value.-In addition to the exponential form of expressing
hydrogen ion concentration employed above, another system
suggested by Sorensen is in common use. In the latter system,
only the exponent is used with its negative sign dropped and thy
symbol pH prefixed, e.g.:
\0-
[H+] = 10-2 . 06 then pH = 2.06 acidic

[H+] = 10-7 then pH = 7 neutral
[H+] = 10-12 . 95 then pH = '12.95 alkaline
67
pH is generally defined as the logarithm of the reciprocal of the

hydrogen ion concentration, written pH = log [J+j'
Since the
hydrogen ion concentration of a solution is a fraction denoting

the number of grams of hydrogen ion contained in a liter, it can
be expressed 'as l/[H+] which is the reciprocal of [H+]. By
the use of the reciprocal, the negative exponent is avoided; thus,
1/104 is the same as 1 X 10-4
The fact that a solution contains 0.0001 Gm. of hydrogen ions
per liter may, therefore, be expressed in various ways as follows:
[H+]
[H+]
log [H+]
- log [H+]
1
log [H+]
pH
= 0.0001 Gm. per liter

= 1 X 10-4
= - 4
= 4
= 4
=4
Concentrations that are uneven decimal fractions can also be

expressed in terms of pH units; e.g., if the hydrogen ion concentration of a solution is 2.73 X 10-4 the pH value is 3.564. By
reference to a logarithm table, it is found that log 2.73 = + 0.436
and log 10-4 = - 4.000. .Since the numbers are to be multiplied,
the logs are added and -4.000 + 0.436 = - 3.564. Therefore
log [H+] = -3.564, -log [H+] = 3.564, and pH = 3.564.
Conversely, when the pH is given, the hydrogen ion concentration can be calculated. Thus pH = 9.63 means that [H+] =
1 X 10-9 63 The exponent -9.63 = -10 + 0.37, and 10-9 . 63
= 10-10 X 10+ 0 . 37 From the logarithm table, it is found
that the exponent 0.37 corresponds to the number .2.34. Therefore [H+] = 10-9 63 is equal to [H+] = 2.34 X 10- 10
Two practical methods of determining when the hydrogen
ion concentration'in a solutiop. has acquired a certain value have
been developed, namely, the potentiometric method, and the
colorimetric method (see page 272).
Buffers.-In many analytical procedures a change in the
hydrogen ion concentration occurs as a result of chemical reaction. In certain cases, this change in the concentration of hydrogen ion may prevent the reaction from going to completion
68
quantitatively. Since the change in hydrogen ion concentration

is caused by the formation of acid or base in the course of ,the
reaction, some provision must be made to eliminate the acid or
base formed in such a manner as to maintain a sensibly constant
hydrogen ion concentration. This is usually accomplished by
means of a buffer. The term buffer is applied alike to the salts
of weak acids and to the salts of weak bases. Solutions of these
salts resist change in hydrogen ion concentration on the addition
of small amounts of either Mid or base. In the titration of
acetic acid with sodium hydroxide, the sodium acetate formed
acts as a buffer to prevent 11 large change in hydrogen ion concentration. Acetic acid in aqueous solution is slightly dissociated
as shown in the following equations:
(1)
CRaCOOR
NaOR
+ CRaCOO+
OR- + Na+
R+
(2)
11(3)
ROR
11(4)
+ CRaCOONa
In reaction (1), the slightly dissociated acetic acid forms R+

and CRaCOO- ions which combine with the O;H- and Na+
ions from equation (2) to form ROR in equation (3) and CRsCOONa in equation (4). The sodium acetate is highly dissociated into Na+ and CRsCOO- ions. This reslilts in a high
concentration of CHsCOO- ions in the solution and repression
of the R+ ion concentration' forcing equation (1) toward the left.
As additional quantities of NaOR are added, the R+ ion is
removed as ROR and further small amounts of acetic acid dissociate so that a continuous supply of R+ ion is provided. The
solution will continue to contain an excess of R+ ion until all of
the CHsCOOR has been consumed. The addition of a small
quantity of alkali in excess of the required amount will then
cause the OR- ion concentration to exceed the R+ ion concentration and the solution will react alkaline.
The principle of buffer action is often employed in analytical
methods in which acid or base is generated as a result of the
analytical reaction. In such cases, the addition of an amount of
buffer salt relatively large with respect to the amount of acid or
69
base generated causes a fractional diminution of the buffer ion,

through its union with the hydrogen ion to form the slightly
dissociated acid corresponding to the buffer salt employed. The
diminution in the amount of buffer salt will then be so slight that
the hydrogen ion concentration will remain practically constant.
In iodimetric assays where HI is formed as a product of the reaction, sodium bicarbonate is used as a buffer to maintain an
approximately constant hydrogen ion concentration.
Neutralization Reactions.-It is important to remember that,
in terms of the ionic theory, a neutral solution contains H+ and
OH- ions in the same concentration as in pure water, namely,
0.0000001 Gm.-ion per liter of solution. Any substance which
can furnish excess H+ ions when dissolved in water is acidic and
any substance which can furnish excess OH- ions when dissolved
in water ,is basic. From the point of view of the ionic theory,
therefore, neutralization is the union of H+ ions from an acid with
OH- ions from a base, to form molecules of water. The other
ions furnished by the acid or base mayor may not remain
combined.
Neutralization reactions must proceed to completion to be of
value in quantitative analysis. Reactions may be caused to go
to completion in a number of ways, i.e., by the formation of a
slightly dissociated substance as a reaction product; by the
removal of one or more of the products of the reaction as a gas;
by the removal of one or more of the products of reaction as a
precipitate; and by adding an excess of one of the reactants.
Thus, if HCI and NaOH are dissolved separately in water to
make dilute aqueous solutions, each substance is almost completely dissociated into ions [reactions (1) and (2)].
(1)
HCI
NaOH
+ CI+
OH- + Na+
H+
(2)
~ il(3)
HOH
11(4)
NaCI
Upon mixing the two solutions some Na+ and CI- ions unite
forming NaCl molecules-reaction (4). Water is only slightly
70
ionized, so nearly all of the H+ ions and OH- ions unite to form
HOH molecules-reaction (3). The removal of H+ and OHions in reaction (3) causes reactions (1) and (2) to proceed to
completion, so that if equivalent quantities of HOI and N aOH
are employed, the final solution will contain only HOH, a small
quantity of undissociated N aCI molecules and N a+ and Cl- ions.
If one of the products of a reaction is removed the reaction
proceeds to completion readily; e.g., when KNO a in solutio:p. is
treated with HCI solution the equilibria represented by: equations
(1) and (2) exist prior to their admixture.
(1)
KNO a
K+
(2)
HCI
Cl-
+NOa
+H+
U(3)
U(4)
KCl
HNO a
When the two solutions are mixed, equilibria become estaplished between the four reactions represented. If the solution
is boiled, HNO s is volatilized as fast as it is formed in reaction (4),
removing H+ and NO; ions, thereby causing reactions (1) and
(2) to go to completion. Upon evaporation of all of the water,
KCl remains.
Under Precipitation Methods numerous examples will be
encountered of the use of an excess of one component of a reaction
and of the removal of one of the products in the form of an insoluble precipitate as methods of forcing reactions to completion.
Indicators.-The indicators commonly used to determine the
end point in neutralization processes are as a rule complex
organic compounds capable of existing in t~o forms of different
color which are mutually convertible, one into the other, at
given H+ and OH- ion concentrations. Three theories have
been proposed to explain the change in color of indicators which,
briefly stated, are as follows:
'
1. The physico-chemical theory attributes the color to certain
ions an increase of which causes the appearance of a new color,
and the decrease of which causes the disappearance of a color or
the appearance of a different color.
71
2, The organic theory attributes the color of indicators to

certain groupings of the elements in a compound, and the change
in color to. change in molecular structure.
?: The colloidal theory assumes that indicators form colloidal
solutions the change in color of which is dependent upon change in
size.of the colloidal particle.
The point at which an indicator changes color in any given
titration is dependent on the hydrogen ion concentration of the
solution and may not be indicative of the absolute neutrality
or completi.on of a reaction. The accompanying table of the
commonlY' used indicators gives the H+ ion concentrations
expresse_d in terms of pH between which color changes occur.
TABLE VII.-COMMONLY USED INDICATORS
Color
Indicator
Bromcresol purple ..............

Bromphenol blue ...............
Bromthymol blue ...............
Cochineal. .....................
Congo red .....................
Litmus ........................
Methyl orange .................
Methyl red ....................
Phenol red ...................
Phenolphthalein ................
Thymol blue ..................
pH range
5.2to6.8
3.0to4.6
6.0 to 7.6
4.8 to 6.2
3.0 to 5.0
4.5 to 8.3
3.1 to 4.4
4.2 to 6.3
6.8to8.4
8.0 to 9.8
8.0 to 9.6
Acid
Alkaline
Yellow
Yellow
Yellow
Yellow
Blue
Red
Pink
Red
Yellow
Colorless
Yellow
Purple
Bluish violet
Blue
Lilac pink
Reddish yellow
Blue
Yellow
Yellow
Red
Red
Blue
This table shows that methyl orange exhibits _its acid color,
pink, at a pH of 3.1 and its alkaline color, yellow, at a pH of 4.4,
while between these pH values the color undergoes transition
from one shade to the other. At a pH of 4.4 the solution is
slightly acid, since at neutrality pH = 7.
It would appear that an indicator which changes color exactly
at the neutral point, pH = 7, would be required in every case,
but this is not true. When a strong acid, such as hydrochloric
acid, is titrated with a strong base, like sodium hydroxide, the
change in concentration of hydrogen ion becomes very rapid as
72
the stoichiometric point is approached so that very small amounts

of acid or alkali in excess carry the concentration of hydrogen
ion far to one side of the neutral point, and the indicator will
give a sharp end point. If a solution of 25 cc. of 0.1 N HCl
is titrated with a solution of 0.1 N NaOH, the equivalent point
or stoichiometric point would be reached when 25 cc. of 0.1 N
NaOH had been added. Theoretically, the solution at the end
of titration should be identical in composition with a solution
prepared by dissolving a known weight of sodium chloride in
the calculated amount of solvent. When methyl orange is used
as indicator, the end point, as shown by change of color, appears
when about 24.95 cc. of 0.1 N NaOH have been added or while the
solution is still slightly acidic, but if phenolphthalein is used as
indicator, the end point appears when about 25.05 cc. of 0.1 N
NaOH have been added or when the solution is slightly alkaline.
The deviation of the end point from the stoichiometric point is
so slight that it is negligible in this case.
When a weak acid is titrated with a strong base, the change
in hydrogen ion concentration in the vicinity of the stoichiometric point is not so abrupt; e.g., if 25 cc. of: 0.1 N acetic acid are
titrated with 0.1 N NaOH, 25 cc. of the alkali should be required
to attain the stoichiometric point. When methyl orange is
employed as indicator, the end point is reached when about
17.5 cc. of 0.1 N NaOH have been added, but if phenolphthalein
il5 used the end point is reached when practically '25 cc. of 0.1 N
NaOH have been added. Obviously methyl orange would not
be a satisfactory indicator in the above case.
Acetic acid titrated with sodium hydroxide forms sodium
acetate: CHaCOOH
NaOH
CHaCOONa
HOH. The
acetate ions from the highly ionized sodium acetate force the
reaction to the left CHaCOOH
H+ + CHaCOO- before an
equivalent quantity of NaOH has been added, and the hydrogen ion concentration becomes very small before all of the hydrogen of acetic acid has been replaced. In this case methyl orange
which changes color at a relatively high hydrogen ion concentration 10-4.4 will show alkaline color gradually long before the
equivalent quantity of alkali has been added, and the end point
will not be sharp. Phenolphthalein, however, does not change
color until the true neutral point pH = 7 has been passed, and
73
a drop of alkali will than reduce the concentration below [H+] =

10-9 8 or pH = 9.~ where phenolphthalein exhibits a change of
color.
A weak bas~ titrated by a strong acid, e.g., ammonium hydroxide titrated with sulfuric or hydrochloric acid, exhibits the
reverse of the .above condition, as would be expected, and methyl
orange is a suitable indicator while phenolphthalein is not.
Rules for the Use of Indicators.-The following rules should
be observed ih the use of indicators:
1. Use two drops of indicator solution: for a titration unless
otherwise directed.
2. When 'It strong acid is titrated with a strong alkali or a strong
alkali with a strong acid, either methyl orange, methyl red, or
phenolphthalein may be used.
3. When a weak acid is titrated with a strong alkali, use phenolphthalein as indicator.
4. When a weak alkali is titrated with a strong acid, use methyl
orange' as indicator.
5. PI. weak alkali should never be titrated with a weak acid or
vice ver'sa, since no indicator will give a sharp end point.
6. 'Fhe appearance of a color is more easily observable than is
the disappearance. Always titrate where possible, therefore, to
the appearance of a color.
The criteria of a good indicator for analytical purposes are
that there shall be a sharp contrast between the two colors which
it e~hibits in acid and alkaline media and that the change in
color shall take place over a very small range of hydrogen ion
concentration. Unless otherwise stated, each indicator solution
should be so adjusted that when 0.15 cc. of the indicator solution
is'added to 25 cc. of distilled water, 0.25 cc. of 0.2 N acid or alkali,
respectively, will develop the characteristic color changes.
Some Commonly Used Indicator Solutions.
Bromcresol Purple. Di - bromo-o-cresol-sulfon -phthalein.-A
solution of 0.1 Gm. of the dry powder in 9.5 cc. of 0.050 N
NaOH diluted to 250 cc. is used as indicator solution. Two to
four drops of indicator per 100 cc. of solution should be used in
titration unless otherwise directed.
Bromphenol Blue. Tetra-bromo-phenol-sulfon-phthalein.-A
solution of 0.10 Gm. of dry powder in 100 cc. of 50 per cent
74
QUANTITATIVE PHARMACEUTICAL CH,EMISTRY
alcohol is used as indicator. Four to six drops <tf indicator per

100 cc. of solution should be used in titrations.
Bromthymol Blue. Di - bromo - thymol- sulfon iPhthalein.-A
solution of 0.10 Gm. of dry powder in 100 cc. of 50 ph cent alcohol
is employed as indicator. Two to four drops of indicator solution
per 100 cc. are used in titrations.
Cochineal.-Cochineal consists of the dried' female fi\sect
Coccus cacti. The indiclltor constituent is q,arminic 9,cid.
Cochineal indicator solution is prepared by macerliting 1 Gll}. of
the powdered, dry insects for three days with 25 per cent alcohol
and filtering. In titrations, 2 to 4 drops of indicl}tor solution
should be used for each 100 cc. of titration mixture l\nless otherwise directed.
This indicator is suitable for the titration of alkaliesJ.nd alkali
carbonates, ammonia, and many alkaloids and inorga:Q.ic acids,
but it should not be used for the titration of organic acids.
Congo Red. Diphenyl-disazo-bis-a-naphthyl-amine-4i>ulfonic
Acid.-The indicator solution is prepared by dissolving \50 mg.
of the dye in 100 cc. of 70 per cent alcohol. This indiC)ator is
used in the Kjeldahl ammonia determination. In titrations,
3 to 4 drops of indicator are used for each 100 cc. of solutton.
Litmus, or tournesol, consists of a blue pigmentlobtained,from
various species of Roccella De Candolle py fermentation. The
pigment mixed with chalk or other diluent is marketed in cu})es or
fragments. The blue coloring matter consists chiefly of azolitmin. The indicator solution is prepared by extracting 25 Gm. of
the solid commercial product with three successive portions of
100 cc. of boiling alcohol to remove red coloring matter. i'he
undissolved residue is extracted with cold distilled water to
remove soluble carbonates. These extractives are discarded.
The residue is then treated with 125 cc. of distilled water, boiletl,
cooled, and filtered. The filtrate constitutes the indicator
solution.
Other indicator solutions are superior to litmus for titration
purposes, but litmus test solution serves well for rough qualitative
tests of hydrogen ion concentration.
Methyl Orange. The Sodium Salt of Dimethylamino-azo-benzene Sulfonic Acid.-The indicator solution is prepared by
dissolving 0.10 Gm. of the dry salt in 100 cc. of distilled water.
PRINfIPLES OF VOLUMETRIC ANALYSIS
75
Two to four drops per 100 cc. of reaction solution are employed
in titrations.
The color ({hange is very sharp if too much indicator is not
used. Meth:lll orange is used frequently in the titration of strong
acids and strqng alkalies and especially in the titration of weak
bases, e.g., NH 40H. It is also a good indicator to use in the
titration of th'e salts of weak acids such as carbonates, borates,
sulfides, etc., with a strong acid, since in these cases the acids
liberated in t~e titration reaction are too little ionized to affect
the indicatorj It does not give the proper end point in the titration of alkal6ids or organic acids. This indicator should never
be employe~ in the titration of alcoholic solutions, hot solutions,
or very dilnte solutions.
Methyl Red. o-Carboxy-benzene-azo-dimethyl-aniline.-The
indicator(solution is prepared by dissolving 0.10 Om. of the dry
dye in 1.00 cc. of 95 per cent alcohol. Two to four drops of the
resulting solution are used per 100 cc. in titrations. Methyl
red in{licator is especially useful in titrating ammonia, weak
bases, ,and alkaloids, but it is not suitable for the titration of
weak organic acids.
Phenol Red. Phenol-sulfon-phthaJein.-The indicator solution ,Is prepared by triturating 100 mg. of the dry powder in a
mor-ar with 14.5 cc. of 0.020 N NaOH until solution is complete
and <liluting the resulting solution to 250 cc. with recently boiled
distilled water. After the removal of any precipitate formed
upOll dilution, the solution is ready for use. In titrations, 3 to
6 drops of this indicator should be used for each 100 cc. of
sollJtion.
lfhenolphthalein indicator solution is prepared by dissolving
1 Gm. of phenolphthalein in 100 cc. of 95 per cent alcohol. Two
to three drops of this indicator should be used per'100 cc. of solution in titrations unless otherwise directed.
Phenolphthalein is an excellent indicator to use in the titration of weak organic and inorganic acids, most alkalies, and alkali.
salts, but it is not satisfactory for use in the titration of ammonia, alkaloids, or cold solutions of carbonates and bicarbonates.
It fimctions well in alcoholic solutions.
Thymol Blue. Thymol-sulfon-phthalein.-The indicator solution is 'prepated by dissolving 40 mg. of the dry substance in
76
100 cc. of alcohol. Two to four drops of indic~tor should be

employed in titrations per 100 cc. of reaction solution.
Sensitivity of Indicator Solutions.-Indicator solutions should
be tested for sensitiveness. This is done by addipg 0.15 cc. of
indicator solution to 25 cc. of distilled water and titrating the
resulting solution with 0.02 N alkali or acid. Upon the addition
of 0.25 cc. of the standard alkali or acid solution,lthe respective
characteristic color should develop.
1. Define the following terms: (a) stoichiometric point, (b; end point in
titration, (c) indicator, (d) buffer.
2. If a solution has a hydrogen ion concentration of 10- 9 \ what is its
hydroxyl ion concentration?
3. In a solution having a hydrogen ion concentration of [H+] = 10- 6 . 6 ,
what would be the pH value? Would the solution be acid or al~line?
4. What indicator is best suited to the following titrations? Why?
(a) alkali carbonate with sulfuric acid, (b) acetic acid with sodium hydroxide,
(c) ammonium hydroxide with hydrochloric acid, (d) organic acids with
sodium hydroxide.
5. How may the sensitiveness of an indicator be determined?
6. Why is the methyl orange a good indicator to use in the titrlttion of
NH.OH?
Standard Solutions.-The concentration of stalldard sohlt.ions

can be expressed in various ways as follows:
A normal solution is one which contains 1 Gm. equivalent
weight of reagent dissolved in sufficient solvent to make 1,000 cc.
of solution.
Molar solutions are those which contain in 1,000 cc. a gfammolecule equivalent of the reagent, irrespective of the valency
of the molecule. Thus, a molar solution of sulfuric acid would
contain 98.08 Gm. of H 2S0 4 in a liter of the solution, and a molar
solution of potassium dichromate would contain 294.21 Gm. of
K2Cr207 in a liter of the solution. Solutions containing in
1,000 ee. one-tenth of a gram-molecule of the reagent are designated "tenth-molar," MilO, and other molarities are similarly
indicated.
A molal solution is one which contains 1 mole or Gm.-moleculll.r
weight of reagent dissolved in 1,000 Gm. of solvent.
The concentration of official standard solutions, volumetric
solutions, is expressed in terms of normality or molarity. It is
77
very important that what is meant by a normal and a molar

solution be clearly; understood. A normal solution contains one
gram equivalent weight of reagent per liter of solution, i.e., that
quantity of reagent which is chemically equivalent to 8.000 Gm.
of oxygen or 1.0078 Gm. of hydrogen per 1,000 cc. The gram
equivalent weight, in neutralization reactions, is equal to the
gram-molecular weight (molecular weight of substance in grams)
divided by the number of H+ or OH- ions entering into or formed
in the reaction. From the above definition of a normal solution,
it is obvious that a normal acid solution contains 1.0078 Gm. of
available hydrogen ion per liter; thus, a normal solution of
hydrochloric acid contains 1 mole, 36.47 Gm. of HCI, and a
normal solution of sulfuric acid contains 0.5 mole, 49.04 Gm. of
H 2S0 4, per 1,000 cc. of solution, becaUJ3e those weights of the
respective acids contain 1.0078 Gm. of available hydrogen. A
normal alkali solution contains sufficient available hydroxyl
ion in 1,000 cc. to combine with 1.0078 Gm. of hydrogen ion or
17.0078 Gm. e.g., 1 mole, 40.00 Gm. of sodium hydroxide or
0.5 mole, 157.75 Gm. of barium hydroxide per 1,000 cc. of solution. A normal oxidizing solution has an oxidizing value of
8.000 Gm. of oxygen equivalent to 1.0078 Gm. of hydrogen
per 1,000 cc.; i.e., 2KMn04 used as an oxidizing agent gives up
5 atoms of available oxygen equivalent to 10 atoms of available
hydrogen, therefore a normal solution will contain 0.2 mole,
31.606 Gm. of KMn04, per 1,000 cc. of solution. It is evident
that equal volumes of normal alkali and acid solution when mixed
will exactly neutralize one another if they react quantitatively
and that equal volumes of normal oxidizing and reducing solutions will exactly use up one another.
The letter N is used to indicate normality. The normality of
a solution may be represented in two ways; i.e., a solution of
exactly normal strength is indicated by 1 N or liN (normality =
1.0000), a double normal ~trength by 2 N or 21N (normality = 2.0000), a half or seminormal by 0.5 N or N 12 (normality = 0.5000), a decinormal by 0.1 or N /10 (normality =
0.1000), and a centinormal by 0.01 or N 1100 (normality =
0.0100), etc.
In practical analytical work, it is unnecessary to prepare or
maintain solutions exactly normal or tenth-normal, etc. It is
78
usually easier and just as satisfactory to determine the normality

of a solution which closely approaches the value of an exactly
normal solution. The relationship of an approximately normal
solution to an exactly normal solution is expressed by a factor,
often called the normality factor; e.g., a normal solution of sodium
hydroxide contains 40.00 Gm. (the gram equivalent weight)
per 1,000 cc., and an approximately normal solution contains
40.42 Gm. per 1,000 cc. The latter is then 40.42/40.00 = 1.0105
normal and any number of cubic centimeters of this solution used
can be expressed in terms of exact normality by multiplying by
this factor; thus 20.1 cc. represents 20.1 X 1.0105 = 20.31 cc. of
1 N solution.... The standard alkali solution should be labeled
1.0105 N NaOH. The normality factor expresses the ratio
between the weight of reagent present in a liter of solution and
the weight that would be present if the solution were exactly
normal. . Usually the factors of volumetric solutions are limited
to four significant figures.
The standard acid solutions used in acidimetry and alkalimetry are usually prepared from hydrochloric or sulfuric
acid. Either acid may be used in most titrations, but hydrochloric is preferable to sulfuric acid in the titrat10n of compounds which yield a precipitate with the latter, such as barium
hydroxide, and sulfuric acid is preferable in hot titrations,
since there would be danger of loss of hydrochloric ,acid due to
volatilization.
The standard alkali solutions commonly used are sodium
hydroxide, potassium hydroxide, and barium hydroxide. These
hydroxide solutions absorb carbon dioxide from the air, thereby
changing rapidly in concentration. Sodium and potassium
hydroxides may become contaminated with carbonates which
impair their usefulness due to the liberation of carbon dioxide
during acid-alkali titration. Barium hydroxide solutions remain
free from carbonates, since absorbed carbon dioxide is precipitated as insoluble barium carbonate; this, however, decreases
the concentration of barium hydroxide in the solution. Alkali
solutions should be prepared carbonate-free, and they should be
protected from carbon dioxide by means of a soda-lime absorption tube. All alkali solutions should be restandardized frequently.
79
Exercise 10
Preparation and Standardization of Hydrochloric Acid Solution.-Normal hydrochloric acid solution may be standardized
against pure sodium carbonate, standard solution of sodium
hydroxide, or gravimetrically by precipitating and weighing the
chloride ion as silver chloride.
Object.-To Prepare and Standardize Normal Hydrochloric
Acid.
Materials Required.-Hydrochloric acid.
4 Gm. of anhydrous sodium carbonate.
Procedure.-l. Dilute 95 cc. of hydrochloric acid with sufficient distilled
water to make 1,000 cc., and mix thoroug}lly.
Since"hydrochloric acid, specific gravity 1.175, contains about

36 per cent of HCI, the resulting solution should be slightly
stronger than normal. Thus, 95 X 1.175 X 0.36 = 40.18 Gm.
HCl. Since this quantity of HCI is dissolved in 1,000 cc., the
solution sh~)Uld contain approximately, 40 Gm. HCl.
2. Weigh about 4 Gm. of pure sodium carbonate on a rough balance and
place the salt in a porcelain dish of suitable capacity. 'Heat the carbonate
for 1 hr. over a low Bunsen flame or in an oven at 270C. and allow it to cool
in a. desiccator. Weigh accurately two portions of about 1.5 Gm. each of the
cool dry salt, recording the weights.
Although pure sodium carbonate can be purchased, the salt

contains varying amounts of water of crystallization and moisture. When it is dried, the absorbed moisture is driven off
as well as any water of, crystallization, since sodium carbonate
becomes anhydrous when heated to 100C.
3. Transfer each of the samples to 250 cc. Erlenmeyer flasks and proceed
to titrate them separately as follows: Add about 100 cc. of distilled water
~nd 2 drops of methyl orange T.S. and shake until the carbonate is completely dissolved. Fill the glass-stcppered burette exactly to the mark
with the acid solution and run acid 'slowly into the flask with continuous
shaking until the solution just becomes pink in color. A white porcelain
plate or a piece of white paper placed beneath the flask renders the end
point more easily discernible. Wash down the inside walls of the flask
with distilled water from a wash bottle, and if the solution becomes yellow,
add acid drop by drop until the faint pink color is restored, taking care that
the last drop which adhered to the burette tip is introduced into the solution.
80
When titration is complete, read the burette accurately and record the
volume of acid used in the titration.
Assuming that a sample of sodium carbonate weighing 1.6250

Gm. required 30.20 cc. of acid in the titration, the calculations
are made as follows:
If the acid were exactly normal, according to the definition of
d 1 mole 2Na 2CO a =
.
a normaI soIutlOn,
1,000 cc. woul neutralIze
10~99
53 Gm. sodium carbonate, and the sample used would
require:
53 : 1,000: : 1. 6250 : x
1625.0
x = ---s3 = 30.66 cc.
Since only 30.20 cc. of acid were required in the titration, it is
evident that the acid solution is stronger than normal, and the
factor is 30.66/30.20 = 1.0152 N. The bottle containing the
acid should be labeled 1.0152 N HCl.
The calculation may also be made by the method of equivalents, namely,
Na 2 CO a + 2HCI = 2NaCI + H 2 0 + <t0 2
2) 105.99
2)72.94
36.47 = 1,000.cc. N HCI
53 Gm.
0.053 Gm. Na 2CO a = 1 cc. of N HCl, and 1 Gm. of Na 2CO a is
equivalent to 1,000/53 = 18.87 cc. of N HCl. Therefore,
1.6250 Gm. will require 1.6250 X 18.87 = 30.66 cc. of N HCI, and
30.66/30.20 = 1.0152 N, the factor of the acid. The solution
may be used for all practical work as standardized, the number
of cubic centimete!s used being multiplied by the factor to reduce
the solution to terms of exact normality.
If an exactly normal solution is wanted, the dilution required
may be calculated as follows:
30.66 - 30.20 = 0.46 cc. of water to be added to each 30 2 CC.
of the standard solution to make it exactly normal. To make
1,000 cc. of normal solution, it would be necessary to dilute with
water in the ratio, 0.46: 30.66: :x: 1,000 = 15.00 cc. of water in
1,000 cc. of the diluted solution. Measure 15.00 cc. of distilled
81
water into a dry 1,000 cc. flask, fill to the mark with 1.0152 N
Hel, mix the solution well, and verify the normality of the solution by titratiQn agamst weighed samples of sodium carbonate.
Volumetric solutions of hydrochloric acid may be standardized
gravimetrically by determining the amount of chloride ion
present as silver chloride, as described under gravimetric determination of ch~orides, using 10 cc. portions of the acid as samples.
Ten cubic centimeters of N Hel is equivalent to 1.4334 Gm. of
Agel. If a 10 cc. portion of acid is found to yield 1.4634 Gm. of
Agel, the acid is 1.4634/1.4334 = 1.0210 normal.
Solutions of hydrochloric acid may also be standardized by
titration against standard solutions of KOH or NaOH. Thus _ii
25.20 cc. of 0.9505 N NaOH is required to exactly neutralize
25.00 ce. 9f acid, the factor of the acid is 25.202~0~9505 = 0.9531
normal.
The normal solution is too strong for many assay procedures,
so 0.5 N, 0.2 N, 0.1 N, etc., solutions are employed. These may
be prepared by dilution of the normal solution in the proper _
ratio, but the resulting solution should always be restandardized.
Standard solutions of hydrochloric acid preserved in tightly
stoppered, alkali-free, glass bottles retain their strength indefinitely.
Exercise 11
Preparation and Standardization of Alkali Solutions.-Solutions of sodium hydroxide or potassium hydroxide may be
standardized against standard solution of hydrochloric or sulfuric acid or by titration against an accurately weighed quantity
of pure potassium bitartrate or potassium biphthalate.
Object.-To Prepare and Standardize Normal Sodium Hydroxide Solution.
Materials Required.-About 50 Gm. of sodium hydroxide.
A saturated solution of barium hydroxide.
A standardized solution of hydrochloric acid.
Procedure.-l. Weigh about 50 Gm. of sodium hydroxide on a rough
balance and dissolve it in about 1,000 cc. of distilled water. Add slowly
with stirring a sat~rated solution of pure barium hydroxide until no further
precipitation occurs (2 to 3 cc.). Allow the precipitate to subside for about
10 hr. and decant the liquid through a filter into a hard glass bottle or a
82
bottle the inside of which has previously been coated with paraffin. Close
the bottle tightly with a rubber stopper provided with a soda-lime tube.
More than a mole, 40.00 Gm., of sodium hydroxide is weighed

out, since it is hygroscopic, and if only a mole were used the resulting solution would be below normal strength. The solution is
treated with barium hydroxide to precipitate any carbonate
formed through the action of carbon dioxide on the sodium
hydroxide. The solution is preserved in a tightly stoppered
bottle fitted with a soda-lime tube to protect it from the carbon
dioxide of the air. Solutions which contain carbonate are not
suitable for titration with phenolphthalein as indicator, but when
methyl orange is used the results are the same as if all the sodium
were present combined as hydroxide.
2. Accurately measure 30 cc. of normal hydrochloric or normal sulfuric
acid, dilute with 50 cc. of carbon dioxide-free distilled water, add 2 drops of
phenolphthalein T.S., and titrate with the sodium hydroxide solution to the
production of a permanent pink color. If the end point is passed in the
titration, add a few cubic centimeters more of acid and again titrate to
the end point. Titrate several portions of the alkali and from the mean of
the results calculate the normality of the sodium hydroxide solution.
A large volume, about 30 cc., of solution-is used for each

titration to minimize error from measurements. Every precaution should be taken to insure the correct ineasurerhent of the
volume of solutions. The burettes should be cleaned, freed from
air bubbles before titration, and read with the great~st possible
accuracy.
The solution can be made exactly normal by dilution in the
same manner as that explained under the preparation of normal
hydrochloric acid. Assuming that 30.00 cc. of alkali solution
required 30.15 cc. of 1.0119 N HCI, the factor of the sodium
hydroxide solution would be
30.15 ;01.0119 = 1.0170.
Standard solutions of potassium hydroxide can be prepared
in the same way using 1 mole, 56.10 Gm., of KOH per liter. Halfnormal and tenth-normal, etc., solutions can be prepared directly
or by proportionate dilution of stronger solutions, but they
should always be restandardized when prepared by the latter
method.
83
Sodium hydroxide or potassium hydroxide solutions can also be

standardized against Ei,tandard sulfuric acid or against accurately weighed samples of pure dry potassium biphthalate or
potassium bitartrate. The alkali converts the potassium acid
tartrate to potassium and sodium tartrate or potassium tartrate
when sodium or potassium hydroxide solution, respectively,
is used
KHC 4H 4 0 6
188.14
+ NaOH-KNaC H
4
06
+H
40.00
One thousand cubic centimeters of N NaOH contains 40.00 Gm.

of alkali and is equivalent to 188.14 Gm. of potassium bitartrate.
Therefore, 1 cc. of N NaOH is equivalent to 0.1881 qm. of
potassium bitartrate. From the above equivalent the normality
may be calculated.
Exercise 12
Object.-To Prepare and Standardize 0.1 N Solution of

Barium Hydroxide.
Materials Required.-18 Gm. barium hydroxide.
0.1 N hydrochloric acid.
Procedure.-Dissolve about 18 Gm. of pure barium hydroxide in 1,000
cc. of recently boiled, cooled, distilled water and filter the solution quickly
if turbid. Standardize the solution against N Hel using phenolphthalein
as indicator. It is not advisable to adjust the solution to exactly 0.1 N,
since it usually becomes turbid on dilution and deteriorates rapidly. The
solution should always be standardized before using. It should be protected
from carbon dioxide of the air in a well stoppered bottle fitted with a sodalime tube.
Barium hydroxide solutions change in concentration rapidly

because the absorption of CO 2from the air results in the formation
of insoluble barium carbonate as follows:
Ba(OH)2
+ CO 2-BaC0 + H 20
3
The explanation of the procedu~e and the calculations is similar

to that given under the preparation of normal sodium hydroxide
solution in the preceding exercise.
Exercise 13
Object.-To Prepare and Standardize Normal Sulfuric Acid.

Materials Required.-30 cc. sulfuric acid.
84
Normal sodium hydroxide solution.

Procedure.-Add 30 cc. of sulfuric acid in small portions to 1,020 cc. of
distilled water with constant stirring. Allow the solution to acquire room
temperature. Standardize the solution by titration against normal sodium
hydroxide solution using phenolphthalein as indicator. Calculate the
normality factor and write it on the label or adjust the solution to exact
normality.
Since sulfuric acid is a dibasic acid, 1,000 cc. of a normal

solution should contain 98.08/2 = 49.04 Gm. Sulfuric acid
(95 per cent) has a specific gravity of about 1.83 at 25C. Consequently, 30 cc. of the concentrated acid should contain about
30 X 1.83 X 0.95 = 52.15 Gm. H 2S0 4 The method of calculating the normality of the solution is similar to that described
under the preparation of normal hydrochloric acid (page 80).
The standardization of sulfuric acid solution can also be
effected by titration against pure sodium carbonate as described
under the standardization of hydrochloric acid. It may also be
standardized gravimetrically by precipitation and weighing the
sulfate ion as barium sulfate as described under the gravimetric
determination of sulfate in a soluble sulfate.
Standard solutions of sulfuric acid preserved in tightly stoppered, alkali-free bottles do not deteriorate.
1. Define the following terms: (a) normal solution, (b) molar solution,
(0) molal solution, (d) gram equivalent weight, (e) normality factor.
2. How much of each of the following reagents would be required to
make 500 CC. of 1 N, 0.5 N, 0.1 N, and 0.020 N solutions, respectively?
(a) HCI, (b) H 2S0 4, (0) NaOH, (d) Na,C03, (e) Ba(OH)..
3. Calculate the normality of each of the following: (a) a solution of
HCl containing 54.7 Gm. per liter, (b) a solution of Na,C0 3 containing 32
Gm. per 500 cc., (0) a solution of H 2SO. containing 9.8 Gm. per 200 cc., (d)
a solution of Ba(OH)2 containing 1.6 Gm. per 40 cc.
4. If 1.1200 Gm. of pure Na,C0 3 is neutralized by 40.2 cc. of HCI solution, what is the normality of the HCI?
6. If 20.5 cc. of alkali solution neutralizes 4.5020 Gm. of pure KH9.H.O.,
what is the normality of the alkali solution.
6. How much 0.1 N volumetric solution could be prepared from each of
the following? (a) 100 cc. of 0.6500 N'HCI, (b) 250 cc. of 0.1235 N H 2SO.,
(0) 50 cc. of 0.9850 N NaOH.
7. If 40 cc. of HCI solution yielded 0.2140 Gm. of AgCI upoh precipitation of the chloride ion as silver chloride, what was the normality of the HCI
solution?
85
8. 25 cc. of HOI solution is found to be equivalent to 0.8 Gm. of pure

sodium carbonate. How much of each of the following PUre reagents would
be exactly equivalent to 25 cc. of the hydrochloric acid? (a) NaHOO.,
(b) KOH, (c) Ba(OH)., (d) Oa(OH)2.
9. An unknown solution of alkali weighing 10.5420 Gm. required 36.40
co. of 0.0860 N acid for neutralization. Calculate the percentage of alkali
present if the unknown were Na200., Ba(OH)2, or NaOII, respectively.
10. How much 0.1240 N H 2S0 4 would be required to neutralize 25 cc. of
0.0500 N NaOH; 40 cc. of 0.1020 N NaOH; and 50 cc. of 0.5342 N KOH?
11. How many cubic centimeters of each of the following reagents are
required to make 500 cc. of 1.0000 N, 0.1000 N, 0.0200 N, 0.0100 N solutions,
respectively: (a) 36 per cent HCI, specific gravity 1.12, (b) 10 per cent HCI,
specific gravity 1.05, (c) 75 per cent H 2S0 4, specific gravity 1.67, and (d)
10 per cent H 2S0 4, specific gravity 1.07?
CHAPTER V
ALKALIMETRY
Alkali hydroxides and carbonates, etc., are usually titrated
directly with standard solutions of hydrochloric or sulfuric
acid using methyl orange as indicator. Phenolphthalein and
methyl red when used as indicators in such titrations are affected
by the acidic carbon dioxide liberated during titration so that the
end point appears before neutralization is complete. If phenolphthalein is used as indicator, carbonates must be removed by
precipitation before titration, or the titration mixture must be
boiled to expel the carbon dioxide formed during titration.
Direct titration is conducted by adding a standard reagent
solution in measured quantity to a substance in solution until the
end point, as shown by a change of indicator color, is reached.
Substances which are insoluble in water or which do not yield
a sharp end point that coincides with the stoichiometric point
upon direct titration are titrated residually.
Residual titration is conducted by treating the substance under
analysis with an amount of standard solution knorvn to be in
excess of that actually required to react with it completely;
the amount of standard solution in excess is then determined by
titration with another standard solution.
Exercise 14
Object.-Assay of Sodium Bicarbonate.

Materials Required.-About 8 Gm. of sodium bicarbonate.
Normal sulfuric acid.
Procedure.-" Dry about 3 Gm. of Sodium Bicarbonate to constant_weight
over sulfuric acid, weigh accurately, mix it with 25 cc. of distilled water, and
titrate with normal sulfuric acid, using methyl orange T.S. as the indicator.
Each cubic centimeter of normal sulfuric acid is equivalent to 0.08400 Gm.
of NaHCO a."
It is a general rule that when the substance to be assayed is a

solid, the accurately weighed quantity to be used should be dis86
ALKALIMETRY
87
solved in sufficient water to make the solution of about the same

concentration as that of the acid to be used in the titration.
Methyl orange is us~d as indicator because phenolphthalein and
most other indicators are affected by the carbonic acid liberated
in the reaction showing a change of color before the reaction is
complete:
2NaHCO a
2(84.00)
+ H2S04~Na2S04 + 2H20 + 2C0 2

98.08
Since sulfuric acid has a hydrogen equivalent of two, 1 cc.

of its normal solution is equivalent to 2 :~~ooo = 0.0840 Gm.
NaHCO a
Calculate the percentage purity of the sample assayed.
1. Write ionically all equations for all reactions that occur in the above
assay.
2. Could normal acetic acid be used to titrate the bicarbonate in place of
sulfuric acid?
3. Look up the pH at which methyl orange exhibits its acid color and
explain. why it is a good indicator to use in this titration.
4. If exactly 3.0 Gm. of NaHCO s are dissolved in 25 cc. of distilled water
in the above assay, what would be the normality of the solution? Does
the normality conform to the general rule stated under Explanation of
Procedure?
6. Enumerate the official carbonates and bicarbonates assayed by similar
methods.
Exercise 16
Object.-Assay of Sodium Hydroxide.

Materials Required.-1.5 Gm. sodium hydroxide.
Procedure.-l. "Dissolve about 1.5 Gm. of Sodium Hydroxide, accurately weighed, in about 40 cc. of recently boiled and cooled distilled water.
Cool the solution to 15C. and 'titrate with normal sulfuric acid, using
phenolphthalein T.S. as the indicator. At the discharge of the pink color
of the indicator, record the volume of acid solution required."
The base, NaOH, behaves in the same manner when either

phenolphthalein or methyl orange is used as the indicator.
Consequently, the end point obtained with phenolphthalein as
88
the indicator represents complete neutralization of all of the

NaOH as represented by the equation:
2NaOH
2(40.00)
+ H2S0r--~Na2S04 + 2H 20
98.08
142.05
In a cold solution, with phenolphthalein indicator, the end

point of titration of sodium carbonate with standard H 2S0 4 is
exhibited when the solution is one-half neutralized or when the
Na2CO S is transformed into NaHC0 3
2Na 2CO S
2(105.99)
+ H 2S0 4---Na 2S04 + 2NaHCO s

2(84.00)
2. "Add 3 drops of methyl orange T.S., and continue the titration until
the production of a permanent pink color. Each cubic centimeter of total
sulfuric acid consumed is equivalent to 0.0400 Gm. of NaOH. Each cubic
centimeter difference between the number of cubic centimeters of normal
sulfuric acid consumed in the methyl orange and phenolphthalein titrations
is equivalent to 0.1060 Gm. of Na 2CO a."
The base NaHCO a is practically neutral toward phenolphthalein because the hydrogen ion concentration of the weakly
ionized HCOa ion (HCO a H+
C03') is about the same
(1 X 10- 9) as that necessary to change the color of this indicator.
A minute amount of H 2S0 4 is sufficient, therefore! to indicate a
neutral solution by the discharge of the pink color. When methyl
orange is used as indicator, it does not change color from alkaline
to acid until a hydrogen ion concentration of about 1 X 10- 4
is obtained. This will occur only when all of the bicarbonate
has been neutralized. The reaction is indicated by the equation:
2NaHCO s + H 2S0 4---Na 2S04

2(84.00)
+ 2C0 + 3H 0
2
The method of calculation is as follows: If a 1.0000 Gm.

sample of NaOH r.equired 20.80 cc. of N H 2S0 4 to titrate to the
end point with phenolphthalein indicator and an additional
0.95 cc. of N H 2S0 4 to titrate to the end point with methyl
orange indicator, the total alkali calculated as NaOH would be
20.80 + ~. ~go~ 0.0400 X 100 = 87 per cent.
The volume of
N H 2S04 required to titrate the NaHCO s, after the titration to

an end point with phenolphthalein, represents one-half of the
acid that would be necessary to neutralize the Na 2CO a originally
ALKALIMETRY
89
present. The equivalent weight of Na 2 CO a is 105.99/2 X

1000 = 0.053 Gm. per cubic centimeter of N H 2S0 4 Consequently, by doubli'ng the equivalent, the result gives the per cent
0.95 X 0.106
1.0000
X 100 = 1.01 per cent. The
of Na 2 CO a, e.g.,
same result would be obtained by doubling the volume of acid
used and retaining the equivalent 0.053. (See calculation of
K 2CO a in solution of potassium hydroxide, N.F. VI.)
1. What volume of 0.5 N H 2SO. will be required to titrate 1.1124 Gm.
of pure N a 200. using phenolphthalein as indicator in a cold solution? What
volume will be required to titrate the same weight of sample using methyl
orange as indicator?
.
2. A sample weighing 1.0202 Gm. and consisting of a mixture of equal
parts of,Na 200, and NaHOO, is titrated with 0.8242 N H 2S0 4, with phenolphthalein in cold solution, and then with methyl orange. What volumes of
the H ,SO. solution will be necessary in each titration?
3. A 25.00 cc. sample of KOH solution required 22.25 cc. of N H 2SO. to
titrate to the end point with phenolphthalein indicator in the cold and 4.45
cc. of the same acid to titrate to the end point with methyl orange indicator.
Calculate the per cent of total alkali as KOH and the per cent of K 200, in
the solution.
4. Oalculate the per cent of Na,OO, and of NaHOO. in a 1.2500 Gm.
sample which on titration with 0.2000 N HOI required 36.25 cc. with phenolphthalein indicator and an additional 42.80 cc. with methyl orange indicator.
Exercise 16
Object.-Assay of Sodium Salicylate.

Materials Required.-2 Gm. of sodium salicylate.
100 cc. of ether.
Bromphenol blue, T.S.
Procedure.-1. "Transfer about 2 Gm. of Sodium Salicylate, previously
dried to constant weight at 1000. and accurately weighed, to a tall beaker
of about 300-cc. capacity, and add 75 cc. of ether and 10 drops of bromphenol blue T.S. Titrate the mixture with half-normal hydrochloric acid,
mixing intimately the aqueous and ethereal layers by vigorous stirring until
a permanent, pale green color is produced in the aqueous layer."
The sodium salicylate reacts with the added acid to form free
salicylic acid and sodium chloride as follows:
C 6H 4.OH.COONa
160.04
+ HCl~C6H4.0H.COOH + NaCl
138.05
gO
The salicylic acid dissolves in the ether layer almost as rapidly

as it is formed when the ethereal and aqueous layers are well
mixed.
2. "Transfer the contents of the beaker to a small separator, and draw off
the aqueous layer into a small flask. Wash the ethereal layer once with 5 CC.
of distilled water, and add this to the aqueous layer. Add 20 cc. of ether to
the combined aqueous solutions, and mix intimately. Continue the titration
with vigorous shaking until a permnnent, pnle green color is produced in the
aqueous layer. Each cubic centimeter of half-normal hydrochloric acid is
equivalent to 0.08002 Gm. of C eH 4.OH.COONa."
The separated ethereal layer is washed with water to remove

small amounts of sodium salicylate which may remain dissolved
therein. Upon washing the combined aqueous solutions and
mixing intimately, any remaining salicylic acid partitions itself
into the ethereal layer leaving only the Hel in the aqueous
layer to influence the end point.
1. Why is bromphenol blue a good indicator to use in the above assay?
2. Compare the above method of assay for sodium salicylate with that for
the assay of tablets of sodium salicylate. Which method is more simple and
rapid? What component of the sodium salicylate molecule\ is assayed for
in each case? On the basis of the therapeutically active component of the
molecule, which method is preferable?
3. List the official salicylate and benzoate compounds wit!). the indicator
used for each. Explain why the indicators used are or are not suitable.
4. Write equations for the reactions that take place in the assay of:
(a) sodium benzoate, (b) elixir of sodium salicylate, (c) strontium salicylate.
5. Show how the equivalent is derived for each of the assays in question 4.
RESIDUAL TITRATION METHODS
Residual titration or "back titration" is frequently used when

a reaction proceeds slowly or when the substance to be assayed
does not give a distinct, sharp end point with an indicator by
direct titration. Residual titration is carried out by diss'olving
the substance under examination in an accurately measured
quantity of standard solution known to be in excess and titrating
the excess of the latter with another standard solution. Numerous examples of this method of titration will be met with in
subsequent exercises.
91
ALKALIMETRY
II
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92

Exercise 17
Object.-Assay of Zinc Oxide.

Materials Required.-1.5 of zinc oxide.
Normal sodium hydroxide.
Procedure.-" Digest about 1.5 Gm. of freshly ignited Zinc Oxide, accurately weighed, _with 50 cc. of normal sulfuric acid until solution is complete. Then titrate the excess of sulfuric acid with normal sodium hydroxide,
using methyl red'-T.S. as indicator. Each cubic centimeter of normal
sulfuric acid corresponds to 0.04069 Gm. of ZnO."
The zinc oxide is digested with N H 2S0 4 to effect complete

solution. The excess acid is then determined by titration with
NNaOH:
~\~~oo
ZnO
H2S04~ZnS04
H 20
81.38
H 2S04 + 2NaOH~Na2S04 + 2H 20
=
0.04069 Gm. ZnO equivalent to 1 cc. N H 2S0 4
Calculate the percentage of zinc and the percentage of zinc oxide

in the sample assayed.
1. Explain why zinc oxide is soluble in sulfuric acid, employing the
solubility product principle.
2. Calculate the amount of ZnCO a, ZnS04, and ZnCh equivalent to 25 cc.
of 0.0245 N H.S0 4.
3. How much 1.0520 N H 2S0 4 would be required to react quantitatively
with 1.3560 Gm. of ZnO if the ZnO were 90 per cent pure?
Exercise 18
Determination of the Purity of the Alkali Salt of an Organic

Acid. The assay of the acetates, citrates, tartrates, etc., of alkali
metals is based upon their conversion by ignition into the corresponding carbonates. All of the alkali combined as carbonate is
determined by titration in the usual way and from the amount
of carbonate found, the quantity of organic acid salt originally
present may be calculated. The salt should, as a rule, be dried
previous to ignition in order that calculations may be made on a
moisture-free basis.
Object.-Assay of Potassium and Sodium Tartrate.
93
ALKALIMETRY
Materials Required.-2 Gm. of potassium and sodium tartrate.
0.5 N sulfuric acid.

Procedure.-l. "Heat about 2 Gm. of the salt, accurately weighed, in a
platinum or porcelain crucible, heating at first very gently, t:qen gradually
raising the temperature until the salt is thoroughly carbonized. (CautionDo not use platinum crucibles for lithium salts.) The final temperature
must not exceed a dull red heat and the flame of the burner must not come
in contact with the carbonized mass."
The salt is heated slowly at first, since the sample swells and
fuses with concomitant decomposition. If strong heat is applied
in the initial stages of ignition, there may be loss of a portion of
the sample through decrepitation or spattering. After the mass
in the crucible is partially charred and white fumes are no longer
evolved, it is ignited to dull redness. If heated too strongly or
if the flame comes in contact with the carbonized mass, the alkali
carbonate may be converted into the oxide.
2. "After allowing the carbonized mass to cool, disintegrate it with the
aid of a stout glass rod and transfer thc mass and crucible to a beaker. Add
50 cc. of distilled water and 50 cc. of half-normal sulfuric acid, cover the
beaker with a watch glass and boil the contents for thirty minutes. Then
filter the solution and wash the residue with hot distilled water until the
washings cease to redden blue litmus paper. Now determine the residual
acid in the cooled filtrate by titration with half-normal sodium hydroxide,
using methyl orange T.S. as the indicator. The volume of half-normal
sulfuric acid consumed, multiplied by the proper equivalent of the salt,
represents the quantity of the salt present in the quantity taken."
The fused residue is broken up by' means of a strong glass rod

over glazed, black paper and transferred with the crucible into
a 250 cc. beaker. About 50 cc. of distilled water and 50 cc. of
sulfuric acid are added along the glass rod, and the mass is completely dissolved.
The reaction taking place upon ignition with free advent of
oxygen from the air may be represented as follows:
KNaC4H406~KNaCOa
210.12
+ 3C0 2 + 2H20
\.
and upon the addition of an excess of acid the double carbonate

is converted into double sulfate:
KNaCO a + H2S04~KNaS04
122.1
98.08
+ CO 2 + H 20
94
The volume of acid in excess of that necessary to react with the

carbonate residue is determined by titration with standard alkali
solution. Thus, if 12.20 cc. of 0.5 N NaOH is required to neutralize the excess acid, 37.80 ee. of the 0.5 N H 2S0 4 must have reacted
with the carbonate in the residue.
One mole of the tartrate upon ignition forms one mole of the
carbonate which is equivalent to one mole of H 2S0 4 Sulfuric
acid has a hydrogen equivalent of two, and the standard solution
used is 0.5 N. Each cubic centimeter of 0.5 N acid consumed in
the titration is, therefore, equivalent to:
210.12
2 X 2 X 1000 = 0.05253 Gm. KNaC 4H 40 6
The U.S.P. requires that potassium and sodium tartrate contain not less than 99 per cent of KN aC 4H 40 S Calculate the
purity of the sample analyzed and compare the result with the
U.S.P. requirement.
1. Why cannot the alkali salts of such acids as citric, tartaric, etc., be
titrated directly with a strong acid, in the same manner as soluble carbonates
are titrated?
2. How much,of each of the following salts is equivaldnt to 1 cc. of 0.5
N H,.sO.? (a) KHC.H.O s, (b) K aC sH.0 7.H 2 0, (c) CH,COONa, (d)
LisC GH,07. 4H 20.
3. Name a number of official substances assayed by the above method.
Exercise 19
Object.-Assay of Magnesia Magma.

Materials Required.-5 Gm: of magnesia magma.
Procedure.-" After thorough agitation, place about 5 Gm. of Magnesia
Magma in a tared flask, stopper, and weigh accurately, add 25 cc. of normal
sulfuric acid, and after solution is complete, titrate the excess of acid with
normal sodium hydroxide, using methyl red T.S. as the indicator. Eaoh
cubic. centimeter of normal sulfuric acid is equivalent to 0.02917 Gm. of
Mg(OH)2,"
The magnesia magma is dissolved in an accurately measured

excess of normal sulfuric acid solution to insure complete neutralization of all of the magnesium hydroxide with the formation
ALKALIMETRY
95
of the soluble magnesium sulfate. The excess acid is then determined by residual titration with normal sodium hydroxide, using
methyl orange as 'indicator.
The reactions which take place when the magnesia magma is
dissolved and upon residual titration are as follows:
'Mg(OHh + H 2SOc---+MgS0 4 + 2H 20
58.34
H 2S0 4 + 2NaOH~Na2S04 + 2H 20
Each cubic centimeter of N H 2S0 4 consumed by the magma is
equivalent to:
58.34
2 X 1,000 = 0.02917 Gm. Mg(OH)2
If a sample of magnesia magma weighing 5.2430 Gm. when
dissolved in 25 cc. of 0.9915 N H 2S0 4 required 9.85 cc. of 1.1402
N NaOH to titrate the excess acid, the per cent Mg(OH)2 in
the sample could be calculated from the following formula:
[Ccc. H 2S0 4 X N) - (cc. NaOH X N)] X equivalent
Wt. of sample
X 100 = per cent, or substituting the figures from the example

cited above,
[(25 X 0.9915) - (9.85 X 1.1402)] X 0.02917 X 100 =
5.2430
.
7.54 per cent.
The U.S.P. requires that magnesia magma contain not less than
7 and not more than 8.5 per cent Mg(OH)2. The above sample,
therefore, conforms to the official requirements.
1. Why is residual titration resorted to in this assay?
2. Write all reactions ionically.
3. To how much Mg(OH). is 1 cc. of 0.1054 N HCI equivalent?
4. 12.32 Gm. of magma magnesia dissolved in 50 cc. of 1.0340 N H 2S0 4
required 24.60 cc. of 1.1255 N NaOH for residual titration. Calculate the
percentage Mg(OH)z present in the sample. To how much MgO does the
result correspond?
Exercise' 20
Object.-Assay of Methenamine.
96
Materials Required.-1 Gm. of methenamine.

Procedure.-1. "Place about 1 Gm. of Methenamine, accurately weighed,
in a beaker, add 40 cc. of normal sulfuric acid, and evaporate on a water
bath (or boil gently adding a little distilled water from time to time, if
necessary) until the odor of formaldehyde is no longer perceptible."
The weighed sample of methenamine is heated on a water

bath with a measured excess of normal sulfuric acid to decompose it into ammonia and formaldehyde, distilled water being
added from time to time to replace the water lost by evaporation.
Completion of the decomposition is indicated when the odor of
formaldehyde is no longer perceptible. The decomposition
reaction may be indicated in two steps as follows:
(CH2)6N 4 + 10H20~6CH20 + 4NH 40H
140.18
4NH40H + 2H2S04~2(NH4)2S04 + 4H 20
2. "Cool, add 20 cc. of distilled water, and titrate the excess of acid with
normal sodium hydroxide, using methyl red T.S. as indicator. Each
cubic centimeter of normal sulfuric acid correspon<!. to 0.03503 Gm. of
(CH2)6N 4."
The excess sulfuric acid found by residual titration with N

NaOH subtracted from the amount originally added gives the
amount of acid which combined with the liberated ammonia.
Since each mole of methenamine requires 2 mole~ of sulfuric
acid in the reaction, the quantity of methenamine equivalent to
1 cc. N H zS0 4 is
140.13
2 X 2 X 1,000 = 0.03503 Gm.
Calculate the perc.entage purity of the sample assayed.
1. Why should not the solution of ammonium sulfate be evaporated to
dryness to volatilize all of the formaldehyde in this assay?
2. Why is sulfuric acid a better acid to use than hydrochloric acid when
high temperatures are involved in the assay process?
3. If 1.12 Gm. of methenamine treated with 80 cc. of 0.5250 N H 2S0 4
require 12.40 cc. of 0.9885 N NaOH to titrate the residual acid, how pure is
the methenamine?
97
ALKALIMETRY
Exercise 21
Object.-Assay of Solution of Ammonium Acetate.

Materials Required.-25 cc. of solution of ammonium acetate.
Sodium hydroxide, T.S.
Normal sulfuric p,cid.
Procedure.-"Transfer 25 cc. of Solution of Ammonium Acetate to a
distilling flask, dilute with 75 cc. of distilled water, add 50 cc. of sodium
hydroxide T.S., and distil the liquid until all of the ammonia has been
driven over (about 100 cc. of distillate), receiving the distillate under the
surface of 50 cc. of normal sulfuric acid contained in a flask. Titrate the
excess of acid with normal sodium hydroxide, using methyl red T.S. as
the indicator. Each cubic centimeter of normal sulfuric acid is equivalent
to 0.07706 Gm. of CH,.COO.NH 4."
The NaOH reacts with the ammonium acetate liberating NHa

and combining with the acetic and carbonic acids to form nonvolatile compounds. The distilling flask should be connected
to the condenser at once after adding the NaOH to prevent loss
of the volatile NH a. The NHa formed in the reaction,
CH aCOONH 4
77.06
+ NaOH~CHaCOONa + NHa + H 0,
2
distils with water when the flask is heated. The NHa is conducted into a receiving flask which contains an excess of N
H 2S0 4 The condenser should be fitted tightly with an adapter
~n such a manner that the distillate will be collected beneath the
surface of ,the standard acid. This is done to prevent loss of
NHa from the distillate. As soon as the NHa in the distillate
comes into the receiving flask, it combines with the H 2S0 4 to
forni (NH 4 )S04,
2NHa
17.03
+ H2S04~(NH4)2S04
132.14
The excess acid is then determined by titration with standard

alkali.
"
1. Why is methyl red rather than phenolphthalein used in this assay?
2. Write equations for the reactions that occur in the assay of: (a)
aromatic spirit of ammonia, (b) ammonium bromide in tablets of three
bromides, (c) acid ammonium valerate, (d) ammonium carbonate.
98
3. Is this method of assay generally applicable to ammonium compounds?
Estimation of Nitrogen by the Gunning-Kjeldahl Method.Several methods have been developed for the so-called moist
combustion of nitrogen. The Gunning-Kjeldahl method which
will be described here, because it is the method specified in the
National Formulary for the estimation of the nitrogen content
of beef extract, illustrates the general principles involved in the
determination of nitrogen by all modifications of the Kjeldahl
method. See also peptone and casein, reagent, U.S.P.
The U.S.P. gives the following directions for the determination
of total nitrogen by the Kjeldahl method:
"Nitrates and Nitrites Absent.-Place about 1 Gm. of the substance, accurately weighed, in a 500-cc. Kjeldahl flask of hard
glass. The material to be tested, if solid or semi-solid, may be
wrapped in a sheet of nitrogen-free filter paper for convenience
in transferring it to the flask. Add 10 Gm of powdered potassium sulfate or anhydrous sodium sulfate, 0.5 Gm. of powdered
cupric sulfate, and 20 cc. of sulfuric acid. incline the flask at an
angle of about 45 and gently heat the mixture, keeping the
temperature below the boiling point of the mixture until frothing
has ceased. Increase the heat until the acid boils brliskly, and
continue the heating until the solution has been' clear green in
color for thirty minutes. Allow the mixture to cool, add 150 cc.
of distilled water, thoroughly mix the contents of the flask, and
cool again. Add cautiously 100 cc. of a 30 per cent aqueous
solution of sodium hydroxide added so as to cause the solution to
flow down the inner side of the flask to form a layer under the
acid solution. Add a few pieces of granulated zinc, connect the
flask, by means of a Kjeldahl connecting bulb, with a condenser,
the delivery tube from :which dips beneath the surface of a mixture
of 30 cc. of half-normal hydrochloric or sulfuric acid and 25 cc.
of distilled water contained in an Erlenmeyer flask or a widemouth bottle of about 500-cc. capacity. MiX; the contents of the
Kjeldahl flask by gentle-rotation, and distil until about two-thirds
of the contents of the flask has distilled over. Add about 5 drops
of methyl red T.S. to the contents of the receiving flask and determine the excess of acid by titration with half-normal sodium
hydroxide. Run a blank test and make necessary corrections.
ALKALIMETRY
99
Each cc. of half-normal acid consumed is equivalent to 0.007004

Gm. of nitrogen.
"When the nitrogen 'content of the substance is known to be
low, the half-normal hydrochloric or sulfuric acid may be replaced
by tenth-normal acid and tenth-normal alkali should then be used
in titrating the excess of acid. One cubic centimeter of tenthnormal hydrochloric or sulfuric acid is equivalent to 0.0014008
Gm. of nitrogen.
"With Nitrates Present.-Place a quantity of the substance,
accurately weighed, corresponding to about 0.15 Gm. of nitrogen,
FIG. 14.-A single-unit Kjeldahl apparatus.
in a 500-cc. Kjeldahl flask of hard glass, and add thereto 25 cc.

of sulfuric acid in which 10m. of salicylic acid has previously been
dissolved. Mix the contents of the flask thoroughly, and allow
the mixture to stand for thirty minutes with frequent shaking.
Add to the mixture 5 Gm. of powdered sodium thiosulfate and
again mix thoroughly, then add 0.5 Om. of powdered cupric sulfate and proceed as directed previously for Nitrates and Nitrites
Absent, beginning with 'Incline tfie flask at an angle of about 45.'
"NoTE.-There are certa~n alkaloids and other nitrogencontaining organic compounds that will not yield all of their
nitrogen to digestion with sulfuric acid, and this method, therefore, cannot be used for the determination of nitrogen in all
organic compounds."
100
The organic matter is decomposed by sulfuric acid with the

aid of a catalytic agent in a Kjeldahl apparatus (Fig. 14). The
carbon and hydrogen are converted into carbon dioxide and
water, respectively. The nitrogen is converted into an ammonium salt. The ammonia is liberated from this salt by a fixed
alkali and distilled into a measured quantity of acid of known
normality. The residual acid is then determined by titration
with a standard alkali solution, and the percentage of nitrogen
in the sample is calculated from the data obtained. This
method is applicable to the determination of nitrogen in most
organic and ammoniacal compounds but not to the estimation
of nitrogen in nitrates unless modified. It cannot be used for
the analysis of nitroso-, nitro-, azo-, and azoxy-compounds or
for certain other compounds such as pyridine and quinoline and
certain alkaloids, because these compounds do not yield all their
nitrogen when digested with sulfuric acid.
Exercise 22
Object.-To Determine the Amount of Nitrogen in the

Alcohol-soluble Solids of Extract of Beef.
Materials Required.-A Kjeldahl apparatus.
10 Gm. of extract of beef.
100 ce. of alcohol.
25 cc. of 0.1 N sulfuric acid.
25 cc. of 0.1 N sodium hydroxide.
20 cc. of concentrated sulfuric acid.
10 Gm. of potassium sulfate.
A 30 per cent solution of sodium hydroxide.
Cochineal or methyl red indicator solution.
All of the chemicals used must be of reagent quality and be

free from nitrates and ammonium salts.
Procedure.-l. Weigh about 10 Gm. of extract of beef accurately and
add sufficient distilled water to make 100 cc.
The extract of beef can be weighed conveniently in a small

evaporating dish. It may then be washed into a 100 cc. volumetric flask with distilled water.
2. Distribute 10 cc. of the solution over about 25 Gm. of previously
weighed sand or 5 Gm. of asbestos fiber contained in a previously weighed
fiat-bottomed porcelain dish and dried to constant weight in an oven at a
lOi
ALKALIMETRY
temperature of 1050. The weight of the residue from 10 cc. of the solution
corresponding to 1 Gm. of extract of beef should not be less than 0.75 dm.
corresponding to 75 per cent of total solids in the original extract.
The pasty mass of the extract is diluted and distributed over

sand or asbestos to expose a large surface area to the heat and
thereby hasten the evaporation of water. Extract of beef
containing less than 75 per cent of total solids does not meet the
National Formulary requirement.
3. Transfer another 25 cc. portion of the solution of beef extract to an
Erlenmeyer flask, add 50 cc. of alcohol, and shake the mixture vigorously.
When the precipitate which forms has subsided, collect it on a counterpoised filter paper, wash it three times with 5 cc. portions of a mixture
of 2 volumes of alcohol and 1 volume of distilled water, and then dry it to
constant weight at 1050. Reserve the filtrate and washings for the
determination of nitrogen.
The precipitate thrown down by the alcohol is considered to

be the alcohol-insoluble solids of beef extract.
4. Measure an aliquot portion of the alcoholic filtrate from the preceding determination corresponding to 1 Gm. of alcohol-soluble solids into
a 500 cc. Kjeldahl flask.
The aliquot portion corresponding to 1 Gm. of alcohol-soluble

solids of beef extract is calculated as follows: If the total solids
obtained from 10 cc. of the -solution containing 1 Gm. of the
extract of beef weigh 0.8 Gm., the percentage of total solids
in the extract will be 80 per cent. If the alcohol-insoluble solids
from 25 cc. of the solution containing 2.5 Gm. of beef extract
weigh 0.2 Gm., the percentage of alcohol-insoluble solids in the
total solids will be 2.5 0: . X 100 = 10 per cent. Each gram of
80
the extract must therefore correspond to 0.8 Gm. of total solids
and to 0.8 - 0.08 = 0.72 Gm. of alcohol-soluble solids. The
volume of the solution of beef extract corresponding to 1 Gm.
of alcohol-soluble solids in this c:i'se will be
0.~2
X 10 = 13.9 cc.,
where 10 is the dilution factor.

5. Add about 10 Gm. of powdered potassium sulfate and 20 cc. of concentrated sulfuric acid to the solution contained in the flask. Place the
flask in a slightly inclined position on an asbestos support with a circular
"102
opening so that the free flame will come in direct contact with the bottom
of the flask. Heat below the boiling point for about 10 min. or until frothing
has ceased and then raise the temperature and boil the mixture until the
latter acquires a pale straw color or is nearly colorless.
The proteins and other organic matter are oxidized by the

sulfuric acid-potassium sulfate mixture, leaving the nitrogen
combined as ammonium sulfate. If foaming occurs at first,
it can be prevented, for the most part, by dropping a small piece
of paraffin into the flask. The time necessary for the oxidation
of the compounds depends upon their nature, from Y2 t<J 12 hr.
heating being required by different compounds and mixtures.
A small crystal of copper sulfate or a globule of mercury may be
added to the digestion mixture to catalyze the oxidation.
6. Cool the flask, add about 250 cc. of distilled water, and cautiously
add a 30 per cent solution of sodium hydroxide until the contents of the
flask are distinctly alkaline. Use phenolphthalein added to the mixture
as indicator. Connect the flask at once to the Kjeldahl trap, condenser,
and receiver as illustrated in Fig. 14, so that the lower end of the condenser
dips beneath the surface of 25 cc. of 0.1 N sulfuric acid contained in the
receiving flask. Distil the mixture until about 100 cc. of distillate is
obtained.
In making the mixture alkaline, it should be borne in mind

that the pink, alkaline color of the phenolphthalein is destroyed
by a large excess of alkali. A few pieces of granutar zinc or
porous plate placed in the flask prior to distillation will prevent
bumping. The alkali liberates the ammonia from the ammonium
sulfate, and upon distillation the ammonia gas is driven over and
collected in the standard acid. In the analysis of compounds
containing a high percentage of nitrogen, all of the standard acid
may be neutralized before the whole of the ammonia has distilled.
If from 2 to 3 drops of methyl red or cochineal indicator are
added to the standard acid before the distillation is started, the
color of the indicator will show whether all the acid has been
neutralized, and in case it has, more standard acid can be placed
in the receiver without an appreciable loss of ammonia. The
Kjeldahl trap in the apparatus prevents liquid from the distillation flask from being carried over mechanically.
7. Add methyl red or cochineal indicator solution to the distillate and
titrate the excess acid with 0.1 N sodium hydroxide solution. Calculate
the percentage of nitrogen in the alcohol-soluble solids.
103
ALKALIMETRY
Deduct the number of cubic centimeter of 0.1 N alkali required

to neutralize the excess acid from the number of cubic centimeters of 0.1 N acid used. The difference will be the number of
TABLE IX.-OFFICIAL ALKALIMETRIC ASSAYS BY RESIDUAL TITRATION
Amount
used,
Gm. or
Substance
cc.
Nor- Equivamality lent of

of acid
1 cc.
Indicator
Official requirements,
per cent
--- --_ - - U.S.P.

Alnmonia, aroma-
tic spirit of (for

total NH,) ...
Ammonium
Ammonium carbonate ... , ... ,
Effervescent powders, compound
Glyceryl
trinitrate, tablets of.
Magnesia magma.
Magnesium
M.R.
0.5
0.008515 NH,
M.R.
1.0
0.07706
CR,COO.NR.
M.O.
1.0
0.01703
NH,
Pp.
0.5
0.04201
0.05 a
5
M.R.
M.R.
0.02 0.001514
1.0 0.02917
1
0.5
M.O.
M.O.
1.0
1.0
0.02016
0.02016
0.5
1
2
M.O.
M.R.
M.O.
1.0
1.0
0.5
0.02016
0.03503
0.04906
2
2
2
1.5
1.5
1
M.O.
M.O.
M.O.
M.O.
M.R.
M.R.
0.5
0.5
0.5
0.5
1.0
0.1
0.05254
0.05106
0.04102
0.04301
0.04069
0.004069
car-
bonate ........
Magnesium oxide.
Magnesium oxide,
heavy .........
Methenamine .. . ,
Potassium acetate
Potassium and so-
dium tartrate ..
Potassium citrate.
Sodium acetate .. .
Sodi urn ci tm te ...
Zinc oxide . . , ... ,

Zinc stearate ..
N.F.
..
= 1.7 to 2.1
10
25
ace-
tate, solution of
= 6.5 to 7.5 W IV
= 30-32
NaHCO, = 23 to 27
C,H,(NO,)' = 87.5 to 112.5'
Mg(OH), = 7 to 8.5
MgO = 39.2 to 41.5
MgO = 96
MgO = 96
(CH,),N. = 99
CR,COOK = 99
KNaC.H.O, = 99
K,C.H,O,.H,O = 99
CH,COON" = 99
Na,C,H,O, = 99
ZnO = 99
ZnO = 13 - 15.5
Ammonium bro-
mide in tablets
of three bro-
= 30.8 to
0.6
M.R.
0.5
0.04898
NH.Br
..
1.5
M.R.
0.5
0.05955
C.H.COONH. = 62 to 67
pared .......
Iron and ammon~
ium
acetate.
solution of. ....
Ichthammol (for
NR,) .......
Lithium carbo~~
ate ............
Lithium citrate ...
Methenamine,
ampuls of ......
Methenamine,
tablets of. .....
Methenamine and
sodium biphosphate, tablets
of .............
Zinc oxide in mild
resorcinol paste
Zinc oxide in
strong
resorcinol paste ..
1.5
M.O.
1.0
0.04069
ZnO
25
M.R.
0.5
M.R.
0.5
1.5
2
M.O.
M.O.
1.0
0.5
= 0.6 to 0.8W IV
= 2.5
0.03694 Li,CO, = 99
0.03498
C,H . OH.(COOLJla
M.R.
1.0
0.03503
(CR,),N.
M.R.
1.0
0.03503
(CH,),H.
mides ...... ,.
Ammonium valer-
ate, acid
Calamin.~.
Iwe-
35.8'
= 98
0.008515 NH.
0.008515 NR,
= 95
= 94
= 98.5
to 105'
to 106'
\"
= 92.5 to
= 24 to 26
0.25
M.R.
0.5
0.01752
(CH,),N.
M.O.
1.0
0.04069
ZnO
M.O.
1.0
o 04069
ZnO = 19 to 21
M.O. = methyl orange. M.R.

methyl red.
Pp.
phenolphthalein .
107.5'
104
cubic centimeters of the 0.1 N acid neutralized by the ammonia.

Since each cubic centimeter of 0.1 N acid neutralized by the
ammonia is equivalent to 0.001401 Gm. of nitrogen, the amount
of nitrogen obtained can be found by simple multiplication of the
number of cubic centimeters of 0.1 N acid neutralized by this
factor.
Quef'tions and Problems
1. Na!lle several nitrogen-containing products used in pharmacy to which
the Gunning-Kjeldahl method of analysis might be applied.
2. A nitrogenous substance weighing 6.150 Gm. furnished enough
ammonia, when analyzed by the Gunning-Kjeldahl method, to react with
40.65 ce. of 0.1 N H 2S0 4 What percentage of nitrogen did the substance
contain?
3. The nitrogen in a 5.0 Gm. sample of a food product was determined
by the Gunning-Kjeldahl method. After passing the evolved ammonia
into 90 cc. of 0.5 N acid, 25.80 cc. of 0.1 N alkali was required to titrate the
excess acid. 'Calculate the percentage of nitrogen in the sample.
4. How does the procedure for the determination of total nitrogen differ
from that given above when nitrates are present?
CHAPTER VI
ACIDIMETRY
Acids are estimated quantitatively by methods analogous to

those employed in alkalimetry, namely, by directly titrating an
exact quantity of the acid or acid salt with standard alkali
solution or by adding an excess of the latter and determining the
amount in excess by. residual titration with standard acid solution: Direct titration is employed whenever practicable, since
it is easier and requires fewer burette readings.
In assaying acids, the quantity of acid to be taken should be
such that about 30 to 40 cc. of the alkali solution will be consumed. As a general principle, it is recommended that the
normality of the acid to be titrated should be approximately
the same as that of the titrating medium. Except when otherwise directed, the liquid to be titrated should be brought to room
temperature before titration, as many indicators give different
values at different temperatures. For most of the inorganic
acids, methyl orange, methyl red, phenolphthalein, or litmus
can be used as indicators, but the alkali must be standardized
with the particular indicator used. For organic acids, phenolphthalein is always used.
DIRECT TITRATION METHODS
Exercise 23
Object.-Assay of Hydrochloric Acid.

Materials Required.-3 cc. of hydrochloric acid.
Procedure.-"Weigh accurately about 3 cc. of Hydrochloric ACHl III a
tared, glass-stoppered flask. Dilute with about 25 cc. of distilled water
and titrate with normal sodium hydroxide, using methyl red T.S. as the
indicator. Each cubic centimeter of normal sodium hydroxide is equivalent
to 0.03647 Gm. of He!."
Concentrated acid solutions are weighed in glass-stoppered

flasks to prevent loss of dissolved gases in some cases and to
105
106
QUANTITATIVE PHARMA(;1!Ju'1'lI.-'AL CHEMISTRY
prevent the absorption of water in other cases. Samples are

taken by weight rather than by volume, because it is difficult to
measure small volumes of concentrated acids accurately, and a
small error in the measurement of the sample would cause a
large per cent error in the result.
Hel + NaOH~NaCI
36.47
40.00
+H 0
2
The percentage of absolute HCI in the sample may be calculated:

cc. N NaQH X equivalent of HCI X 100
t
Wt. of sample of HCI
= per cen
The U.S.P. defines hydrochloric acid as an aqueous solution
containing not less than 35 and not more than 37 per cent of
HCI. Compare the strength of the acid assayed with the
official requirement.
1. A sample of HCI weighing 3.0024 Gm. required 34.45 cc. of 1.1542 N
NaOH upon titration. What per cent of HCI did the sample contain?
What is the Na 2CO. titer of the acid? What is the n:'6i'mality of the acid?
What is the molarity of the acid?
.
2. Calculate the per cent of chlorine in the sample in probl~m 1.
3. One cubic centimeter of a solution of HCI was found to be equivalent
to 1.12 cc. of NaOH solution. When the HCI was subsequently standardized, its HCI titer was found tODe 0.003642. What is the normality of the
NaOH solution?
,
4. A solution of HCI has a sodium carbonate titer of 0.0530. What weight
of sodium carbonate must be taken as a sample in order that the volume of
acid in cubic centimeters may give the per cent of Na 2C0 3 ?
Exercise 24
Object.-Assay of Diluted Sulfuric Acid.

Materials Required.-l0 cc. of diluted sulfuric acid.
Procedure.-" Accurately measure 10 cc. of Diluted Sulfuric Acid and
dilute with about 20 cc. of distilled water. Titrate the solution with normal
sodium hydroxide, using methyl red T.S. as the indicator. Each cubic
centimeter of normal sodium hydroxide is equivalent to 0.04904 Gm. of
H 2S0 4."
The official diluted acids are assayed, as a rule, by measuring

an exact quantity of the acid, titrating it after dilution, and
107
ACIDIMETRY
calculating the per cent of acid on a weight to volume basis.

This method of sampling is more convenient and rapid than
the taking of weighed' samples, and, since the volume of diluted
acid used as sample is relatively large, the error in measuring is
slight. The sulfuric acid reacts with the sodium hydroxide as
indicateq by the following equation:
H 2S0 4
98.08
+ 2NaOH--tNa S04 + 2H 0
2
2(40.00)
Since sulfuric acid is a dibasic acid and each mole is equivalent

to 2 moles of the monoacidic base, NaOH, the H 2S0 4 equivalent
of 1 cc. of 1 N NaOH will be 98.08/2 X 1,000 = 0.04904. Calculate the per cent of H 2S0 4 in the sample assayed.
1. What indicator solutions are suitable for use in the assay of diluted
sulfuric acid?
2. List a number of other official diluted acids, which are assayed by a
similar procedure, with the indicators used in each.
3. If the sample of H 2S0 4 assayed in this exercise had a specific gravity of
1.0640, what would be the per cent by weight?
4. Calculate the sodium carbonate and ammonium hydroxide titers of the
acid assayed in this exercise.
Exercise 26
Object.-Assay of Boric Acid.

Materials Required.-2.5 Gm. of boric acid.
100 cc. of glycerin.
Procedure.-" Dry about 2 Gm. of Boric Acid to constant weight over
sulfuric acid, weigh accurately, and dissolve the dried Acid in 100 cc. of a
mixture of equal volumes of glycerin and distilled water, previously neutralized to phenolphthalein T.S. Titrate with normal sodium hy_droxide, using
phenolphthalein T.S. as the indicator. Discharge the pink color by the
addition of 50 cc. of glycerin, neutralized to phenolphthalein T.S., and
again titrate until the pink color reappears. Each cubic centimeter of
normal sodium hydroxide is equivalent to 0.06184 Gm. of HaBO . "
If an aqueous solution of boric acid is titrated with standard

alkali solution using phenolphthalein as indicator, the end point
as shown by the development of the pink color appears before
all of the boric acid has been neutralized. If sufficient glycerin
is added to the boric acid previous to the titration, however,
108
the end point is sharply defined, the boric acid behaving as a

monobasic acid. The action of glycerin may be explained by
the assumption that glyceryl borate (C aH a0 20H)B(OH), a
stronger acid than boric acid, is formed, thus preventing the
formation of sodium metaborate. The hydrolysis of the latter,
if present, would increase the [OH-] causing the end point of
titration to be reached before an equivalent quantity of N aOH
has been added. Glycerin is frequently acid in reaction and
should, therefore, be neutralized before addition to the sample
being analyzed:
HaBOa
61.84
Therefore,
+ NaOH-NaH BO a + H 0
1 cc. N NaOH =
~10~~
,
= 0.06184
Gm. boric acid.

1. Why is glycerin added in this assay?
2. Write the reaction structurally.
3. Calculate the per cent HaBOa and the per cent B in the sample assayed.
Exercise 26
Object.-Assay of Tablets of Sodium Salicylate.

Materials Required.-20 tablets of sodium salicylate.
Diluted hydrochloric acid.
About 100 cc. of ether.
2 cc. of neutral alcohol.
Ferric chloride, T.S.
0.1 N sodium hydroxide.
Procedure.-l. "Weigh not less than 20 of the Tablets, reduce them to a
fine powder without an appreciable loss, and transfer an aliquot portion,
equivalent to about 0.3 Gm. of sodium salicylate, to a separatory funnel.
Add 25 cc. of distilled water and a slight excess of diluted hydrochloric acid,
and completely extract the liberated salicylic acid with ether. Transfer
the ethereal solution to a suitable flask and distil off most of the ether, being
careful not to volatilize the salicylic acid, and allowing the last few cubic
centimeters to evaporate spontaneously."
The tablets are weighed before powdering so that the average

weight per tablet can be computed. When Hel is added, salicylic
acid is set free as indicated by the following equation:
109
ACIDIMETRY
Shake the aqueous liquid successively with 20 cc. portions

of ether. The salicylic acid, being more soluble in ether than in
water, partitions itself into the ether layer. Completion of the
extraction can be determined readily by withdrawing 1 cc. of
the aqueous liquid and adding 3 drops of ferric chloride, T.S.
If no violet color forms in the test liquid, the extraction may be
considered to be complete. If a violet color appears, continue
to extract the aqueous liquid with 20 cc. portions of ether until
a negative test is obtained. Upon evaporation of the combined
ethereal solutions, any dissolved HCI is volatilized. In dil;ltilling
off the last of the ether, care should be used so that the flask
will not be heated above lOODC. because salicylic acid sublimes
slowly at higher temperatures.
2. "Add 2 cc. of neutral alcohol and 15 cc. of distilled water to the residue,
and titrate with tenth-normal sodium hydroxide, using phenolphthalein T.S.
as the indicator.
"Each cubic centimeter of tenth-normal sodium hydroxide is equivalent
to 0.01600 Gm. of sodium salicylate, C.H . OH.COONa.
"N OTE: This assay is applicable to these Tablets when not coated.
Suitable modifications or another method may be necessary for assaying the
Tablets when coated."
Neutral alcohol is added to dissolve the salicylic acid which is

but slightly soluble in water. Alcohol is frequently acid in
reaction and sh6uld always be neutralized if necessary, when
I
used as solvent in neutralization analyses.
C aH 4(OH)COOH
138.05
+ NaOH~CaH40HCOONa + H 0
2
160.04
One cubic centimeter of 0.1 N NaOH is equivalent to

2
?o 1;8i~~00 = 0.013805 Gm. of salicylic acid

Questions Ilnd Problems
1. Write equations for the reactions that occur in this assay using structural formulae.
2. Ascertain the solubilities of salicylic acid in water and in ether. Explain
why the acid partitions itself largely into the ether layer.
3. What amounts of sodium salicylate might tablets labeled "5 grain
tablets of sodium salicylate" contain and meet the official tolerance
allowances?
110'
4. From a bottle labeled" 5 grain sodium salicylate tablets," 20 sample

tablets were removed and found to weigh 7.9246 Gm. A 0.3 Gm. sample
taken from the powdered tablets required 16.05 cc. of 0.1012 N NaOH.
Calculate the amount of sodium salicylate contained in each tablet and the
per cent deviation of the tablets from the labeled amount.
Exercise 27
Object.-Assay of Tartaric Acid.

Materials Required.-3 GIll. of tartaric acid.
Procedure.-"Place about 3 Gm. of Tartaric Acid in a tared, glassstoppered flask and weigh accurately. Dissolve the Acid in about 40 cc.
of recently boiled distilled water, and titrate with normal sodium hydroxide,
using phenolphthalein T.S. as the indicator. Each cubic centimeter of
normal sodium hydroxide is equivalent to 0.07503 Gm. of H 2C 4H 40 6."
Phenolphthalein is the most suitable indicator to use in the

titration of organic acids. Tartaric acid reacts as a dibasic acid
as follows:
C 2H 2(OH)2(COOHh
150.05
+ 2NaOH---Na2C4H406 + 2H 0
2
Since 2 moles of N aOH are required to neutralize each mole of

tartaric acid, each cubic centimeter of N N aOH is \equivalent to
2 l~Oi~~o = 0.07503 Gm. tartaric acid.
1. Why is phenolphthalein employed as indicator in the titration of
organic acids with strong alkalies?
2. Write equations for the above reaction structurally.
3. How much citric acid is equivalent to 1 cc. of 1.0250 N NaOH? Indicate all calculations.
4. Write equations for the reactions that occur in the assay of; (a) acetic
anhydride, (b) trichloroacetic 'acid, (c) sulfosalicylic acid, (d) sodium bitartrate,
6. What would be the titer of each of the above substances for 0.1256 N
NaOH?

Exercise 28
Object.-Assay of Aromatic Sulfuric Acid.

Materials Required.-5 cc. of aromatic sulfuric acid.
'Ill
ACIDIMETRY

Procedure.-" Accura'tely measure 5 cc. of Aromatic Sulfuric Acid into a
tall beaker. Add exactly 30 cc. of normal sodium hydroxide and evaporate
the mixture to complete dryness on a sand bath. Dissolve the residue in
30 cc. of distilled water, and titrate the excess of alkali with normal sulfuric
acid, using methyl orange T.S. as the indicator. Each cubic centimeter of
normal sodium hydroxide is equivalent to 0.04904 Gm. of H 2S0 4."
Aromatic sulfuric acid contains ethyl sulfuric acid, alcohol,

and volatile oils in addition to sulfuric acid. The mixture
after treatment with a measured excess of N NaOH is evaporated
to complete dryness on a sand bath to decompose any ethyl
sulfuric aC,id and to drive off the volatile ingredients. The free
alkali and sodium sulfate which remain in the residue are then
dissolved in distilled water and the excess alkali is determined
by titration with normal sulfuric acid, using methyl orange
indicator, The small quantities of alkali consumed by the
.organic acids present in the volatile oil are negligible, since the
U.S.P. requirement is sufficiently broad to cover variation from
thiEl source, being not less than 19 per cent and not more than
21 per cent of H 2S0 4.
H 2S0 4 + 2NaOH~Na2S04 + 2H 20
98.08
Since 1 mole of H 2S0 4 requires 2 moles of NaOH, each cubic
centimeter of N NaOH is equivalent to 2
~~goo = 0.04904 Gm.
H 2S0 4
1. Why is methyl orange used in place of phenolphthalein in this assay?
2. Why is the solution of aromatic sulfuric acid evaporated to dryness on

a sand bath? Could a water bath be used?
3. Enumerate other offi'cial acidimetric assays in which residual titration
is employed.
.
Exercise 29
Object.-Assay of Tablets of Acetylsalicylic Acid.

Materials Required.-20 tablets of acetylsalicylic acid. (NOTE.-Two
5 grain tablets may be used for student work provided that the instructor
gives the weight of 20 tablets.)
112
oooo~o~ooooooooo~~~oooooo
~~~~o~o~~~~~~~~~ooo~~~~~~
ooo~~ooooooooooo~o~oooooo
~~~~o~~~~o~~~~~oo~o~o~~~~
cq
~
.....
1""'1
113
ACIDIMETRY
&<3.
;:::~
.s.
....
"''''
II II
~~
00
00
00
00
iii'S
88
~~
00
.....t~(O~
...-I
1""'4
a;llCc:v:I~I""'i"'d400
COlt:lQOOOO""lf400
..... ...-IC'-IcqOO....-l~'I""'I
0 0 0 0 ..... 0
..... 0
00000000
.....
00000
........
........
... 0
g~
~g
.......
00
00
00
00
00
;o..~
~~
~~
z'S
..... I""'iP"'ll"""4d
.... O,....4
0000""';0"';0
---------7~--------------------------------~~
1.r,)lt:)lt:)COOOOl
oooOcO"';c-io
. 0. . .
It:)
It:)
00
0",
OLQOO",,",
'"
00
II
.""
a'"
.S
:"g
:o;S"
: .~
s
:~
"'"
114
QUANTITATIVE PHARMACEUTICAL CHEMISTR
Procedure.-l. "Weigh not less than 20 of the Tablets, H'Uuce them to a

fine powder without an appreciable loss, and dissolve <hn aliquot portion,
representing approximately 0.7 Gm. of acetylsalicylic acid, in 20 cc. of
neutral alcohol in an Erlenmeyer flask, and titrate the1solution immediately
with tenth-normal sodium hydroxide, using phenolphthalein T.S. as the
indicator."
Care must be observed in powdering the tJ\blets because the

sample will not be representative if any of the,material is lost.
The sample is titrated with sodium hydroxide to neutralize any
free acid formed by hydrolysis of the acetyls4licylic acid as
represented by the equation:
C 6H 4 .OCOCH a.COOH
+ HOH--+C 6H 4 .OH.COOIi +
CHaCOOH
and to neutralize the carboxy group of the acetylsalicylic acid:

C 6H 4 .OCOCHa.COOH
180.06
+ NaOH~06H4.000CH3CO(j)Na +
HOH
2. "Now add a volume of tenth-normal sodium hydroxide equal to that

used in the above titration plus 5 ce. more. Heat the mixture on a steam
TABLE XL-OFFICIAL ACIDIMETRIC ASSAYS BY RESIDUAL TITRATION
Amount
used,
Gm. or
cc .
Substance
U.S.P.
Acid, acetylsalicylic
Acid, lactic ........
Acid, sulfuric aromatic ...........
Chloral hydrate ...
Formaldehyde,
solution ....... .
Methyl salicylate ..
N.F.
Acetylsalicylic acid,
tablets of. ......
Ethyl acetate .....
Nor- Equivamality lent of

of alkali 1 cc.
Indicator
I
--- - _ -
1.5
2.5
Pp.
Pp.
0.5
1.0
0.04502
0.09005
C.H.O(CH,CO)COOH = 99.5
CH,CHOHCOOH = 8S,to 90
S.O
4.0
M.O.
Pp.
1.0
1.0
0.04904
0.1654
H,SO. = 19 to 21 W IV
CChCHO.H,O = 99.5
3.0
2
B.T.B.
Pp.
1.0
0.5
0.03002
0.07603
HCHO = 37
C.H . OH.CO,CH, = 98
O.;7a
Pp.
0.1
0.01801
I.S
Pp.
0.5
0.04403
C.H . OCOCH,.COOH =:92.5

to 107.5'
CH,COO.C,H. = 99
Pp.
0.1
0.004602 HCOOH
Formic acid, spirit
of. ..
.. ,
"
10
B.T.B. = bromthymol blue. M.O.

, Per cent of labeled amount.
methyl orange.
l'p.
0.95 to 1.0SWIV
phenolphthalein
115
ACIDIMETRY
bath for 15 minUtes, with intermittent stirring, allow the solution to stand
at room temperatu~e for 5 minutes, and then titrate with tenth-normal
sulfuric acid. Subtract the number of cubic centimeters of tenth-normal
sulfuric acid used, frOm the total number of cubic centimeters of alkali
added the second time, and multiply the result by 0.01801. This result
represents the weight of acetylsalicylic acid present in the portion of the
pow.der used for the a~say."
Upon heating ,the titration mixture after the addition {)f an

excess of 0.1 N a,lkali, the ester is saponified:
C 6li4.0COCH 3.pOONa
+ NaOH~C6H4.0H.COONa +
CH3 COONa
+ HOR
The amourrt of 0.1 N alkali remaining after the saponification

is complete is then determined by titration with 0.1 N acid.
I,. Show,how the equivalent for acetylsalicylic acid is derived.
CHAPTER VII
PRECIPITATION METHbD~
Under Neutralization Methods, a class of re~ctions was considered which were of value in quantitative analysis because little
ionized substances or gases or both little ionized substances and
gases were formed. In the following exercises, a cla;;s of reactions
is dealt with which requires the formation of very slightly
soluble substances to cause the reactions to go to sufficient completion to be quantitative in nature. The solubility product
principle may be applied to all precipitation reactions, and it
should be reviewed before the subsequent assays are undertaken.
Determination of the End Point.-The end point of a reaction
in analyses by precipitation methods may be determined in one
of three ways:
1. By adding a standard solution to a solution of the substance
being analyzed until no further precipitate is prbduced. This
method is sometimes applied in the determination of the chloride
ion content of chlorides with standard solution of silver nitrate.
2. By adding the standard solution to a clear solution of the
substance being analyzed until a precipitate begins to form:
This method is often used in the titration of alkali cyanides with
standard silver nitrate solution. (See sodium cyanide reagent,
U.S.P. and diluted hydrocyanic acid, N.F.)
3. By means of a suitable indicator. All of the official assays
by precipitation methods employ indicator solutions to show the
end point of the reaction.
Indicators.-The indicators used in the official volumetric
precipitation assays are:
1. Ferric ammonium sUlfate T.S. prepared by dissolving 8
Gm. of reagent ferric ammonium sulfate in sufficient distilled
water to make 100 cc. This indicator is used in both direct and
residual tit rations of silver and mercury salts with standard
ammonium thiocyanate solution. The thiocyanate reacts with
116
PRECIPITATION METHODS
117
the silver or mercury present to form a white precipitate of

silver or mercury thiocyanate but as soon as all of the silver
or mercury has been precipitated the thiocyanate reacts with
ferric ammonium sulfate to form red ferric sulfocyanate; the
first appearance of the red color marks the end point of the
reactlon.
2. Potassium chromate T.S. prepared by dissolving 10 Gm. of
reagent potassium chromate in sufficient distilled water to make
100 cc. It may be used as indicator in the titration of chloride
solutions with standard silver nitrate solution, but in the Pharmacopoeia it is employed only in the assay of oil of bitter almonds
for hydrocyanic acid. The end point is shown by the production
of a permanent red color caused by the formation of silver
chromate.
Standard Solutions.-The standard solutions employed in
the official assays by precipitation methods are tenth-normal
silver nitrate and tenth-normal ammonium thiocyanate.
Exercise 30
Object.-To Prepare and Standardize 0.1 N Silver Nitrate.

Materials Required.-lO Gm. of silver nitrate.
Procedure.-Dryabout 10 Gm. of pure silver nitrate in an electric oven
at 1100. until of constant weight. Weigh out exactly 8.4945 Gm. of the
dried salt and dissolve it in sufficient distilled water to make 500 cc.
The pure analyzed silver nitrate now available on the market

makes it possible to prepare the solution directly from the
dried salt. Every precaution should be exercised in weigqing
the silver nitrate, since a slight error in the weight would make
the concentration of the solution inaccurate:
Mol. wt. AgNO s = 169.89
Therefore 1,000 cc. of 0.1 N AgNO s requires 169.89/10 =
16.989 Gm., and 500 cc. of 0.1 N AgNO a requires 16.989/2 =
8.4945 Gm. pure AgNO a \.
If the purity of the silver nitrate crystals used is doubtful, the
concentration of the volumetric solution may be determined by
titration against an accurately weighed sample of pure sodium
chloride in the same manner as given under the assay of sodium
chloride.
118
The distilled water should be tested for halides as follows

before it is used to prepare standard silver nitrate solution:
Fill a test tube three-fourths full of water, add about 0.5 cc.
of silver nitrate solution and a few drops of U.S.P. nitric acid.
Any turbidity is positive evidence that the water contains
halides and is unfit for use in preparing the standard soiution
unless the concentration of the solution is to be ~etern;l.ined by
titration against a pure chloride.
Exercise 31
Object.-To Prepare and Standardize 0.1 N Ammonium

Thiocyanate.
Materials Required.-4.0 Gm. of ammonium thiocyanate.

0.1 N silver nitrate.
""'" in 500
Procedure.-Dissolve 4.0 Gm. of reagent ammonium thiocyanate
cc. of distilled water. Ascertain its strength by titration against tenthnormal silver nitrate, and adjust to exact normality or to a known strength
of approximate normality.
Since ammonium thiocyanate is a deliquesJlent salt, a slight

amount in excess of the theoretical quantity required is u~d to
prepare the solution. The normality of the soluti~n is determined by titration with 0.1 N AgNO a as follows:
Measure exactly about 30 cc. of 0.1 N AgNO a into a 250 cc.
Erlenmeyer flask, and dilute the solution with 50 cc. of distilled water, add 2 cc. of ferric ammonium sulfate T.S. and
2 cc. of nitric acid to prevent hydrolysis of the ferric alum,
and titrate the mixture with the approximately 0.1 N NH 4SCN
solution. When the thiocyanate solution is run in, a white
precipitate is produced giving the mixture a milky appearance,
and as the thiocyanate is added drop by drop, it produces a
brownish-red cloud which quickly disappears on shaking. As the
end point is reached, the precipitate becomes flocculent settling
readily and, finally, a drop or two of thiocyanate solution produces a permanent reddish color which does not disappear on
shaking.
Ammonium thiocyanate solution added to an acidulated solution of silver nitrate containing ferric alum precipitates silver
thiocyanate until all of the silver has been precipitated, then the
119
slightest excess of ammonium thiocyanate solution reacts with

ferric alum to form red ferric thiocyanate as follows:
I
AgNO a + NH 4SCN--+AgSCN + NH 4N0 3

169.89
76.11
white
2FeNH 4(S04)2 + 6NH 4SCN--+2Fe(SCN)a + 4(NH 4)2S04
.
red
If 30.00 cc. of 0.1 N AgNO a require 28.13 cc. of NH 4SCN solution, the factor of the NH 4SCN solution in terms of 0.1 N is
30.00/28.13 = 1.0666.
Oxides of nitrogen yield colored salts with ferric alum. Solutions containing oxides of nitrogen, therefore, should be boiled
prior to the addition of the indicator.
1. Explain in detail how to standardize a solution of silver nitrate against
pure sodium chloride.
2. Why is nitric acid added when NH.SCN solution is standardized
against 0.1 N AgN0 3 using ferric alum as indicator?
3. Write ionically the reactions involved in the standardization of
NH.SCN against AgN0 3 with ferric alum indicator.
4. What must be the normality of solution of silver nitrate so that each
cubic centimeter shall be equivalent to 6 mg. of N aCI?
5. How might 0.1 N AgN03 be prepared from pure metallic silver?
6. Why should standard silver nitrate solution be protected from strong
sunlight?
7. Why must oxides of nitrogen be absent in titrations with NH.SCN?
8. How much 0.1 N AgN0 3 could be prepared from 5 Gm. of pure silver?
9. To hOw much Hg, Ag, toughened AgN0 3, and HgO is 1 cc. of 0.1000
N NH.SCN equivalent?
10. If 1 cc. of a solution of silver nitrate is equivalent to 0.0148 Gm. of
NaCI, what is the normality of the silver nitrate solution?
DIRECT TITRATION METHOD
Compounds of silver and mercury which can be readily converted into silver nitrate and mercuric nitrate, respectively,
may be estimated by direct titration with standard potassium
thiocyanate soluti6n, using ferric ammonium sulfate as indicator.
The method is based on the quantitative precipitation of the
corresponding thiocyanate, e.g.:
120
and
When all of the metallic ion has been precipitated as thiocyanate, the NH 4SCN reacts with the ferric alum indicator to
form red-colored ferric thiocyanate, marking the end point of
titration:
The solution must be acidulated with nitric acid to prevent the

hydrolysis which ferric salts undergo in neutral solution. Chlorides must be absent, since the chlorides of silver and mercury
are more soluble than the respective thiocyanates. This
method of assay is exemplified in the following exercise:
Exercise 32
Object.-Assay of Strong Silver Protein.

Materials Required.-2 Gm. of strong silver protein.
About 10 cc. of nitric acid.
2 cc. of ferric alum indicator (8 Gm. FeNH 4(S04)2.12H 20 in 100 cc.).
0.1 N ammonium thiocyanate.
I
Procedure.-l. "Ignite about 2 Gm. of Strong Silver Protein, accurately
weighed in a porcelain crucible until all of the carbon is burned off. Transfer as much as possible of the residue to a beaker, add to the crucible 5 ce. of
nitric acid, warm to dissolve any adhering silver, and transfer the solution
to the beaker with the aid of a little distilled water. Cover the beaker, and
heat on a water bath until all of the metallic silver is dissolved, adding a
little more nitric acid, if necessary."
Upon ignition, strong silver protein yields a residue of metallic

silver and silver oxide. The silver and silver oxide are dissolved
in nitric acid:
Ag
2HNO a----+AgNO a
H 20
N0 2
Ag 20
2HNO a----+2AgNO a H 20 .
2. "Filter into a flask, wash the insoluble residue thoroughly with distilled water, cool, and dilute with distilled water, if necessary, to about
75 cc. Add 2 cc. of ferric ammonium sulfate T.S., and titrate with tenthnormal ammonium thiocyanate Each cubic centimeter of tenth-normal
ammonium thiocyanate is equivalent to 0.01079 Gm. of silver."
121
The mixture is filtered to free it of carbonaceous matter,

the residue on the filter is washed free of silver nitrate, and the
nitrate is titrated wi'th 0.1 N NH 4SCN:
AgNO a NH 4SCN--AgSCN
NH 4 NO a
169.89
Each cubic centimeter of 0.1 N NH 4SCN is equivalent to
107.88
10 X 1000 = 0.01079 Gm. Ag. The U.S.P. requires that strong
silver protein contain 7.5 to 8.5 per cent silver. Calyulate the
percentage of silver in the sample analyzed and compare your
result with the U.S.P. requirement.
1. How does the solubility product principle apply in the above assay?
2. If each cubic centimeter of a solution of NH,SCN is known to be
TABLE XIl.-OFFICIAL SUBSTANCES ASSAYED BY DIRECT TITRATION WITH
0.1 N NH,SCN
Substance
Amount Equivaused, Gm. lent of

or cc.
1 cc.
0.01003
0.01083
0.01003
0.01003
0.01083
0.01003
0.01003
0.01003
0.01003
0.01699
0,01699
0.01079
0.01079
Hg = 32 to 34
HgO = 99.5
Hg = 54 to 59.5
Hg = 49.5 to 51
HgO = 24 to 26
Hg = 99.5
Hg = 37 to 39
Hg = 29 to 31
Hg = 49 to 51
AgNO. = 99.8
AgNO. = 94.5
Ag = 19 to 25
Ag = 7.5 to 8.5
0.02272
0.01083
0.01003
0.01003
HgI 2 = 0.95 to 1.05

HgO = 99.5
Hg = 51.0 to 62.4 b
Hg = 47.7 to 52.8b
U.S.P.
Mass of mercury. . . . . . . .. . .. .
Mercuric oxide, yellow ........ .
Mercuric salicylate ........... .
Mercuric succinimide ....... .
Mercury, oleate of. . . . . . . . .. ..
Mercury., ..... , ..... , ..... " ..
Mercury with chalk ... , , ... , . , .
Ointment, mercurial, mild .. , . , .
Ointment, mercurial, strong, ...
Silver ni-trate .... , .. , ..... , .. ,
Silver nitrate, toughened, .. , ...
Silver protein, mild. , ... , . , , .. ,
Silver protein, strong ... , .. , ...
0.5
0.5
0.5
0.5
0.75
0.4
1.0
1.0
1.0
0.8
0.8
1.0
2.0
N.F.
Arsenic and merouric iodides,.
solution of (for HgI 2 ) . 25
Mercuric oxide, red. . ..... , ...
0,5
Mercuric salicylate, ampules of
0.5 a
Mercuric succinimide, ampuls of I 0.25"
Amount of ingredient sought.

122
equivalent to 0.01140 Gm. of pure Ag, what would be its equivalent of Hg,
HgI HgO, Hg(NO.) AgNO., and Ag.O?
3. Give a method for the assay of metallic mercury. Explain each step
in the procedure and write equations for all reactions.
4. What must be the normality of a solution of NH 4SON so that each
cubic centimeter shall be equivalent to 1 mg. of silver?
o. Write equations for the reactions that occur in the assay of: (a) yellow
mercuric oxide, (b) mercuric succinimide, (c) mercuric iodide in solution of
arsenic and mercuric iodides.
RESIDUAL TITRATION METHOD (VOLHARD'S METHOD)
This method is based on the complete precipitation of insoluble

silver salts from nitric acid solution by the addition of .excess
standard silver nitrate solution to a soluble salt and the determination of the amount of silver nitrate solution in excess by
residual titration with standard ammonium thiocyanate solution,
using ferric ammonium sulfate as indicator.
Exercise 33
To Determine the Purity of a Soluble Chloride.-The purity of

any soluble chloride may be determined by the method given
below, provided that other substances which give precipitates
with silver nitrate are absent. Substances other than chlorides
which produce slightly soluble silver salts are bromides, iodides,
cyanides, sulfocyanides, sulfides, phosphates, arsenates, carbonates, etc.
Object.-Assay of Sodium Chloride.
Materials Required.-l Gm. of sodium chloride.
5 cc. of nitric acid.
2 cc. of ferric alum indicator.
50 cc. of 0.1 N silver nitrate.
Procedure.-" Dry about 0.25 Gm. of Sodium Ohlbnae to constant weight
at 1000., weigh accurately, and dissolve in 50 cc. of distilled water in a
200-cc. volumetric flask. Add 50 cc. of tenth-normal silver nitrate to the
solution, then 5 cc. of nitric acid, and sufficient distilled water to make .the
mixture measure 200 cc. Mix well, filter into a dry flask through a ffiter
which has not been moistened, and reject the first 20 cc. of ffitrate. To
exactly 100 cc. of the subsequent filtrate add 2 cc. of ferric ammonium sulfate
T.S., and titrate with tenth-normal ammonium thiocyanate. Each cubic
centimeter of tenth-n~rmal silver nitrate is equivalent to 0.005845. Gm. of
NaOl."
PRECIPlTATION METHODS
123
Nitric acid is added to prevent the precipitation of silver as

carbonate, phosphate, etc. and to prevent hydrolysis of the ferric
alum.
.
The precipitated chloride is removed by filtration, since
silver chloride is several times more soluble than silver thiocyanate. If the precipitated silver chloride were left in the
titration mixture when the excess silver nitrate is titrated with
ammonium thiocyanate, the following reaction would take place
between the dissolved AgCl and NH 4SCN:
AgCl
+ NH4SCN~AgSCN + NH4Cl
More precipitated AgCl would then dissolve to re-establish

eqvilibrium between the precipitate and solution. This would
make the end point uncertain.
The residual liquid after precipitation, therefore, is made up
to a definite volume, 200 cc., and an aliquot portion, 100 cc., of
the filtrate is taken for residual titration. The first 20 cc.
of filtrate is rejected to avoid possible error due to absorption of
silver nitrate by the filter. The aliquot-portion method eliminates the tedious procedure of washing the precipitate free of
silver nitrate which would be required if all of the excess silver
nitrate were to be titrated residually.
+ AgNOa~AgCl + NaNO a
AgNO a + NH4SCN~AgSCN + NH4NOa
NaCl
58.45
Each cubic centimeter of 0.1 N AgNO a is equivalent to

58.45
10 X 1000
= 0.005845 Gm. NaCl.
The number of cubic centimeters of 0.1 N NH 4SCN required in

the residual titration is multiplied by 2 because only pne-half the
excess 0.1 N AgNO a is represented in the aliquot portion.
[cc. 0.1 N AgNO a - (2 X cc. 0.1 N NH 4SCN)] X 0.005845
Wt. of s~mple
X 100 = per cent purity of NaCl
1. Why is nitric acid added in the above assay?
2. Why must the precipitated AgCl be removed before titration with
standard NH 4SCN? Explain in detail.
124
3. How might all of the excess silver nitrate solution be freed from the
precipitate and titrated?
4. Name several official chlorides assayed by the above method.
6. What must be the normality of a standard silver nitrate solution so
that each cubic centimeter will be equivalent to 1 mg. of chlorine?
Exercise 34
Object.-Assay of Ammonium Bromide.

Materials Required.-0.4 Gm. of ammonium bromide.
2 cc. of ferric alum indicator.
50 cc. of 0.1 N silver nitrate.
Procedure.-"Dry about 0.4 Gm. of Ammonium Bromide to constant
weight in a desiccator over sulfuric acid, and weigh accurately. Dissolve
it in about 50 cc. of distilled water, add 50 cc. of tenth-normal silver nitrate,
2 cc. of ferric ammonium sulfate T.S., and 2 cc. of nitric acid. Titrate the
excess of silver nitrate with tenth-normal ammonium thiocyanate. Each
cubic centimeter of tenth-normal silver nitrate is equivalent to 0.009796 Gm.
of NH 4Br. Each gram of Ammonium Bromide, previously dried, is equivalent to not less than 101.1 cc. and not more than 103.0 of tenth-normal
silver ni trate."
The above assay is similar to that for chlorides, except that the
precipitated silver bromide need not be removed by filtration,
since it is less soluble than silver thiocyanate. I
The reactions and calculations are similar to those of the preceding assay. Write equations for all reactions involved and
calculate the per cent purity of the ammonium bromide.
1. How does the presence of excess silver nitrate tend to produce quantitative precipitation of silver bromide?
2. How does the formation of insoiuble silver bromide force the reaction
to completion?
3. Why may the residual titration of excess silver nitrate be conducted
without removal 0{ the precipitated silver bromide?
4. Look up the solubility product of AgBr and calculate the solubility of
silver bromide.
5. Write all equations ionically.
Exercise 35
Object.-Assay of Syrup of Hydriodic Acid.

Materials Required.-25 cc. of syrup of hydriodic acid.
40 ce. of 0.1 N silver nitrate.
125

2 cc. of ferric alum indioator.
Procedure.-"Place exactly 25 cc. of Syrup of Hydriodic Acid in a flask,
dilute it with 100 cc. of distilled water, add 40 cc. of tenth-normal silver
nitrate, agitate the mixture, add 5 cc. of nitric acid and heat the mixture on
a water bath until the preoipitate has acquired a bright yellow color. Cool,
add 2 cc. of ferric ammonium sulfate T.S., and determine the residual silver
nitrate by titration with tenth-normal ammonium thiocyanate. Each cubic
centimeter of tenth-normal silver nitrate is equivalent to 0.01279 Gm. of HI."
This assay is similar to that for bromides except that a

measured volume of liquid is used for the sample.
HI + AgN03~AgI + HNO a
127.93
AgNO a + NH4SCN~AgSCN + NH 4NO a
Calculate the amount of HI equivalent to 1 cc. of 0.1 N AgNO a
and the per cent HI in the sample analyzed. The U.S.P.
requires that syrup of hydriodic acid contain not less than 1.3
and not more than 1.5 Gm. of HI in each 100 cc. Note that
this is a weight-to-volume percentage.
1. Why must a large amount of nitric acid be avoided in the precipitation
of chlorides, bromides, and iodides as the corresponding silver salts?
2. If 25 cc. of syrup of hydriodic acid containing 1.4 per cent HI is
treated with 40 cc. of 0.0955 N AgNO., how much 0.1225 N NH 4 SCN will be
necessary to titrate the excess AgNO.?
3. Write equations lor the reactions that occm in the assay of (l,) cbloroform liniment, (b) potassium nitrate, (c) syrup of ferrous iodide, and (d)
acriflavine.
4. A solution is known to contain 5.3490 Gm. of silver combined as nitrate
per 1,000 cc. How much of each of the following substances would be
equivalent to 1 cc.: Br, CaCh, FeI 2, HCI, NaI, NaCN, and NH.CI?
Exercise 36
Object.-Assay of Elixir
of Three Bromides ..
Materials Required.-10 cc. of elixir of three bromides.

0.1 N silver nitrate .
. 2 cc. of nitric acid.

2 cc. of ferric alum indioator.
126
Procedure.-"Dilute exactly 10 cc. of the Elixir with distilled water to

250 cc. To 25 cc. of the dilution, add slowly and with agitation, 50 cc. of
tenth-normal silver nitrate, 2 cc. of nitric acid, and 2 cc. of ferric ammonium
sulfate T.S. Titrate the excess silver nitrate with tenth-normal ammonium
thiocyanate.
Each cubic centimeter of tenth-normal silver nitrate is equivalent to
0.01059 Gm. of the total bromides."
In calculating the amount of bromide equivalent to 1 cc. of

0.1 N silver nitrate in elixir of three bromides, elixir of five
bromides and syrup of five bromides and similar mixtures the
following general formula may be used:
F
+ C2b + C3ba, etc.

Cl
bl
F = number of grams of mixed bromides equivalent

to 1 cc. 0.1 N AgN0 3
a = total number of grams of bromides (based on
100 per cent purity) in mixture.
bl , b2 , b3 , etc. = number of grams of individual bromides (100
per cent pure) in mixture.
CI, C2, C3, etc. = number of grams of the broniide represented by
bl , b2 , b3, etc., equal to 1 cc. 0.1 N AgN0 3
The factor for elixir of three bromides, which contains 80 Gm.
of sodium bromide, 80 Gm. of ammonium bromide and 80 Gm.
of potassium bromide per 1,000 cc. of solution, is I calculated
thus (assuming 100 per cent purity):
240
F =
80
80
80
= 0.OlO59
where
0.010291
+ 0.Oll901 + 0.009796
The factor for syrup of five bromides may be calculated as

follows: The syrup contains 80 Gm. of potassium bromide, 80 Gm.
of sodium bromide, 50 Gm. of ammonium bromide, 25 Gm. of
calcium bromide and 8 Gm. of lithium bromide per 1,000 cc.
of solution. If these substitutions are made in the formula,
237.8
= --O;:8=0----;:8:-;:;:0----c5;;ro;------;2::;-;;5;-:-x:-:0;:-.8:::-4-;---8~X-:--;:-0.-==85.
0.011902
+ 0.010292 + 0.009796 +
0.009996
+ 0.008686
= 0.01058
127
TABLE XIII.-OFFICIAL SUBSTANCES ASSAYED BY RESIDUAL TITRATION OF
0.1 N AgNO.
Amount
used.
Gm.
or co.
Substance
U.S.P.
Acid hydriodic. diluted ...... '.' ..
Acriflavine .................... .
Acriflavine hydrochloride ....... .
Ammonium bromide ....... ... .
Ammonium chloride .... .
Calcium chloride. R ........ .
Chloroform. liniment of. .. . ...
Chloroform. spirit of. . . . . . .
..
Oil of mustard. volatile. .. .. . .
Potassium bromide ............ .
Potassium nitrate .............. .
Sodium bromide ............... .
Sodium chloride ............... .
Syrup of ferrous iodide ......... .
Syrup of hydriodic acid ......... .
N.F.
Ammonium bromide, elixir of
.,
Ammonium chloride. tablets of.
Ammonium iodide ............ .
Bromides, five, elixir of . . , .. - ... .
Bromides, three, elixir of, ....... .
Bromides. three. tablets of. ..... .
Lithium bromide .............. .
Potassium broIuide. elixir of . .... .
Potassi urn chloride . ............ .
Potassium thiocyanate .. , ....... .
Sodium hromide. elixir of. ...... .
Sodium bromide. tablets of ...... .
Sodium chloride in isotonic solution of dextrose and. . ....... .
Sodium chloride in ampuls of dextrose and ................... .
Sodium chloride. ampuls of ...... .
Sodium nitrite. tablets of ....... .
Sodium thiocyanate ... ......... .
Strontium bromide ............. .

Zinc iodide .................... .
R.
a
reagent.
Amount of ingredient Bought.
Per cent of the.labeled amount.
=
WITH
0.1 N NH.SCN
Equivalent of
1 cc. of
0.1 N
NH,SCN
per cent
_--
5
0.25
0.25
0.4
0.1
2
10
5
4
0.4
0.4
0.4
0.25
10
25
0.01279
0.003546
0.003546
0.009796
0.005350
0.0055.50
0.00398
0.00398
0.004956
0.01190
0.01011
0.01029
0.005845
0.01548
0.01279
HI = 9.5 to 10.5
Cl = 13.3 to 14.3
Cl = 23 to 24.5
NH,Br = 99
NH,CI = 99.5
CaC]' = 74
CHCla = 40 to 45 W /V
CHCla = 8.5 to 9.25 W /V
C,H,NCS = 93
KBr = 99
KNO, .:, 99
NaBr = 99
NaCI = 99.5
FeI, = 6.5 to 7.5W/V
HI = 1.3 to 1.5W/V
10
0.15
0.5
10
10
0.6
0.35
5
0.25
0.2
5
0.3"
0.009796
0.00535
0.01450
0.0102
0.01059
0.007992
0.008686
0.01190
0.007455
0.009716
0.01029
0.01029
NH,Br = 8 to 9W/V
NH,CI = 92.5 to 107.5'
NH,I = 98
total bromides = 25 to 27 W /V
total bromides = 23 to 25 W /V
Br = 70 to 81 b
LiBr = 85 to 90
KBr = 17 to 18W/V
KCI = 99
KSCN = 99
NaBr = 17 to 18W/V
NaBr = 92.5 to 107.5b
0.2"
0.005845 NaCI
0.2
0.2"
0.5"
0.2
0.5
0.5
0.005845
0.005845
0.02070
0.008107
0.01778
0.01596
~
= 0.4 to 0.425W/V
NaCI = 95 to 105 b
NaCI = 95 to 105 b
NaNO, = 91 to 109b
NaSCN = 98.5
SrBro.6H,O = 98
ZnI, = 98
128
According to the U.S.P. XI, calcium bromide should contain

at least 84 per cent of CaBr2, thus we have to multiply 25 Gm. by
84 per cent to obtain the amount of 100 per cent CaBr2 present
The same applies to lithium bromide; the N.F. VI allows a
minimum of 85 per cent LiBr.
1. Show how the total bromide equivalent in the assay of elixir of five
bromides is derived.
2. Tablets of three bromides each weighing on the average 0.4252 Gm.
required 37.50 cc. of 0.1 N AgN0 3 in the assay of a powdered sample of the
tablets weighing 0.4012 Gm. Calculate the per cent of bromine and of total
bromides in the tablets.
3. Calculate the weight of bromine and of total bromides in each tablet
from the data given in problem 2.
4. If the tablets assayed as in problem 2 were labeled" 5 grain tablets of
three bromides," would they conform to the official requirements?
5. Calculate the minimum and maximum amounts of total bromides that
the tablets labeled as in question 4 might contain and meet the official
requirements.
CHAPTER VIII
OXIDATION -REDUCTION METHODS
The chemical reactions which occur in neutralization and

precipitation methods of analysis take place without change in
valence. Oxidation-reduction methods of analysis nearly always
involve a change in valence of the reacting substances. A study
of the principles underlying the latter type of reactions is an
essential preliminary if the methods based on them are to be
understood.
Theory.-The simplest type of oxidation-reduction reactions
are addition reactions. Thus, when oxygen gas unites with
hydrogen gas, forming water, O 2 + 2H2~2H20, the oxygen
is reduced and the hydrogen oxidized. When carbon burns in
the presence of oxygen to form carbon dioxide, C + 02~C02'
it is oxidized and the oxygen is reduced, and when carbon unites
with sulfur to form carbon disulfide, C
2S~CS2' the carbon
is oxidized and the sulfur reduced.
From the above simple illustrations, it is obvious that reduction
need not imply a reaction of hydrogen, since oxygen is reduced
by carbon, and that oxidation need not imply a reaction of
oxygen, since carbon is oxidized by sulfur. When one substance is oxidized, some other substance must be correspondingly
reduced, and conversely, when one substance is reduced, some
other substance must be correspondingly oxidized.
The oxidation-reduction reactions which take place in the
official assay processes are, for the most part, between electrolytes
in aqueous solution. Their quantitative value is based upon the
fact that metals, non-metals, "and their ions, under suitable
conditions, can be made to undergo a change in the quantity
of electric charge associated with them and that in the change
there exists a simple relationship between the quantity of
electricity lost or gained and the weight of the reacting substances. This relationship may be expressed according to Fara-
129
130
day's law as follows: A change of charge of one is equivalent to

the gain or loss of 96,500 coulombs of electricity for each formula
weight of element or group of elements involved. Since in every
oxidation-reduction reaction the charge lost or gained by one
substance must necessarily be gained or lost by another, it follows
that there is always a transfer of electricity in oxidation-reduct10n
reactions.
It has been shown by experiment that substances classified
chemically as reducing lose negative charges or electrons when
oxidized and that those classified as oxidizing gain electrons
when reduced. Oxidation, therefore, consists of the loss of
negative charge, and reduction consists of the gain of negative
charge. A unit negative charge is an electron according to the
electron theory of matter, and a unit positive charge, the proton,
is the charge left on an atom when it has lost an electron. Oxidation
may then be defined as the loss of one or more electrons by an atom
or ion, and reduction may be defined as the gain of one or more
electrons by an atom or ion.
When metallic iron is oxidized to the ferrous ion, each atom of
iron loses 2 electrons. If an electron is rep resEW ted by the negative sign (-), and the charge left on the atom by the positive
sign C+), the change may be indicated as F~Fe+t + 2(-).
Thus, when iron wire. is dissolved in hydrochloric -acid the
following reaction occurs:
(1) Fe
+ 2HC1~FeCb + H2
(2) Fe
Zero
+ 2H+ +
+ 2( +) +
or
2Cl-~Fe++
2( -) = 2( +)
+ 2Cl- + H2
+ 2( -) + zero.
Since Cl- is present on both sides of equation (2) the essential

2H+~Fe++ + H 2. During the reaction,
reaction is Fe
the iron has changed (been oxidized) from a neutral atom to an
ion bearing two positive charges through the loss of 2 electrons,
and the 2 hydrogen ions have each gained 1 electron (been
reduced to molecular hydrogen).
If the ferrous ion is further oxidized to the ferric state, it
loses another electron:
Fe++----> Fe+++
+ (-)
OXIDATION-REDUCTION METHODS
131
In the reaction
2Fe++
+ Ck-~2Fe+++ + 2Cl-
each ferrous ion loses 1 electron which is gained by a chlorine

atQm, for a negative charge on an ion indicates that it has gained
an extra electron. In the above case, the ferrous ion has been
oxidized, since it lost an electron, and chlorine has been reduced
to chloride ion, since it gained an electron.
When ferric chloride is reduced by stannous chloride, the
following reaction takes place: 2FeCla + SnCh~2FeCb +
SnC1 4 , which written ionically becomes
2Fe+++ + Sn++~2Fe++
2(55.84) 118.7
+ Sn++++
Each ferric ion gains one electron at the expense of the stannous
ions, each of which loses two electrons. The quantity of electricity gained by the iron is 96,500 coulombs for every 55.84 Gm. of
iron reduced, and that lost by the tin is 2 X 96,500 coulombs
for every 118.7 Gm. of tin oxidized. The chloride ion is in the
same state of oxidation before as after the reaction, since it
undergoes no change of charge.
In most cases, the change of charge which an atom, ion, or
radical undergoes in an uxidation-reduction reaction is numerically equal to the change in valence, i.e.:
1. When ferrous iron is oxidized to the ferric condition, the
charge associated with the iron atom changes from two to three:
2Fe++
+ 2H+~2Fe+++ + H2
and the valence also changes from two to three:

Cl
2Fe
/
'"
Cl
+ C1 2---2Fe-Cl
Cl
'0>
'"
Cl
2. When potassium permanganate is reduced, the manganese

atom gains five negative charges:
Mn7(+)~M!l++
+ 5(+)
132
and it changes from a valence of seven to a valence of two:
o
---+Mn
/""-/
(See assay of FeS04 page 136.)
""-/~
3. When potassium chlorate is reduced, however, thE). chlorine

atom changes from a charge of 5 ( +) to a charge of 1 ( - ), and
since the total change of polarity is the algebraic sum of the
charge carried by the atom on each side of the equation, the
change in polarity of the chlorine atom is six:
Clo<+)---+Clbut the change in valence experienced by the chlorine is four:
O-K
Cl=O
~
----+KCl
(See assay of KC10 s, page 149.)
Standard Solutions.-The standard solutions \Used in the

official assays by oxidation-reduction methods are: potassium
permanganate, potassium dichromate, iodine and bromine as
oxidizing agents, and oxalic acid and sodium thiosulphate as
reducing agents.
A normal solution of an oxidizing or reducing agent is one which
contains lin moles of reagent per 1,000 cc. where n represents the
change of charge which the reagent undergoes in the reaction.
The change of charge which oxidizing agents in the official
standard oxidizing solutions undergo is given below:
1. Permanganate ion-manganous ion
Mn04- + 8H+---+Mn++ + 4H 20
1( -) + 8( +)---+2 ( +) + 'zero + 5( +)
When the manganate ion is reduced to the manganous ion,
it gains five electrons or loses five positive charges. A normal
solution of potassium permanganate should therefore contain
U mole of KMn04 per 1,000 cc.
133
2. Dichromate ion~chromic ion

Cr 207 --+ 14H+~2Cr+++ + 7H 20
2(-) + 14(+)~6(+) + zero + 6(+)
Upon reduction to chromic ion, each dichromate ion loses
six positive c~arges. A normal solution of dichromate must
therefore contain U mole of K2Cr207 per 1,000 cc.
3. Iodine-iodide ion
12 21Zero
2( -) + 2( + )
Each atom of iodine gains 1 electron and supplies one positive
charge. A normal solution would therefore contain 1 mole of
iodide ion per 1,000 cc.
The change of charge which the reducing agents in the official
standard solutions undergo is as follows:
1. Oxalate ion~carbon dioxide

0 204-- - 200 2 + 2( - )
2( - )
zero + 2( - )
The change of charge is two per mole of oxalate ion, and a
normal solution of oxalic acid should contain U mole of oxalic
acid per 1,000 cc.
2. Thiosulphate
28 20 3- 4( -)
ion~tetrathionate
--
ion
8 40 6- 2( -) + 2( - )
The change of charge is one per mole of thiosulphate ion, and a

normal solution of sodium thiosulphate should contain one mole
of sodium thiosulphate per 1,000 CC.
Permanganate Methods.-Potassium permanganate volumetric solution can be standartlized easily, and it retains its concentration over long periods of time when proper precautions
are observed in its preparation and preservation. The reactions
of permanganate in solution are rapid. It also serves as an
indicator in titrations where it is used, since a very slight excess
of permanganate imparts to solutions a distinct pink color.
134
QUANTITATIVE PHARMACEUTICAL CH)EMISTRY

Exercise 37
Object.-To Prepare and Standardize 0.1 N Potassium

Permanganate.
Materials Required.-About 3.3 Gm. of potassium permanganate.
About 1 Gm. of reagent sodium oxalate.
Procedure.-l. "Dissolve about 3.3 Gm. of potassiul1t permanganate in
1000 cc. of distilled water in a flask and boil the solution for about fifteen
minutes. Stopper the flask and allow it to stand for at le!\'st two days before
filtering through asbestos."
Any organic matter which may be present in distilled water is

decomposed by potassium permanganate. Conlilequently, after
solution of the permanganate is effected, the liqu.id is allowed to
'stand for two days to insure the completion of the decomposition
reaction. The solution is then filtered through asbestos to
remove all traces of manganese dioxide which if present acts as a
catalyst to promote the formation of more manganese dioxide at
tile expense of permanganate ion.
2. "Ascertain its exact normality by titration again1;t reagent sodium
oxalate as follows:
"Weigh accurately about 0.2 Gm. of reagent sodium ()ialate, previously
dried, to constant weight at 1l0C., and dissolve it in 250 cc. qf distilled
water. Add 7 cc. of sulfuric acid, heat to about 70C. aI\d then Slowly add
the permanganate solution from a burette, with constant stirring, until a
pale pink color is produced which persists for fifteen seconds. The temperature at the conclusion of the titration should not be less than 6OC.
Calculate the normality of the solution and, if desired, adjust the solution
exactly to tenth-normal."
Titrate the mixture by adding the perman~anate solution

from a glass-stoppered burette, with continuous stirring of the
titration mixture until a drop of the permanganate causes a
permanent pink color. Read the top of the ITleniscus in the
burette instead of the lowest part. Using the first sample as a
preliminary run, titrate the two other portions of oxalate in the
same manner and base the calculations of normality upon them.
Sodium oxalate is the best standard to use in the standardization of potassium permanganate, since it can be obtained in
very pure condition. Sufficient sulfuric acid mu.st be .added to
keep the hydrogen ion concentration sensibly COIlstant throughout the titration, thereby preventing the formation of manganese
135
dioxide and supplying the hydrogen ions used up in the reduction

of the permanganate ion. Thus for every mole of permanganate
ion reduced, 8 moles of' hydrogen ion are required as shown in the
equation below. The top of the meniscus is read, since the
color of the permanganate solution renders impossible accurate
readings of the 10'Yest point.
The reactions involved are as follows:
5Na 2C 20 4 + 8H 2S0 4 + 2KMnOc----+5Na 2S04 + 10C0 2 + K 2S0 4
5(134.01)
2(158.03)
+ 2MnS04 + 8H 20.
5C 20 4-- 2MnOr + 16H+-)10C0 2 + 2Mn++
8H 20.
5 X 2( -) + 2( - )
16( +) = zero + 2 X 2( + )
lVIn04--)Mn++ giving up 5( +)
C204--~2C02 taking on 2( +)
Therefore 2 moles of Mn04- are equivalent to 5 moles of C 20 4--,

and 2KMn04 is equivalent to 5Na 2C 20 4.
The amount of sodium oxalate equivalent to 1,000 cc. of 0.1
N KMn04 calculated from the above equation is
~ ~ ;3~9190
= 6.7000 Gm. Therefore 1 cc. of 0.1 N KMn04 is equivalent

to 6.7000/1,000 = 0.006700 Gm. Na2C204. If a 0.25 Gm.
sample of sodium oxalate required 34.20 cc. of 0.1 N KMn04
for titration, the normality of the KMn04 solution would be
34.2
~'~~00670
X 0.1 = 0.10907, and the solution should be
labeled 0.10907 N KMn04.

When permanganate solutions are diluted to an exact normality, water freshly distilled from permanganate soiution should be
used to avoid the introduction of organic matter.
Potassium permanganate solution prepared as directed above
will retain its value for a long time if it is protected from sunlight
al].d from the entry of organic matter. It is best, however, to
restandardize the solution from time to time. Solution of
standard potassium permanganate withdrawn from the stock
bottle should never be returned to it. Solutions of oxidizing
agents should never be titrated from Mohr burettes, since the
oxidizing agent attacks the rubber and the solutions are thereby
weakened in concentration. The burette used to measure the
136
permanganate solution should be washed out immediately

after use to prevent the formation of a deposit of manganese
dioxide on the burette wall.
1. Define the following terms in accordance with the electron theory:
(a) oxidation, (b) reduction, (c) electron, (d) unit positive charge.
2. How much KMn04 is needed to prepare 500 cc. of 2N, 0.1200 N,
0.02 N, and 0.01 N KMn04 solutions, respectively?
3. Why is the titration of permanganate with a reducing agent carried
out in acid solution?
4. Why should Mohr burettes never be employed to measure permanganate solution?
DIRECT TITRATION METHODS
Exercise 38
To Determine the Purity of Ferrous Sulate.-Potassium

permanganate is especially well suited to the estimation of iron
and ferrous salts oxidimetrically. When the titration is carried
out in solution acidulated with sulfuric acid, the ferrous ion
is oxidized quantitatively to ferric ion.
Object.-Assay of Ferrous Sulfate.
Materials Required.-About 1 Gm. of ferrous sulfate.
25 cc. of diluted sulfuric acid.
0.1 N potassium permanganate.
Procedure.-"Dissolve about 1 Gm. of Ferrous Sulfate, accurately
weighed, in 25 cc. of diluted sulfuric acid, and titrate with, tenth-normal
potass~um permanganate until a permanent pink color is produced. Each
cc. of tenth-normal potassium permanganate corresponds to 0.01519 Gm.
of FeS04."
After dissolving the sample in 25 cc. of diluted sulfuric acid,

it is best to wash down the sides and neck of the flask with about
25 cc. of cold distilled water. The titration must be conducted
in acid solution to maintain a sensibly constant hydrogen ion
concentration as explained on page 134. The pink color of
permanganate is distinct when a drop in excess of that required
to oxidize all of the ferrous salt to the ferric state has been added.
The reaction is as follows:
10FeS04 + 2KMn04
10(151.9) 2(158.03)
+ 8H2S0,--+5Fe2(SO,)3 + 2MnSO,
.
137
OXIDATION.REDUCTION METHODS
or, written ionic ally,

10Fe++
+ 2MnOc + 16H+-)10Fe+++ + 2Mn++ + 8H 0

2
In the above equations the water of crystallization is omitted,

since the U.S.P. assay determines the content of FeS04.
316.06 Gm. Qf KMn04 is equivalent to 1,519 Gm. of FeS04
and each cubic centimeter of 0.1 N KMn04 is equivalent to
1,519
100 X 1,000 = 0.01519 Gm. FeS04. The 100 in the denominator
is obtained by taking the change of charge of 2KMn04 = 10
multiplied by 10, the dilution of a 0.1 N solution.
If the per cent of iron in the sulfate is desired, it can be calculated by substituting its equivalent, 55.84, for that of FeS04.
'-Thus each cubic centimeter of 0.1 N KMn04 is equivalent to
10 X 55.84
100 X 1,000 = 0.005584 Gm. Fe. By the same procedure, the
equivalent of FeS04.7H 20 can be calculated.
Calculate the per cent Fe, FeS04, and FeS04.7H 20 equivalent
to the sample assayed. The U.S.P. requires that ferrous sulfate
contain 55.36 to 57.07 per cent FeS04. Compare your results
with the official requirement.
1. Why is it unnecessary to employ an indicator in direct titrations with

KMnO,?
2. What quantity of electricity undergoes transfer in the oxidation of
two moles bf FeSO, to Fe2(SO.).?
. 3. Does the change of charge on the iron in question 2 correspond with
the change in valence?
4. What must be the normality of a solution of KMnO. in order that each
cubic centimeter shall be equivalent to 0.1 per cent FeSO.?
5. How much FeSO. would be required to prepare 500 ce. of 0.1 N solution
of FeSO . 7H 20?
6. What other official ferrous salts are assayed by methods analogous to
that given above?
Exercise 39
To Determine the Purity of Reduced Iron.-In this assay the

metallic iron is converted quaptitatively into ferrous ion and
estimated oxidimetrically by titration with potassium permanganate solution.
138
Object.-Assay of Reduced Iron.

Materials Required.-About 1 Gm. of reduced iron.
10 Gm. of mercuric chloride.
Procedure.-1. "Triturate thoroughly about 1 Gm. of Reduced Iron,
weigh accurately, place it in a 100 cc. volumetric flask, and add 10 Gm. of
finely powdered corrosive mercuric chloride and 50 cc. of boiling distilled
water."
The reduced iron is triturated to produce a uniform mixture

in order that a representative sample may be obtained for assay.
Since reduced iron always contains some oxide, the metallic
iron in the sample is oxidized by boiling it in solution with mercuric chloride. The mercuric chloride oxidizes the metallic
iron to the ferrous state without changing any oxide present.
HgCh
+ Fe~FeCh + Hg
or, written ionically,

Hg++
+ Fe~Fe++ + Hg
2. "Heat the mixture for ten minutes on a water oath, Rhaking it frequently, then- fill the flask to the 100 cc. mark with distilled water, recently
boiled and cooled, and cool the mixture to room temperaturt Again fill
the flask to the mark with distilled water, stopper, agitate the contents well,
and allow to stand for a few minutes."
The solution is diluted to exactly 100 cc. so that an aliquot.

portion representing one-fifth of the weight of sample (20 cc.
of solution) may be obtained.
3. "Now filter the contents of the flask, and immediately titrate 20 cc.
of the filtrate to which has been added 20 cc. of diluted sulfuric acid, with
tenth-normal potassium permanganate, until a permanent pink color is
produced. Each cc. of tenth-normal potassium permanganate is equivalent
to 0.005584 Gm. of Fe."
The aliquot portion is filtered to free it of any finely divided

mercury or iron oxide which may be suspended in the solution.
The titration is conducted in acidulated solution in a manner
similar to that described in the preceding assay.
The reactions involved in the titration and the calculations
are also similar. Write all reactions and calculate the purity
of the sample assayed.
139
1. Does the iron gain <lr lose electrons when oxidized to the ferrous state
by HgCI.? What quantity of electricity does it gain or lose per mole of Fe?
2. Given pure iron, how might a solution of potassium permanganate be
standardized against it?
3. If the iron in.a 0.15 Gm. sample of iron ore requires 15.0 ce. of 0.1 N
KMnO. for oxidation from Fe++ to Fe+++, what is the percentage of Fe in
the ore?
Exercise 40
Object.-Assay of Solution of Hydrogen Peroxide.

Materials Required.-About 2 cc. of solution of hydrogen peroxide.
Procedure.-" Measure accurately 2 cc. of Solution of Hydrogen Peroxide
and transfer it to a suitable flask containing 20 cc. of distilled water. Add
20 cc. of diluted sulfuric acid, and titrate with tenth-normal potassium
permanganate. Each cubic centimeter of tenth-normal potassium permanganate is equivalent to 0.001701 Gm. of H 20 2."
The reasons for the various steps in the above assay have
already been given under the assay of ferrous sulfate. This
assay illustrates a case where an oxidizing agent, H 20 2, serves
as a reducing agent for KMn04 and is itself reduced. When
H 20 2 acts as a reducing agCent, 1 Gm.-atom of oxygen from each
mole of H 20 2 is capable of uniting with 1 Gm.-atom of oxygen
from the substance reduced (KMn04.) to form 1 mole of oxygen
gas:
2HO
+0
(from oxidizing agent)--+H 20
+O
In the assay reaction 5 Gm.-atoms of oxygen from 5 moles of

H 20 2 unite with 5 Gm.-atoms of oxygen from 2 moles KMn04
to form 5 moles of oxygen gas:
5H 20 2 4- 2KMn04
5(34.02) 2(158.03)
+ 3H2S04~K2S04 + 2MnS04 + 8H 20
+ 50
Each cubic centimeter of 0.1 N KMn04 is equivalent to

5 X 34.02
100 X 1,000 = 0.001701 Gm. H 20 2
140
To comply with the U.S.P. definition, solution of hydrogen

peroxide should contain not less than 2.5 and not more than 3.5
per cent of H 20 2 Calculate the percentage of H 20 2 in the sample
assayed and compare the strength of the solution with the U.S.P.
requirement.
1. A 2 cc. sample of hydrogen peroxide required 8.5 cc. of KMnO. solution
for titration. If each cubic centimeter of KMnO. is known to be equivalent
to 0.007294 Gm. Fe, what percentage of H 20 2 does the sample contain?
2. Is H 2SO. necessary to supply a concentration of H+ to combine with
the oxygen liberated from the perm~nganate to form water? Why is the
H 2SO. necessary?
3. Sodium perborate when dissolved in diluted sulfuric acid solution yields
NaHSO. + H 3BO i + H 20 2. Ascertain the method of assay of sodium
perborate, balance the reaction involved in the assay, and compare the
method with that employed in the assay of hydrogen peroxide.
4. Could lead dixoide be assayed in this way? (See U.S.P. reagent
Pb0 2.)
TABLE XIV.-OFFICIAL SUBSTANCES ASSAYED BY DIRECT TITRATION WITH
0.1 N KMnO.
Substance
Amount
used,
Gm. or
ee.
U.S.P.
Ferrous sulfate ..........
Hydrogen peroxide solution ..................
Hydroxylamine hydrochloride, R ...........
Indigo carmine, R, '.' ....
Iron, reduced ...........
Lead peroxide, R ........
Sodium perborate ........
Sodium peroxide, R ......
Equivalent of
1 cc.
0.1 N
KMn04
Official tequirement,
per cent
0.01519 FeSO. = 54.36 to 57.07
0.001701 H 20 2 = 2.5 to 3.5
0.1
0.3
1
0.5
0.25
0.7
0.003475
0.01333
0.005584
0.01196
0.0008
0.0039
NH 20H.HCI = 95
C 16 H,02N 2(S03Na).
Fe = 90
Pb0 2 = 90
Available O 2 = 9
Na20, = 90
.
= 82
R. = reagent.
Potassium permanganate is the only official substance assayed

by direct titration with 0.1 N oxalic acid. The principle involved
141
is the same as that employed in the standardization of potassium

permanganate against sodium oxalate (see page 134).
INDIRECT TITRATION METHODS
Exercise 41
Object.-Assay of Calcium Gluconate.

Materials Required.--Q.5 Gm. of calcium gluconate.
Ammonium oxalate, T.S. (3.5 Gm. ammonium oxalate in 100 cc. of water).
Dilute sulfuric acid (1 in 3).
Procedure.-l. "Weigh accurately about 0.5 Gm. of Calcium Gluconate,
dried to constant weight in a desiccator over sulfuric acid, place it in a
25O-cc. beaker, add 2 cc. of hydrochloric acid and 100 cc. of distilled water.
Heat the mixture to boiling, make the solution alkaline with ammonia T.S.
and add, with stirring, an excess of hot ammonium oxalate T.S. Heat the
mixture on a water bath during one hour, filter through hardened filter paper
and wash thoroughly with warm distilled water."
The calcium gluconate reacts with the HCI as follows:
OH 20H(OHOH)4.000 20a.H 20
+ HOI
2CH 20H(OHOH)4_
OOOH + OaOl2 + H 20
448.27
When ammonium oxalate is added to the reaction mixture, the
calcium ion is quantitatively precipitated as oxalate and the
above reaction is forced to the right,
OaOl2 + (NH4)20204~Oa0204 + 2NH40l

The precipitation is carried out in hot solution and the mixture
is digested to secure a coarse grained precipitate.
2. "Puncture the filter paper, wash the precipitate into a beaker by
means of a stream of hot distilled water, followed oy 30 cc. 'Of dilute (1 in 3)
sulfuric acid. Heat the solution, if necessary, to 60C. and titrate with
tenth-normal potassium permanganate. Each cubic centimeter of tenthnormal potassium permanganate i~ equivalent to 0.002804 Gm. of CaO."
The precipitate .is washed with warm water to remove the

excess ammonium oxalate. Upon the addition of diluted sulfuric
acid, the calcium is precipitated as sulfate leaving free oxalic
acid in solution,
Oa0204
+ H2S04~CaS04 + (OOOH)2.
142
and the amount of oxalic acid liberated is found by titration with

standard KMn04 solution. The reaction indicated proceeds to
completion because the oxalic acid formed is consumed by titration with the permanganate.
This is an indirect method since neither the calcium nor the
gluconic acid is determined directly but the calcium is determined by the indirect process of estimating the amount of oxalic acid
with which it combined.
Calculate the per cent of calcium gluconate in the sample
assayed.
1. Calcium gluconate ilS required to yield not less than 12.4 and not more
than 12.8 per cent of CaO. Within what limits of per cent purity would
calcium gluconate conform to these requirements?
2. What amounts of anhydrous calcium lactate might 5 grain tablets of
calcium lactate contain and meet the requirements of the N. F.?
TABLE
XV.-OFFICIAL
SUBSTANCES
ASSAYED
BY
PRECIPITATION
WITH
AMMONIUM OXALATE AND TITRATION OF THE OXALIC ACID FROM

THE PRECIPITATE WITH
0.1 N
KMnO,
Amount Equivaused,
lent of
Substance
Gm. or
ce.
1 ce.
0.1 N
KMnO.
----------- --- --- --U.S.P.

Calcium bromide ............ .
Calcium gluconate ... ....... .
Calcium lactate ............. .
N.F.
Calci urn carbonate, tablets of.
Calcium gluconate, ampuls of.
Calcium lactate, tablets of ....
Official
requitement,
per cent
----------
0.4
0.5
0.5
0.009996 CaBr, = 84 to 94
0.002804 C"O = 12.4 to 12.8
0.01091 (CH,CH.OH.COO),Ca = 98
0.6"
0.5a
1.5
0.005004 CaCO, = 92.5 to 107.5'

0.02242 C"H,,014Ca.H,O = 95 to lOS'
0.01541 (CH,CH.OH.COO),Ca.5H,0 = 92.5
to 107.5'
Weight of ingredients sought.

, Per cent of the labeled amount.
The residual titrations in the official assays with potassium

permanganate solution are of three types, namely: (1) An excess
of standard potassium permanganate solution is employed to
oxidize a substance, and the amount in excess is determined by
143
residual titration with standard oxalic acid solution; (2) an excess

of standard oxalic acid solution is added to a substance and the
excess of oxalic acld is titrated with standard permanganate
sulution; (3) an excess of ferrous sulfate solution is added to an
oxidizing agent, and the amount of ferrous sulfate in excess is
titrated with st:;),ndard permanganate solution.
The following exercise is an example of a residual titration
method in which an excess of standard permanganate solution
is employed to oxidize a substance, and the amount in excess is
determined by residual titration of standard oxalic acid solution.
Exercise 42
Object.-To Prepare and Standardize 0.1 N Oxalic Acid.

Materials Required.-6.45 Gm. of reagent oxalic acid.
Procedure.-Dissolve 6.45 Gm. of reagent oxalic acid in sufficient distilled water to measure 1,000 cc. Ascertain its exact strength by titration
against freshly standardized 0.1 N potassium permanganate and adjust to
exacu normality or to a known strength of approximate normality.
The purest oxalic acid obtainable varies in composition due to

differences in its moisture content. If the acid is heated sufficiently to drive off absorbed moisture, it tends to lose water of
crystallization. Consequently, a solution of approximate concentration is prepared, and itll normality is determined by
titration against a standard solution, in this case, 0.1 N KMn04.
The titration is conducted as follows:
Measure accurately from a burette about 30 cc. of the acid
solution into a beaker and dilute with about 200 cc. of distilled
water. Add 7 cc. of sulfuric acid, heat to about 70C. and then
slowly add the standard permanganate solution from a glassstoppered burette, with constant stirring, until a p-ale pink color
is produced which persists for 15 sec. The temperature at the
conclusion of the titration should not be less than 60C. Calculate the normality of the sOlution and, if desired, adjust the
solution to exactly 0.1 N. The calculations and reactions
are similar to those described in the preparation of 0.1 N KMn04.
The probability of error is minimized by standardizing the
oxalic acid solution against the same permanganate solution
with which it is to be used in analyses. The normality of an
144
oxalic acid solution may be determined by titration with standard

alkali as follows:
Dilute 25 cc. of 0.1 N NaOH accurately measured from a
burette with 25 cc. of distilled water, add 2 to 4 drops of phenolphthalein T.S. as indicator, and heat the solution ~o boiling.
Titrate this solution with 0.1 N oxalic acid until the pink color
disappears. The solution should be brought to a gentle boil at
the end to expel all CO 2 which, if present, would affect the
phenolphthalein indicator.
The oxalic acid is dibasic; that is, one molecule requires two
of sodium hydroxide for neutralization. Since 1 cc. of 0.1 N
acid is equivalent to 1 cc. of 0.1 N alkali, the normality of this
solution may be calculated easily:
Cc. alkali X N
Cc. am'd
= normal'lty 0 f am'd
, Oxalic acid solution deteriorates on standing and must be

restandardized from time to time.
Questions and Problell}s
1. Why is hot titration employed?
2. What is the eq~ivalent weight of oxalic acid as an\ acid; as a reducing
agent?
3. Write the reaction involved in the standardization of oxalic acid
against sodium hydroxide solution.
4. How much oxalic acid is required to prepare 1,000 cc. of solutions of the,
following respective normalities: (a) 0.5000, (b) 0.2500, (c) 0.200?
Exercise 43
Object.-Assay of Sodium Nitrite.

Materials Required.-1 Gm. of sodium nitrite.
50 cc. of 0.1 N potassium permanganate.
5 cc. of sulfuric acid.
0.1 N oxalic acid.
Procedure.-l. "Dry about 1 Gm. of Sodium Nitrite to constarit weight
over sulfuric acid, weigh accurately in a stoppered weighing-bottle, dissolve the salt in a volumetric flask with sufficient distilled water to make
100 cc., and add 10 cc. of this solution, from a pipette, to a mixture of 50 cc.
of tenth-normal potassium permanganate, 100 cc. of distilled water, and 5
cc. of sulfuric acid. When adding the sodium nitrite solution, immerse the
tip of the pipette beneath the surface of the permanganate mixture."
145
Sodium nitrite is deliquescent and is therefore directed to be

dried to constant weight in a desiccator over sulfuric acid
previous to weighing. Because of this property, the sample is
also directed to be weighed in a stoppered weighing bottle.
Nitrous acid is oxidized by the permanganate to nitric acid.
If the solution of sodium nitrite were acidulated with sulfuric
acid, nitrous acid, which is volatile, would be lost. The sodium
nitrite is therefore introduced beneath the surface of the acidulated permanganate solution.
2. "Warm the liquid to 40C., allow it to stand for five minutes, and add
25 cc. of tenth-normal oxalic acid. Heat the mixture to about 80C., and
titrate with tenth-normal potassium permanganate. Each cubic centimeter
of tenth-normal potassium permanganate is equivalent to 0.003450 Gm. of
NaN0 2. "
The oxidation of nitrous to nitric acid proceeds slowly at

ordinary temperature, but it is rapid at 40C. The warm mixture
is allowed to stand for 5 min. to insure completion of the oxidation. The excess 0.1 N oxalic acid is then determined by titration with permanganate until the pink permanganate color is
obtained.
The reaction may be represented as taking place in two stages,
viz.:
5HN0 2
2NaNO. + H.SOc~Na2S04 + 2HNO.

2(69.01)
2KMn04 + 3H2S04~5HN03 + K 2S04
+ 2MnS04
+ 3H 20
In this reaction each nitrogen atom loses two electrons and

undergoes a change of polarity of two. Therefore, 1 cc. of 0.1
N KMn04 is equivalent to 2 X
N aN0 2
1~9.~11,000 = 0.003450
Gm.
Calculate the purity of the sample assayed.

1. Why should the tip of the pipette be immersed beneath the surface 0:
the permanganate solution when the solution of sodium nitrite is added?
2. Why is the reaction mixture warmed and allowed to stand?
3. Write ionic ally the reactions involved in the above assay.
4. How much 0.0985 N KMnO. would be required to titrate 0.8544 Gm.
of NaNO. if the NaNO. is 92 per cent pure?
6. Could nitrous acid and other nitrites be assayed by the,above method?
146
Sodium nitrite and potassium nitrite, reagent, are the only

official substances assayed by this method.
The following exercise illustrates residual titration methods in
which an excess of standard oxalic acid solution is employed to
precipitate a substance quantitatively with subsequent residual
titration of the excess oxalic acid by standard permanganate
solution.
Exercise 44
To Determine the Purity of Calcium Carbonate.-Calcium
and lead salts can be assayed volumetrically by precipitation
with standard oxalic acid added in excess and subsequent titration of the excess acid with standard potassium permanganate
solution.
Object.-Assay of Precipitated Calcium Carbonate.
Materials Required.-About 0.4 Gm. of precipitated calcium carbonate.
10 ce. of diluted hydrochloric acid.
100 cc. of 0.1 N oxalic acid.
Ammonia water, 10 per cent.
Diluted sulfuric acid.
Procedure.-l, "Dry about 0.4 Gm. of Precipitated Cal~um Carbonate
to constant weight at 200C., weigh accurately, dissolve it in a mixture of
10 cc. of diluted hydrochloric acid and 10 cc. of distilled water, and boil the
solution to expel all carbon dioxide."
Calcium carbonate is soluble in hydrochlorjc acid because the

product of the concentration of calcium and carbonate ions
becomes less than the solubility product of calcium carbonate.
When a strong acid is added to calcium carbonate, the concentration of CO a-- ion is decreased, due to the formation of slightly
ionized carbonic acid:
CaCO a
Ca++
C0 3- - + 2H+
+ CO a-H 2CO a
CO 2
+ HiO
The increase in H+ concentration 'displaces the equilibria

toward the right and leads to solution of the calcium carbonate
in hydrochloric acid. The CO 2 formed in the reaction is expelled
by boiling.
147
2. "Transfer this solution to a 200 cc. volumetric flask, add 100 cc. of
tenth-normal oxalic acid, render it alkaline with ammonia T.S., shake the
mixture well, and allow it to stand for three hours at from 60 to 70C., or
over night at room temperature. Cool the liquid if necessary, fill the flask
with distilled water to 200 cc., mix well, filter through a dry filter into a
dry.flask, and reject the first 20 cc. of the filtrate."
The 0.1 N oxalic acid is added to the solution contained in the

200 cc. volumetric flask and the whole is made alkaline with
ammonia T.S. added slowly with continuous stirring. Ammonium oxalate is formed, and it reacts with the calcium chloride
to form ammonium chloride and insoluble calcium oxalate. The
excess of ammonium oxalate present decreases the solubility of
calcium oxalate to a negligible quantity by common ion effect.
The solution is digested at 60 to 70C. to promote the growth of
larger crystals and to permit complete precipitation of the
calcium oxalate. The solution is cooled, since calcium oxalate is
more soluble in hot than in cold water, its solubility being 5.6
mg. per liter at 20C. and 14 mg. at 95C. The solution is
made up to a precise volume, 200 cc., and an aliquot portion,
100 cc., is taken for residual titration. This procedure eliminates
the tedious washing which would be required to render the crystalline calcium oxalate free of ammonium oxalate. The filtration of the aliquot portion is best conducted by collecting the first
20 cc. of filtrate, which is to.be rejected, in a cylindrical graduate,
then transferring the funnel to a 100 cc. volumetric flask and
filtering an additional quantity of solution sufficient to fill it
exactly to the graduation mark.
3. "Acidulate 100 cc. of the filtrate (corresponding to one-half of the
Precipitated Calcium Carbonate taken) with diluted sulfuric acid, then
add 25 cc. more of the diluted sulfuric acid, warm the solution to about
60C., and titrate with tenth-normal potassium permanganate. Each
cubic centimeter of tenth-normal oxalic acid is equivalent to 0.005004 Gm. of
CaCO,."
The filtrate is transferred quantitatively to a 250 cc. Erlenmeyer flask, acidulated with sulfuric acid and titrated with 0.1
N KMn04 to determine the excess oxalic acid.
The reactions involved are:
1. Solution of CaCO a in HCI:
CaCO a + 2HCI-4CaCI 2
CO 2
+H
148
2. Addition of NH 40H to H 2C 20 4:
H 2C20 4 + 2NH40H~(NH4)2C204
+ 2H 20
3. Double decomposition between CaCl 2 and (NH4)2C 20 4:

CaCl 2 +
(NH4)2C204~CaC204
+ 2NH 4CI
4. Filtrate of (NH 4)2C 20 4 acidulated with H 2S0 4:

(NH4)2C20 4
+ H2S04~H2C204 + (NH4)2S0<t.
5. Residual titration of excess H 2C 20 4 with KMn04:

5H 2C20 4 + 2KMn04
+ 3H2S04~K2S04 + 2MnS04
+ lOC02 + 8H 0
2
The calculation of the purity of the sample may be made

according to the following:
One cubic centimeter of 0.1 N oxalic acid is equivalent to
100.08
2 X 10 X 1,000 = 0.005004 Gm. CaCO s
[(N X cc. H 2 C 204) - (N X cc. KMn04 X 2)LX 0.005004
Wt. of sample
X 100
= Plir cent purity
Note that the number of cubic centimeters of KMn04 solution

is multiplied by two, since only one-half of the exce~s oxalic acid
is represented in the aliquot portion used in the residual titration.
1. Why is the solution rendered alkaline with NH 40H?
2. How does common ion effect serve to make the precipitation of calcium
oxalate complete?
8. Why is the mixture digested at 60 to 70C.?
4. Why is the solution cooled previous to dilution and filtration?
5. Name several official calcium compounds which are assayed by methods
analogous to that given above.
6. Look up the official method of assay for solution of lead subacetate.
Explain each step in the procedure and write all reactions.
7. An unknown calcium salt weighing 0.25 Gm. assayed by the above
method was treated with 100 cc. of 0.1010 N oxalic acid. Upon residual
titration of an aliquot portion representing one-half of the excess oxalic
acid, 22.45 cc. of 0.0982 N KMn04 was required. To what percentage of
Ca, CaO, CaCO" and CaCh, respectively, is the calcium salt-equivalent?
TABLE
149
XVI.-OFFICIAL SUBSTANCES ASSAYED BY PRECIPITATION WITH
EXCESS STANDARD OxALIC ACID SOLUTION AND RESIDUAL TITRATION

OF THE EXCESS OXALIC ACID WITH
Amount
used,
Gm.
or cc.
Substance
U.S.P.
Calcium carbonate, precipitated
Calcium chloride ..............
Calcium chloride, fused, R .. ...
Calcium hydroxide .............
Chalk, prepared ...............
Lead acetate ............. .... .
Lead monoxide, R .............
Lime, R ......... ,."."., .. "
N.F.
Calcium chloride, ampuls of .....
0.4
0.3
0.3
0.3
3.5
5
0.4
1
O,25 a
Lead monoxide ... , .... , ....... 0,4

Lead subacetate, solution ......
1
Lead subacetate, solution, dilute 50
Lime (calcium oxide) ..... , ., ...
R.
0.1 N
Equivalent of 1
cc. 0.1 N
H 2C 2O.
KMnO.
Official
requirement,
per cent
CaCO. = 98
CaCh = 75 to 85
CaCl. = 94
Ca(OHh = 95
CaCO. = 97
Pb(C.H 3 0.). =
85.31 to 89.57
<;>.0116 PbO = 98
0.002804 CaO = 95
0.005004
0.00555
0.00555
0.003705
0.005004
0.01626
0.007351 CaCI..2H 20 = 95
to 105'
0.01116 PbO = 97
0,01036 Pb = 22.5
0.01036 Pb = 0.7 to
O.8W/V
0.002804 CaO = 95
reagent.
Per cent of the labeled amount.
Potassium permanganate, U.S.P., and manganese dioxide and

tablets of potassium permanganate of the N.F. are assayed by
treating the sample with an excess of standard oxalic acid and
titrating the excess of acid with standard permanganate.
The following assay illustrates a method involving reduction of
an oxidizing agent by means of an excess of ferrous sulfate, and
residual titration of the excess ferro~s sulfate (unoxidized)
by standard permanganate solution;
Exercise 45
Object.-Assay of Potassium Chlorate.

Materials Required.--O.1 Gm. of potassium chlorate.
150
100 cc. of acid ferrous sulfate solution prepared by dissolving 7 Gm. of

ferrous sulfate crystals (FeSO . 7H20) in 90 cc. of freshly boiled distilled
water and adding sufficient sulfuric acid to make 100 cc .
20 cc. of manganous sulfate solution prepared by dissolving 11 Gm. of
manganese sulfate (MnSO .4H20) in 50 cc. of distilled water and adding
sufficient sulfuric acid to make 100 cc.
Procedure.-l. "Weigh accurately about 0.1 Gm. of Potassium Chlorate,
dissolve it in 10 cc. of distilled water in a 250-cc. flask, and add, from a
burette, 35 cc. of acid ferrous sulfate T.S. Prepare a valve-stopper by taking a piece of rubber tubing of convenient diameter and about 5 cm. in
length, place a piece of glass rod in one end, and slip the other end over a
glass tube which passes through a perforated stopper of a size to fit the
flask used. Cut a longitudinal slit about 15 mm. long in one side of the rubber tube about midway of its length. Insert this stopper in the flask and
boil the mixture for ten minutes."
When potassium chlorate is treated with ferrous sulfate T.S.

in acid solution, the ferrous salt is oxidized to the ferric state
and the potassium chlorate is reduced to potassium chlqride:
KCI0 3 + 6FeS04
122.55
CI0 3-
+ 3H2S0c-~KCI + 3Fe2(S04)3 + 3H 20
+ 6Fe++ + 6H+
or
= CI-
+ 6Fe+++
Sufficient acid must be present to supply an abhndance of H+

to satisfy the equation and to prevent the hydrolysis of the ferric
sulfate. If oxygen from the air were allowed to 'enter, it would
oxidize some of the FeS04 to Fe2(S04)3. The reaction is, therefore, carried out in a flask fitted with a Bunsen valve which allows
steam to escape but prevents the entrance of air.
2. "Cool the mixture, add 10 cc. of manganous sulfate T.S., and titrate
the excess of ferrous sulfate with tenth-normal potassium permanganate."
The mixture should be cooled in the flask protected from air

and the manganous sulfate solution added just before titration
of the residual ferrous sulfate with standard potassium pe,rmanganate solution. The presence of manganous sulfate .is
essential in the reaction mixture. The manner in which it
functions is not clearly understood, but it is known that in the
presence of chlorides more permanganate is used up than is
necessary to oxidize the ferrous salt to the ferric condition. The
151
manganous ions react with permanganate ions to form manganese ions Mn++++. In this state of oxidation, the manganese
is unstable in acid solution, and it is reduced more readily by
ferrous ions than by chloride ions.
3. "Conduct a blank experiment with another portion of 35 cc. of acid
ferrous sulphate. T.S., measured from a burette, and subtract the result
of the former titration from that of the latter. Each cubic centimeter of
tenth-normal potassium permanganate is equivalent to 0.002043 Gm. of
KClO . "
A blank experiment is conducted employing all of the reagents

used in the assay bu<without the addition of potassium chlorate.
This procedure is necessary, since the exact concentration of the
ferrous sulfate T.S. is not known. The difference between the
volume of 0.1 N KMn04 consumed in the blank experiment
and in the assay gives the number of cubic Ilentimeters of 0.1
N KMn04 equivalent to the sample of potassium chlorate.
Each cubic' centimeter of 0.1 N KMn04 consumed in the
titration of the residual ferrous sulfate is equivalent to
122.55
6 X 10 X 1,000 = 0.002043 Gm. KCIO a
If 50 cc. of ferrous sulfate T.S. requires 49.5 cc. of 0.1 N
KMn04 in the blank experiment and the residual ferrous sulfate
in the assay requires 4.5 cc. of 0.1 N KMn04, the 0.1 Gm. sample
of Rcio a is equivalent to 49.5 - 4.5 = 45 cc. of 0.1 N KMn04.
The"per cent purity of the KCIO a may then be calculated:
O.OQ2~~~ X 45 X 100 = 91.93 per cent

1. Why is the reaction carried out in a flask fitted with a Bunsen valve?
2. Explain how the equivalent weight of KClO. is derived on the basis of
change of charge.
'"
3. Why is it necessary to run a blank experiment?
Potassium chlorate and tablets of potassium chlorate are the

only official substances assayed by this method.
Dichromate Methods.-In certain official preparations containing ferrous salts the percentage of iron salt cannot be accu-
152
rately estimated by oxidation in acidulated permanganate

solution owing to the presence of sugars which reduce this reagent. Potassium dichromate possesses two distinct advantages
over potassium permanganate as a volumetric reagent, namely, it
can be obtained very pure, and it does not deteriorate upon aging
if properly kept.
Determination of the End Point.-The end point is determined
in the officiaJ assays by the use of diphenylamine as the indicator.
In the presence of sugars, the violet color formed by this indicator
is obscured somewhat by the':ormation of the green chromic ion.
The end point of titration may also be determined by the use
of an "outside indicator" as follows: The end point of titration
is found by placing a drop or two of freshly prepared potassium
ferricyanide solution in a number of the depressions in a porcelain
spot plate. A drop of the solution being titrated is brought in
contact with this indicator by means of a stirring rod at frequent
intervals in the titration. As long as ferrous iron is present, a
blue coloration results when the solution comes in contact with
the indicator, the end point being reached when the blue color
is no longer produced. It is usually advisable 0 run a preliminary titration so that it will not be necessary to withdraw an
undue quantity of reaction solution during the titratiqn.
The following reaction takes place between ferrous sulfate
and potassium ferricyanide:
3FeS04
+ 2KaFe(CN)6--Fea[Fe(CN)6]2 + 3K S04
2
blue
Exercise 46
Object.-Preparation of 0.1 N Potassium Dichromate.

Materials Required.-1.7 Gm. of pure potassium dichromate.
Procedure.-Pulverize about 1.2 Gm. of pure potassium dichromate and
dry the product to constant weight at 120C. Accurately weigh exactly
0.9807 Gm. of dried K 2Cr 207 and dissolve it in sufficient distilled water to
make 200 cc. at the temperature at which the volumetric flask has been
calibrated. If the dichromate used is pure, the solution will be exactly
tenth-normal.
Each chromium atom in K 2Cr 207 has a polarity of 6( +) and

upon reduction two trivalent chromic ions are formed. Each
153
chromium atom undergoes a change in polarity of three. Since

two atoms of chromium undergo the change, the total change in
polarity is 2 X 3 = 6. ' A normal solution of K 2 Cr 207 should,
therefore, contain one-sixth of a mole per 1,000 cc.
An approximately 0.1 N solution of K 2Cr 207 can be standardized against pure iron wire or 0.1 N sodium thiosulfate solution
(see page 158).
Exercise 47
Object.-Assay of Mass of Ferrous Carbonate.

Materials Required.-About 1 Gm. of mass of ferrous carbonate.
15 .cc. of diluted sulfuric acid.
0.1 N potassium dichromate.
Diphenylamine, T.S. (1.0 Gm. of diphenylamine in 100 cc. of reagent
H 2S0 4 ).
Procedure.-" Dissolve about 1 Gm. of Mass of Ferrous Carbonate,
accurately weighed, in 15 cc. of diluted sulfuric acid, dilute the solution with
distille'd water to about 150 cc., and immediately titrate with tenth-normal
potassium dichromate, using about 0.5 cc. of diphenylamine T.S. as the
indicator. Each cubic centimeter of tenth-normal potassium dichromate is
equivalent to 0.01158 Gm. of FeC0 3 ."
The mass of ferrous carbonate dissolves in diluted sulfuric

acid to form ferrous sulfate, viz.:
Sufficient acid in excess of that required for the formation of

ferrous sulfate is needed to supply a constant source of hydrogen
ion:
The solution is diluted to facilitate the reaction in the presence

of the suspended matter and tQ minimize the error caus~d by
the forma.tion of the green chromic ion which tends to obscure the
violet color of the diphenylamine at the end point. The titration
should be conducted as soon as possible after solutlon of the
sample has been effected to avoid as much as possible the oxidation of ferrous to ferric iron by the action of atmospheric oxygen.
154
The reactions which occur in the assay are:

FeCO a + H 2SOc-?FeS04 + H 20 + CO2
115.84
6FeS04 + K2Cr207 + 7H2S04~3Fe2(S04)a + K 2S04
+ Cr2(S04)a + 7H 20
or
6Fe++
+ Cr 02-- + 14H+~6Fe+++ + 2Cr+++ + 7H 20

2
Each Fe++ undergoes a change of charge of one. Each cubic

centimeter of 0.1 N K 2Cr 207 consumed in the titration is thereG m. F e CO a C a 1. I
fore eqmva
ent t 0 10115.84
X 1,000 = 001159
.
culate the per cent of iron and the per cent of ferrous carbonate
in the sample assayed. The U.S.P. requires that mass of ferrous
carbonate contain 36 to 41 per cent of FeCO a. Compare the
per cent of ferrous carbonate found in the assayed sample with
the official requirement.
1. Why must the solution be acid in reaction during titration?
2. If three pills of ferrous carbonate of equal weight employed as a sample
require 16.25 cc. of 0.1 N K 2Cr 207, what amount of Fe andlof FeCO"
respectively, does each pill contain?
TABLE XVII.-OFFICIAL SUBSTANCES ASSAYED BY DIRECT TITRATION WITH
0.1 N K 2Cr 207
Amount
used
Equivalent of
1 cc.
Official
requirement,
per cent
U.S.P.
Mass of ferrous carbonate .........
1 Gm.
0.01158
Pill of ferrous carbonate ..........
3 pills
0.01158
FeCO, = 36 to
41
FeCO, = 0.06
Gm. per pill
2Gm.
0.01158
Substance
N.F.
Saccharated ferrous carbonate ......
FeCO, = 15
CHAPTER IX
OXIDATION AND REDUCTION.
METHODS
IODOMETRIC
Iodometry is the process of determining iodine volumetrically,

and assay methods which are based upon the quantitative determination of specific quantities of iodine are termed iodometric
methods. In these methods, the reversible reaction 12
21is applied, since the reaction from left to right 12----)021- can be
made to go to completion in-the quantitative oxidation of thiosulfate, arsenite, and other reducing agents, and the reaction from
right to left 12~2I- can be made to go to completion in the
quantitative reduction of permanganate, dichromate, arsenate,
ferric, cupric, and other reducible compounds. In the assay
processes, either a standard solution of iodine is employed to
effect a definite chemical reaction or the iodine liberated from an
iodide is determined volumetrically by titration with a suitable
standard solution.
Iodine may function as an oxidizing agent either directly or
indirectly. Thus, when iodine dissolved in potassium iodide
solution reacts with sodium thiosulfate, it acts directly oxidizing
the thiosulfate ion to tetrathionate ion:
or
'Fhe iodine combined as potassium iodide takes no part in the
oxidation, since it is already in ~he ionic state, but the free iodine
is reduced from the molecular-to the ionic state by the gain of
one electron.
Iodine may act indirectly through the formation of hypoiodous
acid which is formed when iodine.is dissolved in water, viz.:
12
+ HOH
HI
155
+ HOI
156
or
and
HaAsOa
+ HOI----7H aAs0 + HI
4
In the last reaction the arsenous acid is oxidized to arsenic

acid by the oxygen made available from water by the reduction
of iodine to the ionic condition. Each of the two atoms of iodine
acquire one electron from one atom of arsenic so that the arsenic
undergoes a change of charge of two when it is oxidized from the
trivalent to the pentavalent state.
The reaction 21-----71 2 can be made to go to completion by
most oxidizing agents under suitable conditions; e.g., when free
chlorine reacts with potassium iodide, each atom of iodine loses
one negative charge or electron and is oxidized from the ionic
to the molecular state:
or
Obviously, the iodine functions as a reducing agent, since each
atom of iodine loses one electron which is gained by an :\.tom of
chlorine.
Since the molecular iodine formed can be determined quantitatively by titration with a standard solution of a reducing agent,
the amount of chlorine which entered into the reaction can be
calculated. See assay of chlorinated lime, page 170.
Potassium iodide when dissolved in an acid solution forms
hydriodic acid and the potassium salt of the acid used; e.g.:
KI
+ HCI----7KCI + HI
When a readily reducible substance is present, it reacts with the

hydriodic acid oxidizing the ionic iodine to the molecular state:
2FeCI s
+ 2HI~2FeCI2 + 12 + 2HCl
or
2Fe+++ + 2I-~2Fe++ + 12
2 X 3( +) + 2( -) = 2 X 2( +)
OXIDATION AND REDUCTION
157
The ferric chloride is reduced at the expense of the iodine.

Since each mole of Fe+++ oxidizes 1 Gm.-atom of iodine to the
molecular state, the amount of Fe+++ which is reduced in a given
reaction can be determined by titrating the iodine oxidized to the
molecular state with a standard solution of a reducing agent
such as sodium thiosulfate. Since the reducing agent removes
the iodine as fast as it is formed, the reaction proceeds to
completion.
Iodometric methods include some of the most exact processes
of volumetric analysis, because, under proper conditions, the
presence of one part of iodine in several million parts of solution
can be recognized by means of starch indicator solution.
Starch Indicator Solution.-This solution is prepared as
follows:
Triturate 1 Gm. of arrowroot starch with 10 cc. of cold distilled
water in a mortar to form a thin paste. Heat about 200 cc.
of distilled water to boiling, add the starch paste to it with constant stirring. Boil the mixture gently for about 30 min. or
until it forms a thin translucent liquid. Since solution of starch
deteriorates rapidly, it should be prepared freshly each day.
Upon hydrolysis in hot aqueous solution starch forms l1-amylose, the soluble portion, and a-amylose, the insoluble portion.
The blue color of starch solution with iodine is thought to be
due to the formation of a colloidal sol made up of l1-amylose,
iodide ion, iodine, and water. Both iodide ion and iodine must
be present fo~ the production of blue color. The blue color is
discharged in the presence of hydroxyl ion if the latter is present
in high concentration. Consequently, titrations with starch
as indicator should not be made in the presence of alkali hydroxides or carbonates, but cold, dilute solutions of alkali bicarbonate
do not affect the end point appreciably.
Standard Solutions.-The standard solutions employed in the
official assays by iodometric methods are 0.1 N iodine and 0.1 N
sodium thiosulfate.
Exercise 48
Object.-To Prepare and Standardize 0.1 N Sodium Thiosulfate.
158
Materials Required.-26 Gm. of sodium thiosulfate.

0.2 Gm. of sodium carbonate.
2 Gm. of potassium iodide.
0.1 N potassium dichromate.
Starch indicator solution.
Procedure.-l. "Dissolve about 26 Gm. of sodium thiosulfate and 0.2 Gm.
of sodium carbonate in 1000 cc. of recently boiled and cooled distilled water."
The thiosulfate is dissolved in water previously boiled to expel

CO 2 , since the latter causes slow decomposition of N a 2S20a which
increases the reducing value of the solution:
Na 2S20a
+ 2H 2CO a
H 2S 20 a
2NaHCO a + H 2S 20 a
H 2SO a + S
As the sulfurous acid forms, the reducing power of the solution becomes. greater, since 1 mole of the sulfurous acid formed
is equivalent to 2 moles of thiosulfate:
2Na2S20a + 12
2Na1 + Na 2S40S
H 2SOa + 12 + H20~2H1 + H 2S0 4
The sodium carbonate acts as a stabilizing agent by converting
the thiosulfuric acid formed into sodium thiosulfate.
2. "Measure accurately 30 cc. of tenth-normal potassium dichromate
into a glass-stoppered flask and dilute it with 50 cc. of distilled water. Add
2 Gm. of potassium iodide and 5 cc. of hydrochloric acid, stopper and allow
to stand for ten minutes. Dilute with 100 cc. of distilled water and titrate
the liberated iodine with the sodium thiosulfate solution. When the solution has assumed a yellowish-green color, add starch T.S. and continue with
the titration to the discharge of the blue color. Calculate the normality of
the sodium thiosulfate solution and, if desired, adjust exactly to tenthnormal."
The acid solution of potassium iodide quantitatively reduces

chromic acid to green chromic chloride, liberating an equivalent
quantity of iodine:
K 2Cr 207 + 6KI +
294.2 6(166.02)
14HCl~2CrCla
+ 8KCl + 7H 20 + 31 2.
6(126.92)
It is evident from the above equation that 1 mole, 294.2 Gm.,

of K 2Cr 207 reacts with 6 moles, 996.12 Gm., of K1 to oxidize
761.52 Gm.-atoms of iodine from the ionic to the molecular state.
159
Potassium iodide must be present in excess to prevent the

precipitation of iodine which is insoluble in aqueous solution but
soluble in KI solution.'
The solution is diluted to a large volume, 350 cc., to make the
end point more easily distinguishable, for the latter is not indicate~ by the disappearance of color but by a change in color
from blue to light green.
Since tenth-normal oxidizing and reducing solutions are equivalent cubic centimeter for cubic centimeter, the calculation of
the normality of the thiosulfate solution is as follows:
cc. 0.1 N K 2 Cr 207 X 0 1 = normality of the sodium thiosulfate
.
cc. N a 2S20a
solution.
Sodium thiosulfate solution changes in its reducing power
due to the action of carbon dioxide, sunlight, and dust. The
solution should, therefore, be preserved in tightly stoppered
bottles and be kept away from direct sunlight.
Sodium thiosulfate solution may also be standardized against
pure iodine dissolved in potassium iodide solution or against
iodine set free from acidulated solution of potassium iodide by
standard solutions of potassium permanganate, potassium bromate, or potassium iodate.
1. Illustrate how iodine may ~unction directly and indirectly as an oxidizing agent.
2. How may CO 2 change the reducing power of standard sodium thiosulfate solution? Use reactions to illustrate the explanation.
3. When potassium dichromate is reduced by iodine, what change in
polarity does each Cr atom undergo? Is the iodine oxidized or reduced?
4. Why must KI be present in excess before the K 2Cr 207 is added?
6. What purpose does the HCI serve?
Exercise 49
Object.-To Prepare 0.1 N Ionine Solution.

Materials Required.-13 Gm. of pure iodine.
36 Gm. of potassium iodide .
Procedure.-"Weigh accurately about 12.75 Gm. of reagent iodine and
transfer it quickly into a solution of 36 Gm. of potassium iodide in 100 cc.
of distilled water. After solution is complete, dilute to exactly 1000 cc. at
160
25C. From the weight of the iodine used calculate the normality.
normality of the solution should frequently be redetermined."
The
One gram of iodine is soluble in 2,950 cc. of water. In the

presence of sufficient potassium iodide, however, its solubility
is greatly increased due to the formation of an unstable, soluble
polyiodide of potassium, KIa:
K1
+ 12
KIa
In the presence of reducing agents, the reaction proceeds quantitatively to the left, and the solution reacts like a solution of
iodine alone.
The iodine should be weighed accurately in a glass-stoppered
weighing bottle, transferred quantitatively to the solution of
36 Gm. of KI in 100 cc. of distilled water, and after it is completely dissolved, the soll!tion should be made up to exactly
1,000 cc. The solution of potassium iodide used to dissolve
the iodine is of sufficient concentration to dissolve the iodine
rapidly so that no appreciable loss will occur due to volatilization.
If pure reagent iodine is used in preparing this solution, it will be
approximately tenth-normal and need not be standardized. Due,
however, to the difficulty of weighing exact quantities of iodine,
it is usually more convenient to prepare a solution of app~oximate
strength and standardize it against 0.1 N sodium thiosulfate
solution as directed in the following exercise.
Exercise 50
Object.-To Prepare and Standardize 0.1 N Iodine against

Sodium Thiosulphate.
Materials Required.-14 Gm. of pure iodine.
0.1 N sodium thiosulfate,
Starch test solution.
Procedure.-Dissolve about 14 Gm. of reagent iodine in a solution of
36 Gm. of potassium iodide in about 100 'cc. of distilled water, dilutingfinally to 1,000 cc. Carefully measure from a burette 25 cC. of this solution
into a flask, then add gradually and cautiously from a burette 0.1 N sodium
thiosulfate, shaking constantly, until the color of the solution is but slightly
yellow, then add a few drops of starch T.S. and continue the addition of the
0.1 N sodium thiosulfate until the blue color is just discharged. Note the
number of cubic centimeters of the 0.1 N sodium thiosulfate consumed, and
161
then dilute the iodine solution so that 25 cc. will require for decolorization
exactly 25 cc. of the 0.1 N sodium thiosulfate at standard temperature.
Although the U.S.P. directs that reagent iodine be used in

preparing 0.1 N iodine solution, iodine containing chlorine in
small amounts as impurity could be used, since the chlorine
reacts'with potassium iodide to liberate iodine:
,ICI
+ KI
KCl
+ 12
The sodium thiosulfate reacts with the iodine to form sodium

iodide and sodium tetrathionate, and a very slight excess of
thiosulfate discharges the blue color of the indicator:
12
+ 2Na2S20a
2NaI
+ Na2S406
If 25 cc. of approximately 0.1 N iodine require exactly 25 cc.

of 0.1 N sodium thiosulfate, the iodine is exactly tenth-normal,
but,' if only 24 cc. of 0.1 N iodine solution are required, the
solution is stronger than normal and each 24 cc. of iodine solution must be diluted to 25 cc. to secure an exactly 0.1 N solution.
Since 0.1 N iodine solution deteriorates, it will not remain
precisely tenth-normal very long. It is therefore best to calculate
the normality of the solution and employ it as a standard solutIon
of known value. In the above case, the approximately 0.1 N
iodine solution would be 25/24 = 1.0417 times tenth-normal
and should be labeled 0.1042 N iodine. Whenever the solution
is used in a determination, the number of cubic centimeters of
0.1 N iodine employed may be calculated; i.e., cubic centimeters
approximately 0.1 N iodine X 1.0417 = cubic centimeters
0.1 N iodine.
Iodine solutions may also be standardized against pure arsenous
oxide (see Exercise 51).
1. Why may iodine containing chloriJ;le or bromine as impurity be used to
prepare standard solutions of iodine when the solution is standardized
against sodium thiosulfate?
2. What is the normality of a solution containing 1.5420 Gm. of iodine
in 100 cc.?
3. If 24.25 cc. of iodine solution requires 25 cc. of 0.0858 N Na 2S20S. what
is the normality of the iodine solution?
162
OXIDATION-REDUCTION METHODS BY DIRECT TITRATION WITH

STANDARD IODINE SOLUTION
Exercise 51
Object.-Assay of Arsenic Trioxide.

Materials Required.-About 0.2 Gm. of arsenic trioxide.
Sodium hydroxide test solution prepared by dissolving 4.3 Gm. of NaOH
in sufficient distilled water to make 100 cc.
Diluted sulfuric acid.
2 Gm. of sodium bicarbonate.
0.1 N iodine.
Procedure.-l. Dissolve about 0.2 Gm. of arsenic trioxide previously
dried to constant weight at 100C., and accurately weighed, in 20 cc. of
boiling distilled water, and gradually add sodium hydroxide T.S. until
complete solution results.
Arsenic trioxide is slowly soluble in cold water, more rapidly

soluble in boiling water, and readily soluble in sodium hydroxide
solution with the formation of sodium arsenite:
As 20 a T 6NaOH
197.82
2Na aAsOa
+ 3)I 20
Sodium arsenite on hydrolysis yields sodium hydJoxide and

arsenous compounds to form a distinctly alkaline 'solution.
2. Neutralize this solution with diluted sulfuric acid, using phenolphthalein T.S. as indicator, cool, dissolve in it 2 Gm. of sodium bicarbonate,
and titrate the mixture with 0.1 N iodine, using starch T.S. as indicator.
Each cubic centimeter of 0.1 N iodine corresponds to 0.004946 Gm. of
As 20 .
If iodine is added to this alkaline solution, some of it tends to

form sodium hypoiodite, NaIO, or similar compounds which do
not react readily with arsenous acid:
2NaOH
+I
-NaIO
+ NaI + H
The excess sodium hydroxide, therefore, required to dissolve the

As 20 a is exactly neutralized by sulfuric acid. Sodium bicarbonate is added to neutralize the HI formed in the reversible reaction
163
The NaHCO a removes the HI as rapidly as it is formed, causing

the reaction to go to completion toward the right:
Under the conditions of the assay, sodium bicarbonate does

not react with iodine, but, if an excess of iodine is added, it may
react slightly. The reaction between arsenous (wid and iodine
proceeds quantitatively to completion best in a neutral solution.
Since HI is formed in the reaction, a buffer must be added to keep
the solution neutral. The sodium bicarbonate in this case is
the buffer.
The oxidation of a mole of arsenic from As+++ to As+++++
requires 2 Gm.-atoms of iodine. The change of polarity of
As+++ when oxidized to As+++++ is 2( +), and since each mole of
As 20 a contains 2 atoms of As, the total change of polarity is
2 X 2( +) = 4(+). One cubic centimeter of 0.1 N iodine is
therefore equivalent to 4 X
~~7:;,000 =
0.004946 Gm. As 2 0 a, or
74.96
2 X 10 X 1,000 = 0.003738 Gm. As.
U.S.P. arsenic trioxide should contain 99.8 per cent of As 20 a

Calculate the per cent purity of the As 20 a assayed and the per
cent of arsenic which it contains. Many arsenous compounds,
antimonous compounds, etc., and their preparations may be
assayed by similar procedure.
1. Write the reaction between As 20 a and NaOH structurally.
2. Why must the excess sodium hydroxide be neutralized with acid before
the titration with standard iodine solution?
3. What substance is employed as a buffer in the assay? Why is a buffer
necessary?
4. Name some other official substa'nces or preparations containing antimonous or arsenous compounds to which the above method is applied.
5. If pure As 20 a is available, how might a solution of iodine be standardized against it?
6. Ascertain the official method of assay for antimonous oxide, explain
each step in the procedure, write equations for the reactions involved, and
calculate the equivalent weight of 8b 20 a
164
TABLE XVII1.-0FFICIAL SUBSTANCES ASSAYED BY DIRECT TITRATION WITH

0.1 N IODINE
Amount
Equivaused,
lent of
Gm. or
1 cc.
cc.
Substance
per cent
--U.S.P.
Antimony and potassium tartrate ....
0.5
Arsenic triiodide .................
0.5
Arsenic trioxide ...
0.2
.......
Arsenous acid, solution .............. 20
Potassium arsenite, solution.
.... 20
Sodium thiosulfate ..... .... ....
0.5
Tryparsamide .........
. . . . . . . . . . 0.2
N.F.
Arsenic and mercuric iod.ide, solution
(for AsI,) ........................ 25
Sodium cacodylate, ampuls ... .......
0.2"
Sodium thioBulfate, ampule ...........
I"
0.01670
0.02278
0.004946
0.004946
0.004946
0.01581
0.003746
K(SbO)C.H.O,.UH,O = 99
AsI, = 99
As,O, = 99.8
As,O, = 0.975 to 1.025W/V
As,O, = 0.975 to 1.025W/V
Na,S,O, = 99
As = 25.1 to 25.5
0.02278 AsIa = 0.95 to 1.05W/V

0.007998 Na(CH,),AsO, = 69 to 76'
0.01581 Na,S,O, = 60.5 to 66.9'
" Amount of ingredient sought.

OXIDATION-REDUCTION METHODS BY DIRECT TITRATION OF

FREE IODINE WITH STANDARD SODIUM THIOSULFATE
SOLUTION
Exercise 52
Object.-Assay of Compound Solution of Iodine for Iodine

Content.
Materials Required.-5 cc. of compound solution of iodine.
0.1. N sodium thiosulfate.
Procedure.-<iDilute exactly 5 ce. of Compound Solution of Iodine with
25 cc. of distilled water, and titrate with tenth-normal sodium thiosulfate,
starch T.S. being used as the indicator. Each cubic centimeter of tenthnormal sodium thiosulfate is equivalent to 0.01269 Gm. of 1."
The compound solution of iodine is diluted with water to bring

it to approximately tenth-normal. This dilution can be made
without the addition of K1 to prevent precipitation of iodine,
since the compound solution of iodine contains 10 per cent of KI.
The starch solution should not be added until the titration mixture has acquired a straw color. The following reaction takes
place:
12
2Na2S203.5H20~Na2S406
2(126.92)
+ 2NaI + 10H20
165
Each cubic centimeter of 0.1 N sodium thiosulfate consumed

in the titration is equivalent to 2 X
~~3~\000 = 0.012692 Gm.
iodine. The U.S.P. requires that compound solution of iodine

contain not less than 4.5 or more than 5.5 Gm. of iodine per
100 cc. of solution. Note that this is a weight-to-volume percentage. Calculate the per cent of iodine in the sample assayed
and compare the result with the official requirement.
Elemental iodine, tinctures of iodine, and any preparation
containing free iodine may be assayed by methods similar to
that given above, provided that other substances which oxidize
sodium thiosulfate are absent.
1. Why can compound solution of iodine be diluted without adding KI
to prevent the precipitation of iodine?
~. Give a procedure for the assay of elemental iodine with reasons for
each step. Write equations for all of the reactions involved.
3. Name several substances which, if present, would oxidize sodium
thiosulfate.
4. Why is it best not to add the starch indicator solution before the iodine
solution has acquired a straw color?
TABLE XIX.-OFFICIAL SUBSTANCES ASSAYED BY DIRECT TITRATION WITH
0.1 N Na 2S 20a
Slfbstance
U.S.P.
Iodine ......................
Iodine, compound solution of.
Iodine pentoxide, R ..........
Iodine, tincture of ...........
Iodine, tincture of, mild ......
N.F.
~odine, ampuls of. ...........
Iodine, stronger tincture of ...
Iodic acid, R ................
R.
= reagent.

b Per cent of the labeled amount.
a
Amount
Equivaused,
lent of
Gm.
1 cc.
or cc.
per cent
12 =
12 =
1 20.
12 =
12 =
0.5
5.0
0.5
5.0
5.g
0.01269
0.01269
0.002782
0.01269
0.01269
o. 175a
0.01269 12 = 95 to lO5W IVb

0.01269 12 = 16 to 17WIV
0.002932 HIO s = 99.8
2.0
0.1
99.5
4.5'to 5.5W IV
= 99.5
6.5 to 7.5W IV
1.8 to 2.2W IV
166
OXIDATION-REDUCTION METHODS INVOLVING THE USE OF

EXCESS STANDARD IODINE SOLUTION AND RESIDUAL TITRATION OF THE EXCESS IODINE WITH STANDARD SODIUM
THIOSULFATE SOLUTION
Exercise 53
Object.-Assay of Mercurous Chloride.

Materials Required.-About 0.7 Gm. of mercurous chloride.
50 cc. of 0.1 N iodine.
0.1 N sodium thiosulfate.
Procedure.-l. "Dry about 0.7 Gm. of Mild Mercurous Chloride to
constant weight over sulfuric acid, weigh accurately, mix well with 10 cc.
of distilled water in a glass-stoppered flask, and add 50 cc. of tenth-normal
iodine and 5 Gm. of potassium iodide dissolved in 10 cc. of distilled water."
Since mercurous chloride is practically insoluble in water, the

purity of the salt cannot be determined by the precipitation
method used to determine the chloride ion in a soluble chloride.
When mercurous chloride is treated with a solution of iodine in
potassium iodide, it dissolves quite readily with the formation
of a double salt of mercury and potassium iodides:
2HgCl
236.07
+ 6KI + I -,)2HgK I + 2KCl

2
2 4
2. "Stopper the flask, allow the mixture to stand with occasional agitation
until complete solution has taken place, and then titrate the residual iodine
with tenth-normal sodium thiosulfate, starch T.S. being used as indicator.
Each cubic centimeter of tenth-normal iodine is equivalent to 0.02361 Gm.
of HgCl."
In the assay process, the mercurous chloride mixture is allowed

to stand with occasional agitation until the reaction has gone to
completion as indicated by its complete solution. The excess
0.1 N iodine added is then determined by residual titration. The
formula of the double salt of mercuric potassium iodide may be
written, HgI 2 .2KI, and the ionic reaction may be represented as:
2Hg+
+ k-~2Hg+t- + 21-
The number of cubic centimeters of 0.1 N iodine found upon

residual titration with 0.1 N sodium thiosulfate subtracted from
the number of cubic centimeters originally added gives the
167
volume of standard iodine solution consumed in the oxidation of

the mercurous ion to mercuric ion. Since 1 Gm.-ion of iodine is
equivalent to 1 Gm.-mole of HgCI, each cubic centimeter of
0.1 N iodine consumed is equivalent to 10 2~61~~00 = 0.02361
Gm. HgCl.
1. Why is the mixture of HgCI with solution of iodine in KI stoppered
and allowed to stand?
2. Indicate by ionic reactions how iodine functions as an oxidizing agent
in the assay of sodium sulfite (U.S.P. reagent) and sulfurous acid (U.S.P.
reagent).
3. Explain each step involved in the official assay of mercurous iodide
and write ionic ally all of the reactions which occur during the assay.
4. If an unknown sample treated with 50 cc. of 0.950 N iodine required
18.6 of 0.1025 N sodium thiosulfate to titrate the excess iodine, to what
amount of each of the following substances would the sample be equivalent:
CH~COCH" HgCI, Na 2S.9H20, and S02?
TABLE XX.-OFFICIAL .sUBSTANCES ASSAYED BY RESIDUAL TITRATION OF

ExCESS STANDARD IODINE SOLUTION WITH 0.1 N Na 2S 20,
Substance
Amount
used,
Gm. or
Equivalent of
1 cc.
CC.
U.S.P.
Acetone ..................
Mercurous chloride ........
Mercurous iodide ..........
Methylthionine chloride ...
Sodium bisulfite, R ........
Sodium sulfite, R ..........
Sulfurous acid, R ..........
l.0
0.7
1.0
0.12
....
0.25
2.0
per cent
---
0.0009675
0.02361
0.03275
0.005328
0.005203
0.006303
0.003203
CR,COCR, = 99
HgCI = 99.6
HgI = 99
C 16H 1S N 3C1S = 98.5
NaHSO, = 90
Na 2S03 = 95
S02 = 6
N.F.
Mercurous chloride, mild,
and sodium bicarbonate,
tablets of ...............
Mercurous chloride, mild,
tablets of ...............
R.
= reagent .
Amount of ingredient sought.

, Per cent of the labeled amount.
0.3" '" 0.02361
HgCI = 92.5 to 107.5b
0.02361
HgCl = 92.5 to 107.5b
0.3"
168
OXIDATION -REDUCTION METHODS INVOLVING THE LIBERATION

OF IODINE FROM POTASSIUM IODIDE AND TITRATION OF
THE FREE IODINE WITH STANDARD SODIUM
THIOSULFATE SOLUTION
Exercise 54
Object.______:_Assay of Solution of Ferric Chloride.

Materials Required.-About 2 Gm. of solution of ferric chloride.
Procedure.-l. Transfer about 2 Gm. of a solution of ferric chloride to
a tared flask, stopper, and weigh accurately; add 5 cc. of hydrochloric acid,
25 cc. of distilled water, and about 3 Gm. of potassium iodide. Securely
stopper the flask, and allow the mixture to stand for 5 min.
The solution of ferric chloride may more conveniently be

weighed in a glass-stoppered weighing bottle. Loosen the stopper
of the weighing bottle and place the bottle, stopper, and contents
in a 250 cc. Erlenmeyer flask containing 5 cc. of HCI and 25 cc.
of distilled water. Remove the stopper from the weighing bottle
with a stirring rod and thoroughly mix the solutions, add the
KI, and stopper the flask securely. The solution is l allowed to
stand for 5 min. to permit the following reactions to go to
completion:
KI
HCl----+ KCI
HI
2FeCI s
2HI----+2FeCb
162.21
+ 2HCl + 12
or
2Fe+++
+ 2I---+2Fe++ + 12
Sufficient KI must be present to form enough HI to reduce the

ferric ion quantitatively to ferrous ion and to dissolve the free
iodine formed.
2. Dilute the liquid, with 50 cc. of distilled water, and then titrate the
liberated iodine with 0.1 N sodium thiosulfate, starch T.S. being used as
indicator. Each cubic centimeter of 0.1 N sodium thiosulfate corresponds
to 0.005584 Gm. of Fe.
169
The liberated iodine is titrated with 0.1 N sodium thiosulfate

in the usual manner.
12
+ 2Na2S20g.5H20~2Na1 + Na2S406 + 10H20
Each mole of Fe+++ is equivalent to 1 Gm.-atom of iodine.

Each cubic centimeter of 0.1 N Na 2S20g is therefore equivalent
162.21
55.84
to 10 X 1,000 = 0 016221 Gm. FeCla or 10 X 1,000 = 0.005584
Gm.Fe.
According to the U.S.P. definition, solution of ferric chloride
should contain not less than 10 per cent or more than 11 per cent
of Fe. Calculate the per cent of Fe and the per cent of FeCla
in the sample assayed and compare the results obtained with the
Fe BY TITRATION
0.1 N Na 2S 20a
TABLE XXI.-OFFICIAL SUBSTANCES ASSAYED FOR

IODINE LIBERATED FROM KI WITH
Amount Equivaused,
lent of
Gm. or
1 cc.
cc.
Substance
---------
~---I---
U.S.P.
}'erric
Ferric
Ferric
Ferric
chloride, solution of .. .......... .

chloride, tincture of. ..........-' .
citrate, R . ................... .
sulfate. solution of ............. .
Iron and ammonium nitrates ..... ... , .
Iron and ammpnium citrates, green . .. .
2.0
5.0
1
1.5
1.0
1
OF THE
Official requirement.
per cent
---1-------0.005584
0.005584
0.005584
0.005584
0.005584
0.005584
Fe
Fe
Fe
Fe
Fe
Fe
= 10 to 11
= 4.5
= 16.5 to 18.5
= 9.5 to 10.5
= 16.5 to 18.5
= 14.5 to 16
0.005584
0.005584
0.005584
0.005584
0.005584
0.005584
0.005584
0.005584
Fe = 4.48 W IV
Fe = 17
Fe = 21.8
Fe = 2.8 to 3.2
Fe = 12 to 15
Fe = 10.5 to 12.5
Fe = 20 to 22WIV
Fe = 0.16 to 0.20W/V
N.F.
Ferric citrochloride. tincture of. .... "
5.0
Ferric glycerophosphate .............. .
0.5
Ferric hypophosphite ................. .
1.0
Ferric oxide, saccharated .............. .
8.0
Ferric phosphate, soluble . ............ .
1.0
Ferric pyrophosphate, soluble. . . . .
..
1.0
Ferric subsulfate, solution of ......... .
1.0
Iron. and ammonium acetate, solution of.
25
Iron and ammonium citrates, green,
ampuls of .......................... '" I"
Iron, peptonized. . .. . .......... , .... .
0.5
t
0.005584 Fe
0.005584 Fe
= 14.5 to 16.0b
= 16 to 18
Iron, peptonized, and manganese, solu-
tion of .......................... .
Iron, peptonized. solution of . .......... .
R. = reagent.
25
25
0.005584 Fe = 0.265 to 0.325W/V

0.005584 Fe = 0.265 to 0.325WIV
170
The principle involved in the method of assay in this exercise is

applicable to most official determinations of ferric salts with slight
modifications, e.g., solution of iron and ammonium acetate,
solution of ferric sulfate, soluble ferric phosphate, solution of
ferric citrate, albuminized iron, peptonized iron, ferrous lactate,
and others.
1. Explain how potassium iodide in acid solution functions as a reducing
agent.
2. Ascertain the method of assay of peptonized iron in the N.F. and
apply the principles studied in the above assay in an explanation of each
step of the procedure.
3. Calculate the amount of ferric sulfate, ferric oxide, and ferrous lactate
equivalent to 1 cc. of 0.1 N KMn04.
Exercise 55
Object.-Assay of Chlorinated Lime.

Materials Required.-About 4 Gm. of chlorinated lime.
5 cc. of acetic acid.
Procedure.-l. "Transfer to a mortar about 4 Gm. of ChldrinatedLime,
accurately weighed in a tared weighing bottle, using 50 cc; of distilled water.
Triturate thoroughly, and pour the mixture into a 1,000 cc. graduated flask,
rinsing the mortar with distilled water to make 1,000 cc. Stopper the flask
and allow it to stand for ten minutes."
The accurately weighed sample of chlorinated lime is triturated

with water to disintegrate it to a fine powder.
2. "Shake thoroughly, add to 100 cc. of the mixture 1 Gm. of potassium
iodide and 5 cc. of acetic acid, and titrate the liberated iodine with tenthnormal sodium thiosulfate, starch T.S. being used as the indicator. Each
cubic centimeter of tenth-normal sodium thiosulfate is equivalent to 0.003546
Gm. of available chlorine (CI)."
An aliquot portion of the mixture representing 0.4 Gm. of

sample is treated with 1 Gm. of KI and 5 cc. of acetic acid. In
the presence of acids the available chlorine is liberated from the
chlorinated lime. Available chlorine is usually defined as the
chlorine that is liberated from a substance by acids. The free
171
chlorine reacts with K1 to liberate iodine quantitatively, and

the quantity of iodine set free is determined by titration with
0.1 N N a2S20a.
Chlorinated lime is thought to be a calcium chloro-hypochlorite,
Ca(OCI)CI, which may be regarded as a compound containing
equ~molecular portions of calcium hypochlorite, Ca(OCI) 2, and
calcium chloride, GaCI 2:
2Ca(OCI)CI--CaCb
+ Ca(OClh
All of the chlorine present in calcium chloro-hypochlorite

released when the compound is treated with acetic acid:
Ca(OCI)CI
IS
+ 2CH aCOOH ~Ca(CHaCOOh + H 20 + Cb
The liberated Cl 2 reacts with K1:

Cl z + 2K1--2KCI + 12
2(35.46)
2(126.92)
The iodine set free from K1 is titrated with 0.1 N Na 2S20a:
12
+ 2Na 2S20a.5H 20-2Na1 + Na 2S40S + 10H 20
From the above reactions, it is apparent that 2N a 2S20a is equivalent to 12 which is equivalent to Ch
One cubic centimeter of 0.1 N Na 2S20a is equivalent to
10 1~6i~~00 = 0.012692 Gm. iodine and 10 ~i~oo = 0.003546
Gm. chlorine.
Other substances which yield free chlorine or free bromine
when treated with acids may be assayed in the same way, e.g.,
solution of sodium hypochlorite.
1. What is "available chlorine"?
2. What function does KI serve in"this assay? Must the KI be present
in excess?
3. If a 0.4 .Gm. sample is known to contain 32 per cent of available
chlorine, how much KI would be necessary to yield an amount of iodine
equivalent to the chlorine?
4. Look up the U.S.P. requirement for chlorinated lime and compare it
with the assay result.
172
TABLE XXII.-OFFICIAL SUBSTANCES ASSAYED FOR AVAILABLE CHLORINE

BY TITRATION OF THE IODINE LIBERATED FROM KI WITH
Amount
Equivaused,
lent of
Gm. or
1 cc.
cc.
Substance
U.S.P.
Chloramine-T ..........
Dichloramine-T ........
Chlorinated lime, R .....
Sodium hypochlorite,
solution of ...........
Sodium hypochlorite,
solution of, diluted ...
0.1 N Na 2S20.
per cent
0.2
0.1
4
0.001773 Cl (active) = 11.5 to 13

0.001773 Cl (active) = 28 to 30
0.003546 Cl (available) = 30
0.003723 NaOCI = 4 to 6
25
0.003723 NaOCl = 0.45 to 0.50W IV
R. = reagent.
Exercise 56
Object.-Assay of Cupric Sulfate.

Materials Required.-1 Gm. of cupric sulfate.
4 cc. of acetic acid.
Procedure.-"Dissolve about 1 Gm. of Cupric Sulfate, \ accurately
weighed, in 50 cc. of distilled water, add 4 cc. of acetic acid and 3 Gm. of
potassium iodide, and titrate the liberated iodine with tenth-normal sodium
thiosulfate, starch T.S. being used as indicator. Each cubic I centimeter
of tenth-normal sodium thiosulfate is equivalent to 0.01596 Gm. of CUS04."
This assay is based on the reaction between cupric sulfate

and potassium iodide in which the copper is precipitated as
cream-colored cuprous iodide and one atom of iodine is liberated
for each cupric ion present:
2CUS04 + 4KI~Cu212
2(159.63)
or
2Cu++
+ 2K2S0 + 12
4
+ 4I-~2Cu+ + 21- + 12
Each cubic centimeter of sodium thiosulfate consumed is

126.92
159.63
equivalent to 10 X 1,000 = 0.01269 Gm,. 12 , to 10 X 1,000
63.57
= 0.01596 Gm. CUS04, and to 10 X 1,000 = 0.00636 Gm. Cu.
173
U.S.P. copper sulfate should contain not less than 63 per cent
or more than 66.8 per cent .of CUS04 corresponding to 98.5
per cent of CuS04.5H20. Calculate the per cent Cu, CUS04
and CuS04.5H 20 in the sample assayed.
1. What function does the acetic acid serve in this assay?
2. Explain the reduction of copper from the cupric to the cuprous condition in accordance with the ionic theory. Indicate all transfers of charge.
3. How much iodine will 0.25 Gm. of CUS04 liberate from KI in acid
solution?
Exercise 57
Object.-Assay of Exsiccated Sodium Arsenate.

Materials Required.--o.3 Gm. of exsiccated sodium arsenate .
.10 cc. of hydrochloric acid.
Procedure.-"Dissolve about 0.3 Gm. of Exsiccated Sodium Arsenate,
dried to constant weight at 150C. and accurately weighed in 25 cc. of distilled water in a glass-stoppered flask; heat the solution to 80C., and add
10 cc. of hydrochloric acid and 3 Gm. of potassium iodide. Allow the mixture to stand in the stoppered fla:sk for 15 minutes at 80C.; then cool, and
titrate the liberated iodine with tenth-normal sodium thiosulfate, using
starch T.S. as the indicator. Conduct a blank test, and deduct the amount
of tenth-normal sodium thiosulfate consumed in the blank. Each cubic
centimeter of tenth-normal sodium thiosulfate is equivalent to 0.009295 Gm.
of Na 2 HAs0 4."
Sodium arsenate in strongly acid solution liberates iodine from

KI with reduction of As+++++ to As+++, the reaction going to
completion when the solution is warmed and allowed to stand.
The reactions which take place, although probably more complex,
may be represented as follows:
"
Na 2HAs0 4 + 2HCI~H3As04 + 2NaCI

185.91
HCI + KI~KCl + HI
Ha As04 + 2H1~H3As03 + H 20 + 12
2Na2S20a + 12~2Na1 + Na 2S406
174
Since each mole of Na 2HAs0 4 liberates 2 Gm.-atoms of iodine,
. 1
185.91
1 cc. o
f 0
. 1 N N a2S 2O3 IS eqmva ent to 2 X 10 X 1,000
0.009295 Gm. Na2HAs04.
N.F. exsiccated sodium arsenate contains not less than 98
per cent of Na 2HAs0 4 Calculate the per cent purity of the
sample assayed and compare the result with the official requirement. Also calculate the per cent of arsenic in the sample
assayed.
Arsenic and antimonic compounds may be assayed by methods
similar to that given above. The arsenic and antimony present
in organic compounds in the trivalent state may also be assayed
by this method after quantitative oxidation to the pentavalent
state. Such procedure is employed in the official assays of
arsphenamine and neoarsphenamine.
1. Assuming that the sample used in the assay was pure Na.HAsO . 7H.O,
how much KI would be required?
_..2. Write the equations involving oxidation-reduction ionically.
3. Name several official arsenic and antimonic compoundi assayed by
methods which employ the same basic principles.
4. What change in valence does the arsenic undergo in the above assay?
Does the change of valence correspond with the change of charge?
TABLE
X 4 III.-OFFICIAL
SUBSTANCES CONTAINING ARSENIC ASSAYED BY
TITRATION OF THE IODINE LIBERATED FROM
Substance
Amount
Equivaused,
lent of
Gm. or
1 cc.
cc.
KI
WITH
0.1 N Na 2S20.
per cent
-U.S.P.
Arsphenamine .....
Neoarsphenamine ..
N.F.
Sodium arsenate,
exsiccated .......
Sodium arse'nate,
solution of. ......
0.2
0.2
0.003746 'As = 30
0.003746 As = 19 to 22
0.3
0.009295 Na 2HAsO. = 98
25
0.009295 Na 2HAsO. = 0.975 to 1.025W IV
175
Exercise 68
Object.-Assay of Thyroid.
Materials Required.-1 Gm. of thyroid.
A nickel crucible.
9 Gm. of anhydro-qs potassium carbona,te.
7 Gm. of anhydrous sodium carbonate.
5 Gm. of potassium nitrate.
50 cc. of solution of chlorinated soda.
About 60 cc. of diluted phosphoric acid (1 to 1).
0.1 Gm. of potassium iodide.
Procedure.-l. "Thoroughly mix 1 Gm. of Thyroid, finely powdered and
accurately weighed, with 15 Gm. of an intimate mixture of 138 parts by
weight of anhydrous potassium carbonate, 106 parts of anhydrous sodium
carbonate arid 75 parts of powdered potassium nitrate in a nickel crucible
of about 125-cc. capacity, and spread an additional 5 Gm. of this mixture
evenly over the surface. Heat the crucible with the flame of a Bunsen
burner at such a rate as to attain a dull red color in ten minutes, and continue
the heating at the same temperature for an additional ten-minute period.
At the end of this time the carbonaceous material is completely oxidized
and the mixture has just begun to melt around the wall of the crucible.
Cool the fusion mixture and place the crucible and contents in a 400-cc.
beaker. Add 150 cc. of hot distilled water and stir until the contents of the
crucible are completely dissolved."
Dried thyroid gland contains iodine in organic combination.

The organic matter is oxidized by a fusion mixture containing
alkali carbonates, which serve to fix the liberated iodine in the
form of iodide, and potassium nitrate, which acts as an oxidizing
agent. The carbonates used in the fusion mixture must be
anhydrous to prevent loss of material by decrepitation. A layer
of fusion mixture is spread over the surface of the mi~ture in the
crucible as a precaution against the loss of iodine. When this
mixture is heated a number of reactions undoubtedly take place,
but the only ones which pertaiJt to the assay for iodine may be
regarded as:
2RI + K2C03~2KI + CO 2 + H 20
KI + 3KN03~KI03 + 3KN0 2
In the above reactions it is assumed that all of the oxygen set
free when potassium nitrate is heated is not utilized in the oxida-
176
tion of the organic matter present but that some of it reacts with
KI to form KIO g After carbonization is complete and the
crucible has cooled, the residue is treated with distilled water
which dissolves the iodide, iodate, carbonates, and nitrite.
2. "Transfer the solution to a 500-cc. Erlenmeyer flask and rinse the
beaker and crucible with four, lO-cc. portions of hot distilled water, adding
the rinsings to the solution in the flask. Cool the solution to 15C. and
add 50 cc. of freshly prepared chlorinated soda T.S. Cautiously add 60 cc.
of dilute phosphoric acid (made by mixing equal volumes of phosphoric acid
and distilled water), place the flask on a hot plate and boil the solution until
a strip of filter paper, moistened with starch-potassium iodide T.S., does not
become blue when held in the vapor in the mouth of the flask. The final
volume of solution in the flask must be about 175 cc., and distilled water
must be added, if necessary, during the boiling to maintain this volume."
Solution is best effected by treating the residue with successive

portions of warm distilled water. The combined solution is
treated with solution of chlorinated soda which functions as an
oxidizing agent to convert any KI present in the solution into
KIO a. The oxidation with sodium hypochlorite N aOOl is carried
out in solution acidulated with phosphoric acid. The reactions,
although more complex, may be represented 8,s:
2NaOOI + H aPOc--tNa 2HP0 4
KI + 3HOOI----tKIO g + 3HOl
HOI + HOOl----tHOH + Ob
+ 2HOCI
Excess phosphoric acid is added to liberate all of the combined

chlorine present in the N aOOl. The solution is boiled to drive
off all free chlorine, since chlorine if present would liberate i9dine
from KI.
3. "Cool the solution to about 25C. and add 10 cc. of a freshly prepared
aqueous solution of potassium iodide (1 in 100). Titrate the liberated
iodine with two-hundredth-normal sodium thiosulfate, adding 3 cc. of
arrowroot starch T.S. as indicator just before the end of the titration. Conduct a blank test with the liIame quantities of the same reagents, omitting
only the Thyroid, and fusing as directed, and subtract the volume of twohundredth-normal sodium thiosulfate consumed from that consumed by the
Thyroid. Each cubic centimeter of the corrected volume of two-hundredthnormal sodium thiosulfate is equivalent to 0.0001058 Gm. of I."
When the cool soluti.on is treated with KI in acid solution,

iodine is set free quantitatively by the iodate:
177
2Kl
oHl
+ H P0 K HP0 + 2Hl
+ K10a~Kl + 3H20 + 31
3
The liberated iodine is titrated with 0.005 N Na2S20a:
Since each mole of KIOa.obtained from the thyroid liberates

6 Gm.-atoms of iodine from Kl, each cubic centimeter of 0.005
N Na 2S 20a is equivalent to 6 X
i02;.~21,000
0.0001058 Gm.
iodine derived from thyroid glands.

Thyroxin and thymol iodide are assayed by modifications of
the above procedure.
1. What purpose is served by the alkali carbonates in the fusion mixture?
2. How does the solution of chlorinated soda function as an oxidizing
agent?
3. Why must all of the liberated chlorine be expelled previous to the
addition of KI? Indicate by an equation what would occur if the chlorine
was not driven off.
4. When KIO, is reduced by HI, what change of charge does the iodine
combined as iodate undergo? Does the change of charge correspond with
the change of valence?
D. How may the use of 0.005 N Na 2S 20" in place of 0.1 N Na 2S203 commonly used, minimize error in the measurement of volume?
TA"BLB XXIV.--o"FFIClAL SunSTANcBS ASE.AYBD FOR IODINE "BY CONVERSION
OF THE IODINE TO IODATE ANIl TITRATION OF THE IODINE LIBERATED
FROM
Substance
KI
WITH STANDARD
Na 2S 20.
Amount
EquivaNorused,
malityof lent of
Gm. or
1 cc.
Na 2S20.
cc.
U.S.P.
Calcium iodobehenate ..
Thymol iodide .........
Thyroid ...............
Thyroxin ..............
0.5
0.25
1.0
0.02
0.1
0.1
0.005
0.005
0.002115
0.002115
0.0001058
0.0001058
12 = 23.5
12 = 43
12 = 0.17 to 0.23
12 = 64
178

Exercise fi9
Object.-Assay of Spirit of Ethyl nitrite.

Materials Required.-10 cc. of spirit of ethyl nitrite (of known specific
gravity).
Cylinder of carbon dioxide.
10 cc. of dilute hydrochloric acid (1 in 2).
Aldehyde-free alcohol.
Apparatus.-"Fit a 300-cc. Erlenmeyer flask with a 2-hole rubber stopper.
Through one hole pass an aeration tube leading to the bottom of the flask and
tapered to an internal diameter of about 1 mm. at the lower end. Through
the other hole pass a glass tube, of at least 6 mm. internal diameter, which
will extend about 1 cm. above and below the stopper. Connect the aeration
tube with a cylinder or generator of air-free carbon dioxide."
Procedure.-l. "Place 10 Gm. of potassium iodide in the flask and add
40 cc. of boiling, deaerated distilled water. Insert the stopper in the flask
and pass a stream of carbon dioxide through the flask at a rate of 5 bubbles
per second until the solution has cooled to room temperature. Add 10 cc.
of dilute hydrochloric acid (1 in 2) and continue the stream of carbon
dioxide for at least three minutes. If any iodine is liberated, as shown by
the appearance of a yellow color, cautiously adp. tenth-normal sodium
thiosulfate from a burette, through the outlet tube, until the color is just
discharged. "
The water can be deaerated in the flask by boiling it and passing

a stream of carbon dioxide gas through it. Oxygen must be
excluded from the apparatus at all times during the assay since
it would react with the HI formed in the acid solution to liberate
iodine,
If iodine is liberated, the solution becomes slightly yellow in

color indicating that the solution was not completely deaerated.
2. "Decrease the flow of carbon dioxide to about 2 bubbles per second,
and add the directed volume of the nitrite solution with a transfer pipette,
passing the pipette through the outlet tube until the tip is just above the
surface of the potassium iodide solution. Touch the tip of the pipette to
the outlet tube to remove adhering nitrite solution, then rinse the outlet
tube with a fine jet of aldehyde-free alcohol from a wash bottle. At once
titrate the liberated iodine with tenth-normal sodium thiosulfate, introducing
the tip of the burette through the outlet tube."
179
When the sample containing nitrite is introduced into the flask

nitrous acid is form,ed, and it reacts with the HI liberating iodine,
C 2H sONO + HOH~C2HsOH + HN0 2
75.05
2HN0 2 + 2HI--2HOH + 2NO + 12
The liberated iodine is titrated with 0.1 N Na 2S20a in the usual
way.
1. Write equations representing the reactions that occur in the assay of
amyl nitrite and spirit of glyceryl trinitrate.
.
2. Why must air be excluded during the assay of spirit of ethyl nitrite?
3. Ten cubic centimeters of spirit of ethyl nitrite (specific gravity 0.820)
required 30 cc. of a solution of Na 2 S,O., each cubic centimeter of which is
known to be equivalent to. 0.01420 Gm. of iodine. Calculate the per cent
of ethyl nitrite in the sample.
TABLE XXV.-IODOMETRIC NITRITE ASSAYS
Substance
U.S.P.
Amyl nitrite ............
Ethyl nitrite, spirit of ....
Glyceryl trinitratc, spirit
of. .. '................
EquivaAmount lent of
used,
1 cc. of
0.1 N
Gm.
Na,S,O.
per cent
3.5
10
0.01171 C.HuONO = SO
0.007505 C 2H.ONO = 3.5 to 4.5
25
0.0113
C.H.(ONO,).
= 1 to 1.1
TABLE XXVI.-OTHER OFFICIAL SUBSTANCES ASSAYED BY TITRATION OF

THE IODINE LmERATED FROM KI WITH 0.1 N Na,S,O.
Substance
Amount
used,
Gm. or
cc.
U.S.P.
Chromium trioxide ..........
Cupric sulfate ...............
Potassium bromate, R .......
R.
reagent.
1
1
0.3
Equivalent of
1 cc. of
0.1 N
per cent
Na,S 2O.
0.003334 CrO, = 95
0.01596 CUS04 = 63 to 66.S
0.002784 KBrO. = 99.S
180
OXIDATION -REDUCTION METHODS WITH 0.1 N BROMINE
In the assay of compounds such as aniline, phenol, and resorcinol, bromine is employed as an oxidizing agent in place of iodine
since it forms tribrom derivatives quantitatively which are
insoluble in water in the presence of bromine in slight excess.
The volumetric solution does not contain bromine as such, but
it contains potassium bromide and potassium bromate. Mixtures of these salts in solution liberate bromine when treated with
an acid.
Exercise 60
Object.-To Prepare and Standardize 0.1 N Bromine, Koppescharr's Solution.

Materials Required.-3 Gm. of potassium bromate.
25 Gm. of potassium bromide.
5 cc. of potassium iodide T.S. (16.5 Gm. KI in 100 cc.).
Procedure.-Dissolve 3 Gm. of potassium bromate and 25 Gm. of potassium bromide in sufficient distilled water to make 1,000 cc. and determine its
exact normality as follows:
"Measure accurately from a burette about 25 cc. of the'solution into a
500-ee. glass-stoppered flask and dilute with 120 ee. of distilled water. Add
5 ec. of hydrochloric acid, stopper the flask, and shake it gently. Then add
5 cc. of potassium iodide T.S., re-stopper, shake the mixture, allow it to
stand for five minutes, and titrate the liberated iodine with tenth-normal
sodium thiosulfate, using starch T.S. as the indicator. Calculate the
normality and, if desired, adjust exactly to tenth-normal."
When the solution of potassium bromide and potassium bromate is acidulated with hydrochloric acid, bromine is set free:
5KBr
+ KBrOa + 6HCl~6KCl + 3Br2 + 3H20
and the liberated bromine replaces the iodine of KI:

2KI
+ Br2~2KBr + 12
The flask should be tightly stoppered after the addition of the

acid to prevent the escape of bromine vapors.
The free iodine is titrated with 0.1 N sodium thiosulfate to
standardize the solution:
12
+ 2Na2S208~2Nal + Na22406
181
Since 1 mole of N a2S20a is equivalent to 1 Gm.-atom of iodine,

and 1 Gm.-atom of iodine is equivalent to 1 Gm.-atom of bromine, each cubic centimeter of 0.1 N Na2S20a is equivalent to
10 1~6i~~00 = 0.012692 Gm. iodine and to 10 ~'i~oo = 0.007992
Gm. bromine.
If 20 cc. of 0.1 N bromine require 25 cc. of 0.1 N sodium thiosulfate, the solution would be 25
~o 0.1 =
0.1250 normal.
Each
20 cc. of the bromine solution may be diluted to 25 cc. to make

an exactly 0.1 N solution, or the solution of known normality may
be employed.
Exercise 61
Object.-Assay of Phenol.
Materials Required.-1.5 Gm. of phenol.
30 cc. of 0.1 N bromine solution.
1 cc. of chloroform.
Procedure.-l. Dissolve about 1.5 Gm. of phenol, accurately weighed,
in sufficient distilled water to make 1,000 cc. Transfer an aliquot portion of
this solution, containing from 0.038 to 0.041 Gm. of phenol, to a 500 cc.
glass-stoppered flask having a long, narrow neck, add 30 cc. of 0.1 N bromine, then 5 cc. of hydrochloric acid', and immediately insert the stopper.
Shake the flask repeatedly during :hi hr., allow it to stand for 15 min.,
remove the stopper just sufficiently to introduce quickly 5 ce. of an aqueous
solution of potassium iodide (1 in 5), being careful that no bromine vapor
escapes, and at once stopper the flask.
When the HCI is added, bromine is set free:

5KBr
+ KBrOa + 6HCI~6KCl + 3Br2 + 3H 0.

2
The bromine reacts with phenol to form a white crystalline

precipitate of tribromphenol and hydrobromic acid:
CsH.OH
94.08
+ 3Br2~CsH2Br30H + 3HBr
The flask should be tightly stoppered to prevent. the escape of the

volatile bromine and shaken gently during 30 min. to mix the
182
contents thoroughly and allow the 6xidation of phenol to tribromphenol to go to completion.

2. Shake the flask thoroughly, remove the stopper, and rinse it and the
neck of the flask with a little distilled water, so that the washings may flow
into the flask, then add 1 cc. of chloroform, shake the mixture well, and
titrate with 0.1 N sodium thiosulfate, using starch T.S. as indicator. Each
cubic centimeter of 0.1 N bromine corresponds to 0.001568 Gm. of C 6H.OH.
After the addition of potassium iodide, 1 cc. of chloroform is

added to dissolve the precipitated tribromphenol which -would
otherwise interfere with the clear observation of the end point,
particularly in the assay of old, colored phenol. The excess
bromine liberates iodine in accordance with the following
equation:
The free iodine reacts with sodium thiosulfate:

12 +
Na2S203~2Na1
+.Na2S406
The number of cubic centimeters of 0.1 N bromine added less

the number of cubic centimeters of 0.1 N Na 2S2Oa required in the
titration gives the number of cubic centimeters of 0.1 N bromine
consumed in the reaction with phenol.
I
Since each mole of phenol oxidized to tribromphenol requires
6 Gm.-atoms of bromine, each cubic centimeter of 0.1 N bromine
94.08
consumed is 'equivalent to 6 X 10 X 1,000 = 0.001568 Gm.
C 6H 50H.
Liquefied phenol and resorcinol are assayed by the same
method. Acetanilid in tablets of acetanilid is determined by a
special method which is based upon the formation of tribromaniline during direct t~tration with a standard bromine solution.
1. What substance is the oxidizing agent in Koppescharr's solution1

2. Explain the derivation of the equivalent weight of KBrO. when it is
used as an oxidizing agent in accordance with the change of charge which
the bromine atom undergoes.
3. How much pure KBr and pure KBrO. must be present in a solution
which when treated with an acid will liberate an amount of bromine equivalent to 100 cc. of 0.1 N iodine solution?
183
4. Calculate the weight of phenol and resorcinol, respectively, equivalent

to 25 cc. of 0.1220 N bromine?
5. Aniline forms a tribromaniline when treated with bromine analogous
to tribromphenol. Write equations for the reactions that occur in the assay
of acetanilid in tablets of acetanilid.
6. Upon standardizing a solution of potassium bromide-bromate against
acetanjlid (see assay.of tablets of acetanilid), 1 cc. of the solution was found
to be equivalent to 0:008240 Gm. of acetanilid. A 0.2856 Gm. sample of
powdered acetanilid tablets required 21.30 cc. of the bromide-bromate solution. If 20 tablets weighed 6.1244 Gm., calculate the average amount of
acetanilid per tablet.
Exercise 62
Object.-Assay of ammonium hypophosphite.

Materials Required.-O.15 Gm. of ammoni~m hypophosphite.
oj N bromine.
Procedure.-" Accurately weigh about 0.12 Gm. of the salt, dried over
sulfuric acid for 24 hours, and dissolve it in sufficient water to make 100 cc.
Transfer 50 cc. of the solution to a 250-cc. glass-stoppered flask, add 50 cc.
of tenth-normal bromine solution and 20 cc. of diluted sulfuric acid, stopper
the flask, shake well and allow to stand for three hours. Add 2 Gm. of
potassium iodide, dissolved in 10 cc. of recently boiled distilled water,
shake the flask, and titrate the liberated iodine with tenth-normal sodium
thiosulfate, using starch T.S. as the indicator. Each cubic centimeter of
tenth-normal bromine is equivalent to 0.002077 Gm. of NH,PH 20 2."
The hypophosphorous acid, formed when H 2S0 4 is added to

the solution of NH 4PH 20 2, is oxidized quantitatively to phosphoric acid upon standing in contact with the bromine solution:
2NH~H202
+ H2S04~(NH4hS04 + 2HPH 20 2
2(83.07)
H
/
HO-P=O
'"
HO
'"
+ 2Br2 + 2H20~HO-P
66.04
4(79.916)
+ 4HBr
HO/
98.04
Two of the hydrogen atoms in HPH 20 2 carry negative charges

and are oxidized by the Br2 to positive hydrogen. The phosphorus undergoes no change of charge. The ionic equation
representing the reaction that occurs may be written:
184
2H- + 2Br2~2H+ + 4Bc

2( -) + zero = 2( +) + 4( - )
From the reactions, it is apparent that each cubic centimeter of
0.1 N bromine consumed is equivalent to 4 X

0.002077 Gm. of NH 4PH 20 2
18;~71,000

1. How is ferric hypophosphite assayed? (See N.F. VI.)
2. Derive the factor for total hypophosphites equivalent to 1 cc. of
0.1 N bromine for a mixture containing 50 Gm. of Ca(PH20 2)2, 20 Gm. of
NaPH 20 2, and 30 Gm. of KPH202 assuming 100 per cent purity for each
of the salts.
3. Give examples of a number of official substances other than those listed
in the appended table to which this method of assay might be applied.
TABLE XXVII.-OFFICIAL SUBSTANCES ASSAYED BY TITRATING WITH
0.1 N
SODIUM THIOSULFATE THE IODINE LIBERATED FROM POTASSIUM IODIDE

BY ExCESS
Substance
U.S.P.
Phenol .......... '" ..
Phenol, liquefied .......
Phenol ointment .......
Resorcinol. ...........
Amount
used,
grams
0.1 N
BROMINE
Equivalent of
1 CC.
0.1 N
bromine
1.5
1.5
2
1.5
0.001568
0.001568
0.001568
0.001834
0.12
0.12
0.002077 NH.PH 20. = 97.5

0.002127 Ca(PH20 2). = 98
0.12
0.002538 Mn(PH 20 2)..H.O = 97
0.12
0.002604 KPH20. = 98
0.001834 C6H.(OH). = 9.5 to 10.5
1
0.12
0.001834 C.H.(OH)z = 19 to 21
0.002651 NaPH.O . H 20 = 98
0.002668 C 6H . OH.COONa
C6H sOH =
C6H,OH =
C.H,OH =
C.H.(OH)2
98
88
1.8 to 2.2
= 99.5
N.F.
Ammonium hypophosphite ...............

Calcium hypophosphite
Manganese hypophosphite ...............
Potassium hypophosphite .. " ...........
Resorcinol, mild paste
of. ................
Resorcinol, strong paste
of .................
Sodium hypophosphite
Sodium salicylate in
caffeine with sodium
salicylate ...........
= 48 to 52
185
OXIDATION-REDUCTION METHODS WITH POTASSIUM IODATE
Potassium iodate may be used as the oxidizing agent in the

assay of a number of substances such .as iodides, arsenites, and
other reducing agents. The method depends upon the formation
of iodine mono chloride in strong hydrochloric acid solution.
Exercise 63
Object.-To Prepare 200 cc. of 0.05 Molar Potassium Iodate.

Materials Required.-About 2.2 Gm. of potassium iodate.
Procedure.-Dry 2.2 Gm. of potassium iodate to constant weight at
110C. Dissolve 2.1402 Grn. of the dried salt in sufficient distilled water to
make exactly 200 cc. at 25C. The resulting solution should be exactly
0.05 molar.
The potassium iodate is dried to constant weight at 110C.

to insure its freedom from moisture. Since potassium iodate is a
very stable salt and can be obtained in a very pure condition,
standard solutions of this reagent can be prepared by dissolving
the calculated weight of the salt in distilled water and diluting
to the proper volume.
Standard iodate solutions of known molarity rather than those
of known normality are usually employed because the normality
varies depending on the nature of the reaction, e.g.,
1. Potassium iodate reacts with iodine in the presence of 12 per
cent or more of HCI as follows:
and a normal solution of KIO a with respect to iodine should

contain }i mole.
2. Potassium iodate reacts with KI in the presence of 12 per
cent HOI as follows:
2KI
+ K10 + 6HCI~3KCI + 31CI + 3H~O

3
and a normal solution of KIO a with respect to KI should contain

72 mole.
3. Potassium iodate reacts with As 20 a in acid solution as
follows:
186
and a normal solution of KIO a with respect to As 20 a should

contain X mole.
Exercise 64
Object.-Assay of Potassium Iodide.

Materials Required.-O.5 Gm. potassium iodide.
0.05 M potassium iodate.
Procedure.-" Dry about 0.5 Gm. of Potassium Iodide to constant weight
at 100C., weigh accurately, dissolve it in about 10 cc. of distilled water, add
35 cc. of hydrochloric acid and 5 cc. of chloroform. Titrate the mixture with
twentieth-molar potassium iodate until the purple color of iodine disappears
from the chloroform. The last portions of the iodate solution must be
added in drops, the mixture being agitated vigorously and continuously.
After the chloroform has been decolorized, allow the mixture to stand for five
minutes. If the chloroform develops a purple color, the mixture should be
titrated further with the iodate solution. Each cubic centimeter of twentieth-molar potassium iodate is equivalent to ,0.01660 Gm. of KI."
Potassium iodate is quantitatively reduced to iodine chloride

in strong HCI solution. Iodine is formed intermediately but at
the equivalent point, in the presence of sufficient :trCI, it is completely converted into ICI. The titration mixtufe should contain
at least 12 per cent of HCI at the end point to preve~t th~ hydrolysis of ICI. In solutions slightly acidified with HCI, the equation
representing the reaction is:
5KI
+ KIO a + 6HCI--).6KCI + 31 2 + 3H 20
When sufficient HCI is present in the reaction mixture, the equation representing the reaction is:
2K1 + KIO a
2(166.02)
+ 6HCl-3KCI + 31Cl + 3H 2 0
The change of charge in the latter equation is as follows:

21-
+ 15+-31+
2(-)+5(+) =3(+)
Each atom of iodine combined as iodide loses 2 electrons and each
atom of iodine combined as iodate loses 4 electrons. Each cc. of
0.05 M potassium iodate is equivalent, therefore, to 2~(~6i~~~0
0.01660 Gm. of KI.
187
Chloroform is added to the titration mixture to enable the easy

and accurate determination of the end point. Since all of the
iodine which remains during the latter part of the titration is
collected in the chloroform layer, the disappearance of the violet
iodine color is readily observable. Vigorous shaking of the
titration mixture ifl, necessary as the end point of titration is
approached to mix intimately the immiscible solvents so that the
iodine in the chloroform layer will be brought in contact with the
iodate.
1. Why should the titration of iodides with iodate be carried out in
strongly acid solution?
2. What is the function of the chloroform in the assay of iodides with
potassium iodate?
3. Write equations representing the change of charge that occurs when
iodate is added to a solution of iodide which is slightly acid.
4_. 40 tablets of red mercuric iodide labeled" 715 grain tablets of mercuric
iodide" weighed 12.4220 Gm. A 10.0000 Gm. sample of the powdered
tablets required 14.45 cc. of 0.02 M KIO.. Calculate the amount of HgI 2
per tablet. Calculate the per cent deviation from the labeled amount.
6. Ascertain the method of assay of iodine ointment in the U.S.P. Explain the procedure.
TABLE XXVIII.-O~'FICIAL SUBSTANCES ASSAYED BY TITRATION WITH
Amount
Substance
used,
Om. or
cc.
Equiva-
Molarity
lent of
of KIO.
1 cc.
KIO
per cent
--U.S.P.
Chiniofon, powder . . ............
Iodophthalein, soluble ..........

Oil, iodized ....................
Potassium iodide ............. , .
Sodium iodide .................
0,3
0.3
0.35
0.5
0.5
0.05
0.05
0.05
0.05
0.05
0.060.2
0.13-
0.02
0.05
",0.02
O.la
0.02
0.05
0.02
0.05
0,01269
0.01269
0.01269
0.01660
0.01499
I = 26.5 to 28.9
I = 61 to 62
I = 39 to 41
KI = 99
NaI = 99
N.F.
Arsenic trioxide, tablets of .......

Mercuric iodide, red . ...........
Mercuric iodide, r-ed, tablets of ..

Mercurous iodide, yellow, tablets
of. ........................
Potassium iodide, solution of ....
Potassium iodide, tablets of .... , .
Sodium iodide, ampuls of. .......
- Amount of ingredient sought.
5
O.la
0.5"
0.003956 As,O. = 92.5 to 107.5'

0.02272 HgI, - 99
0.009089 HgI, = 91 to 109'
0.008734
0.01660
0.006640
0.01499
HgI = 91 to 109'
KI = 97 to 103W/Y
KI = 92.5 to 107.5'
NaI = 92 to 105'
CHAPTER X
GASOMETRIC METHODS
The assay processes which involve the volumetric measurement of gases are of two types, namely:
1. The purity of a gas is determined by absorbing one of the
components of a gaseous mixture by means of a suitable absorbing
agent and measuring the quantity of gas absorbed by the change
in volume.
2. The purity of a given substance is determined by measuring
the quantity of gas which it may be made to evolve in a given
chemical reaction.
Theory.-According to the kinetic theory of gases, the molecules of a gas are in constant motion. When a gas is enclosed in a
vessel, it exerts a definite pressure due to the combined effects of
the impacts of the moving molecules of gas upon the walls of
the containing vessel. The magnitude of the pressurE\ exerted
depends on the number of molecules which strike a given surface
in a given period of time. The number of molecules which strike
upon the surface in a given period of time is dependent upon the
number of molecules present and upon the velocity of the molecules. The velocity of the molecules varies with the temperature
of the gas; the higher the temperature the greater the velocity of
the molecules, and the lower the temperature the less the velocity
of the molecules. If the temperature and pressure of a gas remain
constant, the volume of the gas will depend on the number of
molecules present.
The official gasometric determinations pertain only to substances which behave as "perfect" gases, that is, gases in which
there is no dissociation or association of the molecules, and which,
therefore, obey the gas laws. Consequently, a definite volume
of such a gas measured under laboratory conditions at a known
temperature and pressure provides the data required to calculate
the weight or volume of the gas present under standard condi188
GASOMETRIC METHODS
189
tions, 'as will be shown subsequently. The term standard conditions is used to designa:te a temperature of 00. and a pressure
equal to that exerted by a column of mercury 760 mm. in height.
Whenever the volume of a gas is given, it is understood that the
gas occupies the specific volume when measured at 00. and
at a barometric pressure of 760 mm. unless otherwise stated.
Standard conditions should not be confused with the expression
normal conditions which has reference to the measurement of
gases at 250. and 760 mm. pressure.
It has been found by experiment that 22.4: l. of oxygen measured at 00. under a pressure of 760 mm. weighs 32 Gm. and that
the same volume of hydrogen measured under standard conditions
weighs 2.016 Gm. These facts are embodied in Avogadro's
hypothesis, which is: Equal volumes of gases measured under the
same conditions of temperature and pressure contain the same
number of molecules. In the case of perfect gases, this hypothesis
may be stated: Equivalent molecular weights of gases confined
under identical conditions of temperature and pressure occupy the
same volume. Thus, 2.016 Gm. of hydrogen, 32 Gm. of oxygen,
30.01 Gm. of nitric oxide, or 14:.008 Gm. of nitrogen will occupy
22.4: l. when measured at 00. and 760 mm. pressure. According
to Boyle's law: The volume of a gas is inversely proportional to
the pressure upon it if the temperature remains constant. Doubling
of the pressure would diminish the volume one-half, and, conversely, diminishing the pressure one-half would double the
volume. Th,e product of the pressure and volume is therefore
constant. Boyle's law may be stated in algebraic form as follows:
or
VI = P 2 = K
V 2 PI
where PI and VI are the pressure and vollfme under one set of
conditions and P 2 and V 2 are the pressure and volume under
another set of conditions, K being their constant product. The
magnitude of K depends upon the quantity of gas measured. If
PI and VI are used to represent the observed pressure and volume
of a gas, respectively, and P 2 is standard pressure (760 mm.),
the volume V 2 which the gas will occupy at P 2 may be calculated;
e.g., if a sample of gas measured at 25C. and 750 mm. pressure
occupies a volume of 50 cc., what volume will it occupy at 25C.
190
and 760 mm. press11re? By makirlg the proper substitutions

in the preceding equation,
50
760
VI = 750
and
VI =
50 X 750
760
= 49.35 cc.
The increase of 10 mm. pressure on 50 cc. of gas measured at

25C. is thus found to have caused a decrease in volume of 0.65 cc.
Charles' law may be expressed as follows: If the volume of a
definite mass of gas is kept constant, the pressure which it exerts
is directly proportional to the absolute temperature at which it is
measured. This law may be stated in algebraic form as follows:
or
where VIand V 2 are the volumes of the gas at the absolute temperatures Tl and T 2 Zero on the absolute temperature scale
corresponds to -273C. The absolute temperature at which
a given analysis is carried out is therefore found by adding 273
to the laboratory temperature expressed in degrees centigrade.
The above equation then becomes
VI
273
V 2 = 273
+i
+ t2
and
V _ V 2 X 273
I 273
t2
tl
or I
where tl and t2 represent the respective temperatures in degrees

centigrade; e.g., if a sample of gas measures 50 cc. at 25C.,
what volume would it occupy at ODC., the pressure remaining
constant? Upon substituting in the above equation, we obtain
.50
V2
273 + 25
273 + 0
and
V2
_ 50 X 273
298
= 45.81 cc.
When both the temperature and pressure at which a gas is

measured deviate from. the official requirement, the corrections
for each may be made in succession, as illustrated in the above
examples. It is more convenient, however, to combine the
respective equations which represent Boyle's law and Charles'
law and make the correction in a single calculation. The combined laws may be expressed by the algebraic equation
V - P 2V 2T 1
1 -
P 1T 2
GASOMETRIC METHODS
191
The following example illustrates the use of this equation. A gas

measures 40 cc. at 200. and 740 mm. pressure. What volume
will it occupy at 250. and 760 mm. pressure? By substituting
the proper values in the foregoing equation, we obtain
740 X 40 X 298
V1 =
760 X 293
= 39.75 cc.
Apparatus.-The volumetric measurement of gases in the
official assay processes is made by the use of a gas buretpe of the
FIG. 15.-Gas
burette with level
ling flask. (Cour.
tesy Fisher Scienti
fie Company.)
FIG. 16.-Simple gas pipette.

(Courtesy Fisher Scientific Company.)
type illustrated in Fig. 15. The burette should be of 100 cc.

capacity and graduated in 0.2 cc. divisions. The burette may
be enclosed in a water jacket, "as illustrated, to permit the
control of temperature. The lower end of the gas burette is
attached to an open leveling or equilibrium tube by means of a
heavy walled rubber tube.
The absorption of the gases is accomplished in a gas pipette
similar to that shown in Fig. 16. The absorption bulbI of the
192
pipette is provided with an opening at the bottom which can be

closed by means of a rubber stopper and through which solid
absorbing agents may be introduced.
Exercise 65
Object.-Assay of Carbon Dioxide.

Materials Required.-Carbon dioxide compressed in a metallic cylinder
which is provided with a reducing valve.
A gas burette with leveling tube.
A gas pipette.
About 2 lb. of mercury.
150 cc. of 50 per cent KOH solution.
Procedure.-l. "Place a sufficient quantity of mercury in a 100-cc. gas
burette or nitrometer, provided with a two-way stop-cock and a two-way
outlet, and properly connected with a balancing tube. Connect one of the
intake tubes of the nitrometer with a gas pipette of suitable capacity.
Place in the pipette about 125 cc. of 50 per cent potassium hydroxide solution. Draw the liquid (free from air bubbles) through the capillary opening,
connection and stop-cock opening in the nitrometer by reducing the pressure
in the nitrometer tube and opening the stop-cock controlling the connection
with the gas pipette. Then close the stop-cock."
The gas burette is filled with mercury by raising the leveling

flask containing the mercury until the stopcock opening is filled
with the mercury. The stopcock is then closed.' The gas
pipette is filled with 50 per cent KOH so that the absorption bulb
and capillary tube are completely full and the expansion bulb
is about one-tenth full and is connected to one of the outlet tubes
of the gas burette. Upon opening the stopcock connecting the
pipette and the burette and lowering the leveling flask, the KOR
solution and any trapped air in the connection are drawn through
the stopcock opening. Any air bubbles drawn into the burette
may be driven out by opening the stopcock to the other outlet
and raising the leveling flask.
2. "Having completely filled the nitrometer, the other stop-cock opening
and the other intake tube with mercury, draw into the nitrometer exactly
100 cc. of Carbon Dioxide, measured at atmospheric pressure and at 25C.,
by reducing the pressure in the tube."
The gas can be introduced directly into the burette from a

cylinder of the compressed gas if the latter is provided with a
reducing valve which can be carefully controlled. When the
193
GASOMETRIC METHODS
gas is obtained in this manner, the connection tubing between

the cylinder and the burette should be flushed free of air by
allowing a stream of the gas to flow through the tubing for a
minute or two. A volume of gas slightly in excess of 100 cc.
is then admitted to the burette, and after it has acquired a
temperature of 25Q., the excess gas is permitted to escape so
that a volume of 100 cc. remains.
3. "Close this stop-cock. Increase the pressure on the Carbon Dioxide
in the nitrometer tube and open the stop-cock controlling the connection
with the gas pipette. Force the entire volume of gas into the pipette.
Close the stop-cock and rock the pipette gently, providing frequent contact
of the liquid and gas. At the end of five minutes most of the gas will have
been absorbed by the liquid. At this time, to facilitate the absorption of
the last portion of the gas, draw some of the liquid into the nitrometer tube
and force the residual gas back upon the surface of the liquid in the gas
pipette. Again rock the pipette until no further diminution in the volume
of gas occurs. Draw the residual gas, if any, into the nitrometer tube and
me'asure its volume at atmospheric pressure and at 25C.
"The volume of gas remaining undissolved should not exceed 1 cc., indicating not less than 99 per cent by volume of CO 2 i~ the gas tested."
The CO 2 is absorbed by the KOR forming K 2 CO a leaving other

gases which are not absorbed by KOR as residual gas. If the
initial volume of the sample was exactly 100 cc., the decrease
in volume is numerically equal to the per cent of CO 2 in the
sample.
All the official gasometric assays are performed by the absorption method. The procedures are designed primarily for use in
manufacturing laboratories for the control of the purity at the
time the gases are compressed into cylinders.
TABLE XXIX.-OFFICIAL GASOMETRIC ASSAYS
Substance
U.S.P.
Carbon dioxide .. ....
Ethylene ............
Nitrogen monoxide ..
Oxygen .............
Absorption reagent
Potassium hydroxide
Bromine
Water
Copper with NH.CI and
NH 40H
CO. = 99
CH. = CH 2 = 99
N 20 = 95
0=99
194
Although no assay process involving the measurement of

evolved gases is official, the procedure employed in this type
of assay is of such general application that every student of
pharmaceutical analysis should be familiar with the method.
The following exercise illustrates this method
of gasometric analysis.
Apparatus.-The volumetric measurement of
evolved gases may be made by the use of an
instrument known as a nitrometer. The nitrometer of the type devised by Lunge (Fig. 17)
consists of a gas burette A of at least 50 cc.
E
capacity, graduated into 0.1 cc., with a special
A
glass stopcock B at the top, to which is. sealed a
glass cup D. The gas burette is connected with
an open leveling or equilibrium tube E by means
of a heavy-walled rubber tube F.
In the use of the simple Lunge apparatus
described above, both tubes of the instrument are
_;;;;::j..........c-F
partly filled with some liquid in which the gas to
be measured is insoluble; generally mercury is
FIG. 17.-Lunge employed but in some cases a saturated salt
nitrometer.
solution is used. The latter may \be used in the
measurement of the nitric oxide liberated from ethyl nitrite or
amyl nitrite.
When mercury is used, enough of it is poured irtto the equilibrium tube to fill the burette and leave a slight excess. The
stopcock is turned to connect the gas burette with the glass cup,
and the equilibrium tube is raised until the mercury level reaches
the bottom of the cup to expel all of the air from the burette
and stopcock bore, and the stopcock is closed. The sample to
be assayed, a liquid or a solid in solution, is weighed and poured
into the cup from a weighing bottle or weight pipette. Upon
opening the stopcock and lowering the equilibrium tube slightly,
the sample is drawn into the burette. Care must be exercised ~o
prevent the entrance of air into the passage of the stopcock. The
cup is rinsed two or three times with 3 cc. portions of the reagents
used to liberate the gas, each being drawn into the burette as
described above. With the stopcock closed, the equilibrium
tube is lowered sufficiently to cause a slight diminution of the
GASOMETRIC METHODS
195
pressure in the burette, and the latter is shaken vigorously for

about 5 min. to bring the reagents and sample into intimate
contact. When no' more gas is evolved on further shaking, the
burette is allowed to stand about 30 min. to permit its contents
to acquire room temperature. The volume is then measured at
atmospheric pressure. Since there is a column of impure reaction mixture above the mercury in the burette, some procedure
must be employed to adjust the level of the equilibrium tube so
that the weight of the column of mercury in it will just equal the
weight of the column of mercury and reaction mixture in the
burette. If the reagent mixture has a specific gravity of about
1.3, as it does in the assay for ethyl nitrite, the equilibrium
tube is set on the assumption that 1 mm. of mercury will balance
about 10 mm. of the reaction mixture. About 5 cc. of dilute
sulfuric acid is then poured into the cup and the stopcock is
carefully opened to it. The tendency to draw in or expel gas is
noted and the equilibrium tube is adjusted accordingly. This
tedious leveling procedure is obviated in the assay by the use of
a saturated solution of sodium chloride in the place of mercury.
Exercise 66
Object.-To Prepare and Test a Nitrometer.

Materials Required.-l gas burette.
1 equilibrium tube.
30 in. of nitrometer tubing.
A saturated solution of sodium chloride.
Procedure.-Clean the gas burette and equilibrium tube of the nitrometer
thoroughly, connect them by a piece of heavy-walled rubber tubing about
30 in. long, grease the burette stopcock with a thin film of petrolatum, and
clamp the assembled instrument securely to a support in a vertical position.
Open the stopcock of the burette and pour sufficient saturated solution of
sodium chloride into the equilibrium tube to fill the burette and the bore of
the stopcock completely when the equilibrium tube is raised. Carefully
adjust the level of the equilibrium tube so that the stopcock passage is
completely filled with the salt solution without any of it being present in the
glass cup and close the stopcock. Lower the equilibrium tube so that the
solution level in it corresponds approximately with the 50 cc. graduation
on the burette, note the solution level in the burette, and after the instrument
has stood for 20 min. take another reading. If the two readings do not
correspond, the stopcock is leaky. If the leakage cannot be corrected by
the use of stopcock grease, another instrument should be employed.
196

Exercise 67
Object.-Assay of Spirit of Ethyl Nitrite.

Materials Required.-1 Lunge nitrometer.
1 10 cc. delivery pipette.
1 100 cc. volumetric flask.
40 cc. of spirit of ethyl nitrite.
0.5 Gm. of potassium bicarbonate.
60 cc. of alcohol.
1.65 Gm. of potassium iodide.
Procedure.-1. Transfer about 40 cc. of spirit of ethyl nitrite, which
has been previously shaken with 0.5 Gm. of powdered potassium bicarbonate, to a tared, 100 cc. measuring flask, and weigh accurately. Add sufficient alcohol to bring the volume to exactly 100 cc. and mix thoroughly.
In the presence of water, ethyl nitrite is partially hydrolyzed

with the formation of ethyl alcohol and nitrous acid:
C 2H 5N0 2
+ H20~C2H50H + HN0 2
Since the nitrous acid, if present, would liberate oxide in the

assay, it is removed by shaking the spirit with powdered potassium bicarbonate:
HN0 2 +
KHC03~ KN0 2
+ H 20 + <\)0 2
The potassium nitrite formed is practically insoluble in the spirit.

The sample is diluted with alcohol to 100 cc. after weighing, so
that an aliquot portion of the solution representing a definite
weight of the spirit may be measured into the nitrometer because
it is difficult to weigh and transfer a sample of the spirit directly
without the loss of some ethyl nitrite.
2. Introduce into a nitrometer exactly 10 cc. of the alcoholic solution,
follow by 10 cc. of potassium iodidll T.S. and afterwards by 5 cc. of diluted
sulfuric acid. When.the volume of gas has become constant (within 30 to
60 min.) note the volume of gas ,collected.
Adjust the nitrometer prepared for use in the preceding

exercise so that the level of the salt solution in the equilibrium
tube is about even with the 15 cc. graduation mark on the burette,
the latter as well as the stopcock passage being filled with the
salt solution. Fill a 10 cc. pipette with the diluted spirit exactly
to the graduation mark and transfer the liquid into the nitrometer
GASOMETRIC METHODS
197
cup. Open the stopcock at once to allow the solution to be drawn

into the burette using care to prevent the entrance of air at the
same time. Rinse the cup at once with two 5 cc. portions of
potassium iodide solution (1.65 Gm. KI in 10 cc. H 2 0), drawing
each portion into the burette by lowering the equilibrium tube,
if necessary, and then place about 6 cc. of diluted sulfuric aGid
in the cup and draw about 5 cc. of it into the burette in the same
manner. The excess acid remaining in the cup serves to prevent
the escape of the nitric oxide gas which begins to form as soon as
the first of the acid is introduced, and it prevents the entrance of
air. After the stopcock has been tightly closed and the equilibrium tube well clamped to the support, remove the burette
from the clamp, incline it at an angle of about 45 deg., and shake
it vigorously for about 5 min. to mix the reagents thoroughly
with the solution of ethyl nitrite.
The sulfuric acid liberates hydriodic acid from the potassium
iodide:
.
KI
+ H2S0r--~KHS04 + HI
and the liberated hydriodic acid reacts with the ethyl nitrite in
the spirit, setting nitric oxide gas free:
20 2H 5N0 2 +
2HI~202H60H
+ 12 + 2NO
The entire reaction may be represented in one equation, as:
2KI
+ 202H
N02 + H2S04~202H50H
+ 12 + 2NO + K 2S0 4
The nitric oxide gas formed being insoluble in saturated salt

solution rises to the top of the burette, and after it has acquired
room temperature as shown by the constancy of its volume when
two successive readings taken at 10-min. jnterv~ls agree, the
volume of gas liberated is recorded. When the final reading of
the volume of gas is taken, the liquids in the burette and equilibrium tube should be on the s~me level.
Since 1 Gm.-molecular weight of a gas at 00. and 760 mm.
pressure occupies 22.4 l., 1 l. of a gas will contain the grammolecular weight divided by 22.4. The volume of 1 Gm.-molecul~r weight (30.01Gm.) of nitric oxide gas measured under 00.
and 760 mm. pressure, is 22.3855 l. One liter of nitric oxide gas
will therefore contain 30.01/22.3855 = 1.3406 Gm., and 1 cc.
198
QUANTITATIVE PHARMACEUTICAL CHEMISTRY,
of the gas will contain 0.0013406 Gm.
At 25C. and 760 mm.
pressure, 1 l. of the gas will contain 1.3406 X
2732~ 25
1.2281 Gm., and each cubic centimeter of nitric oxide gas will
contain 0.0012281 Gm. of NO.
The volume of gas being known and the temperature in degrees
centigrade and the barometric pressure in the laboratory having
been ascertained, the weight of nitric oxide gas evolved by a
given weight of spirit of ethyl nitrite may be calculated; e.g., an
aliquot portion of an alcoholic solution of spirit of ethyl nitrite
equivalent to 4.0 Gm. of the spirit when assayed evolved 50 cc.
of nitric oxide measured at 21C. and 740 mm. pressure. What
percentage by weight of ethyl nitrite did the sample contain?
Substituting in the algebraic equation which expresses the combined laws of Boyle and Charles,
2V 2 T t
we obtal'n V = 740 X 50 X 298 = 4935
V t = PPlTl
1
760 X 294
. cc.,
the volume corrected to 25C. and 760 mm. pressure. 49.35 X
0.0012281 = 0.06061 Gm. NO contained in 49.35 cc., at 25C.
and 760 mm. pressure.
~
Since 1 mole, 75.05 Gm., of ethyl nitrite, C 2H oN0 2, evolves 1
mole, 30.01 Gm., of nitric oxide, NO, each gram nitric oxide is
the approximate equivalent of 75/30 = 2.5 Gm. ethyl nitrite,
and 0.06061 Gm. of NO is equivalent to 0.06061 X 2.5 = 0.1515
Gm. C 2H oN0 2
The percentage by weight of ethyl nitrite contained in the
.
0.1515 X 100
4.0
= 3.79 per cent. Calculate
sample IS therefore
0,
the percentage of ethyl nitrite present in the sample of spirit

assayed.
Conunent.-When a gas is measured above a liquid, the total
pressure observed'is due to the vapor pressure of the liquid plus
the pressure of the gas. In the assay of spirit of ethyl nitrite,
the nitric oxide gas evolved is measured above a solution' mixture
consisting primarily of water and alcohol. According to Dalton's
law, each gas in a mixture of gases exerts the same pressure as it
would if it were present alone. The vapor pressure of water
at 25C. has been found to be equal to 23.55 mm. of mercury at
DoC., and that of alcohol has been found to be equal to 5.94 mm.
of mercury at OC. The pressure exerted by the nitric oxide gas
GASOMETRIC METHODS
199
in the preceding example should, therefore, be corrected for the

vapor pressure of water and alcohol as follows: 740 - (23.55
+ 5.94) = 710.5 mm. Unless this correction is made, the method
of assay measures the water and alcohol vapors present as nitric
oxide.
1. Upon what factors is the pressure exerted by a gas Citlpendent?

2. How do the "normal conditions" employed in the measurement of
gases in the Pharmacopoeia differ from standard conditions?
3. How many grams of oxygen, hydrogen, and nitric oxide gas, respectively, will be contained in a volume of 1 l. measured at OC. and 760 mm.
pressure"?
4. 60 cc. of nitric oxide gas is cooled at constant pressure from 40 to 25C.
Calculate the volume of gas at the latter temperature.
6. A gas occupies a volume of 40 cc. at 20C. and 765 mm. pressure.
What volume will it occupy undBr standard conditions?
.
6. An aliquot portion equivalent to 4.2 Gm. of spirit of ethyl nitrite
evolves 48 cc. of nitric oxide measured at 25C. and 742 mm. pressure.
Calculate the percentage by weight of ethyl nitrite contained in the spirit.
7. If the gas in problem 6 was collected over a solution consisting primarily
of alcohol and water with vapor pressures equal to 5.94 mm. and 23.55 mm.
of mercury, respectively, what percentage of ethyl nitrite would the solution
contain?
8. From the results obtained in problems 6 and 7 calCUlate the per cent
of error in the a,mount of ethyl nitrite contained in the spirit that is caused
when the corrections for the vapor pressures of water and alcohol are not
made.
9. Write equations representing the reactions that take place in the
absorption of the gases in the official gasometric assays.
PART II
PHYSICO-CHEMICAL METHODS USED IN
'OFFICIAL PHARMACEUTICAL ANALYSIS
The anaiysis of many of the official substances for the purpose
of identifying them and of determining their purity or therapeutic value is based on the measurement of certain physical and
chemical characteristics or constants, considered in conjunction
with the estimation of certain special components by assay, and
qualitative tests for various substances commonly present as
i:rb.purities or added for the purpose of adulteration.
The qualitative methods employed in the determination of the
physical characteristics or physical constants of official substances
are frequently applied to materials of widely divergent nature;
e.g., valuable criteria relative to the purity of volatile oils, sugars,
and alkaloids may be gained by the measurement of the optical
activity. The fundamental principles involved in ascertaining
the physical constants of these substances are the same regardless
of the nature of the substance, although slight modifications
of the technique employed are frequently necessary.
The Pharmacopoeia and National Formulary, as a rule, specify
that certain methods and apparatus shall be employed in the
determination of the physical constants of official substances.
In such cases, the methods are known as official methods and the
apparatus as official apparatus.
CHAPTER XI
SOLUBILITY
The statements of the solubilities of official substances are
given primarily jor the information of physicians and pharmacists
for use in the preparation and dispensing of medicines. They
should not always be regarded as exact physical constants upon
which the ideI\tification or the determination of the purity of I:!substance may be based. Frequently, the official standards are
descriptive telms to indicate the approximate solubility of substances, viz.:;(1) "Very soluble," 1 part of the substance is soluble
in less thao/1 part of the solvent; (2) "freely soluble," 1 part of the
substancft'is soluble in from 1 to 10 parts of solvent; (3) "soluble,"
1 part of the substance is soluble in from 10 to 30 parts of solvent;
(4) "spAringly soluble," 1 part of the substance is soluble in from
30 to )00 parts of solvent; (5) "slightly soluble," 1 part of the
subst~nce is soluble in froVl 100 to 1,000 parts of solvent; (6)
"very slightly soluble," 1 part of the substance is soluble in from
1,00Q to 10,000 parts of solvent; and (7) "practically insoluble,"
mor~ than 10,000 parts of solvent are required to dissolve 1 part
of the substance.
There are nine possible classes of solutions, namely: (1) a gas
in a gas; (2) a gas in a liquid; (3) a gas in a solid; (4) a liquid
in a gas; (5) a liquid in a liquid; (6) a liquid in a solid; (7) a
solid in a gas; (8) a solid in a liquid; and (9) a solid in a solid.
Although examples of all of these different types of solutions
are known, the only classes of pharmaceutical importance are
solutions of gases in liquids, olliquids in liquids, and of solids
in liquids.
The official determination of the solubility of gases in water,
such as HCI and NH a, is generally effected by chemical titration
methods, while the solubility of liquids in liquids and of solids
in liquids is determined by physical methods.
203
204
Numerous factors affect the rate and the extent of the solubility of a substance in a given solvent as follows:
1. The solubility of most of the official substances is increased
by a rise in the temperature at which solution is effected. There
are numerous exceptions to this generality, however; e.g., gases,
calcium salts; and ether are less soluble in hot thah in cold water.
Since the solubility of substances varies marketlly with slight
changes in temperature, it is very important that a constant
temperature be accurately maintained throughout a solubility
determination.
2. Substances in a fine state of division dissolve more rapidly
than large crystals or particles because the tot!)'l surface area
exposed to the action lof the solvent
is much greater when. the substance
is powdered.
3. The purity of the ~ubstance and
of the solvent must be assured in all
solubility determinations, since slight
amounts of impurity in either may
cause considerable- variation in the
results. In the official state~ents of
solubility, it is assumed that both the
substance and the solvent employed
conform to the official tests for purity.
4. The position of the solute in the
solvent affects the rate of solution.
If the solute is allowed to lie on, the
FIG. IS.-Apparatus for solubility determinations. A As- bottom of the vessel, it becomes
sembled apparatus. B Stirrer. surrounded by a layer of concentrated
solution which prevents the access of fresh portions of the solvent
to the surface of the solute.
The determination of the solubility of a solid in a liquid necessitates the preparation of a saturated solution. The prod.uction
of a saturated solution of this type may be carried out in the
apparatus illustrated in Fig. 18. This apparatus consists of a
hard-glass test tube A of medium size into wh~ch the solvent and
solute are placed and stirred vigorously by means of a motordriven glass screw stirrer B. The stem of the stirrer passes
through a glass tube, inserted through a well-cleaned stopper by
SOLUBILITY
205
which the solubility tube is closed. The glass tube selected

should be of su~h a size that the stem of the stirrer just passes
through, the area of contact being well lubricated with petrolatum. When splubility determinations are made, the solubility
tube is fastened upright in a thermostat maintained at a constant
temperature of. 25C.
Exercise 68
Object.-To Determine the Solubility of Boric Acid in 'Water

at 25C.
},faterials Required.-1.5 Gm. of boric acid.
A solubility apparatus and thermostat.
Procedure.-1.,'In the solubility tube, place about 1.5 Gm. of finely
powdered boric acid, 25 cc. of distilled water, and after fitting it with the
stirrer, place it inr the thermostat so that it is immersed in the water up to
the level of the stopper, and stir the solution vigorously for about 3 hr.
The boric acid is powdered finely because substances in fine

particles disFlolve more rapidly than do large crystals; subdivision
increases the surface area exposed to the action of the solvent.
The soJubility tube is immersed in the thermostat bath, to
maintaiI). a constant temperature throughout the mixture, since
the solubility of boric acid varies appreciably with slight changes
in temperature. The temperature in the thermostat should notl
vary by more than O.le., above or below 25C., during the
determination. The mixture is stirred continuously to bring
fresbiportions of solvent in contact with the undissolved particles
of boric acid and insure equal concentration throughout the
solution.
2. Stop the stirrer and remove it from the tube, stopper the latter securely,
allow the undissolved crystals to settle, remove about 5 cc. of the clear
solution from the solubility tube by means of a pipette, and weigh accurately.
Evaporate this portion of the solution to dryness on a water bath at 80C.
and finally to constant weight at 900. in an electric or other suitable
mz:en, and weigh the residue.
The weight of the boric acid contained in the residue, subtracted from the weight of the solution analyzed, gives the weight
of the "\Vater in which the boric acid was dissolved.
3. Attach the stirrer, and allow the stirring to continue for about one
hour. Again remove. a 5 cc. portion of the solution, weigh it accurately,
206
and determine the amount of boric acid contained in it in the same manner
as before.
If this result agrees with the former determination, it shows

that saturation of the solution is complete, but ru the amount of
dissolved solid is greater in the second case, the stirring must be
continued .until the concentration of the solution becomes
constant.
4. Express the results as grams of boric acid soluble in 100 Gm. of water.
If a 5.5 Gm. portion of a saturated boric acid solution yielded

0;3000 Gm. of boric acid upon drying to constant weight, the
0.3 Gm. of boric acid was dissolved in 5.5 -.. 0.3 = 5.2 Gm.
water. The amount of boric acid soiuble in 100 Gm. of water at
25C. may therefore be calculated from the simple proportion
0.3:5.2: :X:100
in which X = the number of grams of boric acid ~oluble in 100
Gm. of water at the temperature at which the dEl,termination
was carried out.
Calculate the solubility of boric acid in water at 25C. from the
results of the above determination.
1. What factors influence the rate of solution of boric acid in water?
2. Why is the accurate control of temperature throughout a solpbility
determination essential?
3. Consult the table on page 623 of the U.S.P. and express the restlts of
the above determination in terms of grams of boric acid soluble in 100 cc.
of water at 25C., indicating all calculations, if any are necessary.
CHAPTER XII
SPECIFIC GRAVITY AND DENSITY
The absolute density 'of a solid or liquid is the mass of unit
volume of the substance. Generally, however, in using the term
density, the relative density or the density at a given temperature
relative to the density of distilled water, either at the same temperature, represented by d, or at 4C., represented by dfo is
understood. When in the latter case, the density is corrected
for the buoyancy of the air, the number obtained represents the
true specific gravity of the substance. . In common practice, however, and in all except very exact work, the specific gravity of a
substance is taken as the ratio of the weight of the substance to
the weight of an equal volume of distilled water at the same
temperature.
In the official standards, the specific gravity of a substance has
been established as the ratio of the apparent weight of the substance in air at 25C. to that of an equal volume of distilled water
at the same temperature, or d~~~, except when otherwise stated and
in the case of alcohol, which should be determined at 15.56C.
and compared with water at the same temperature. The latter
is the standard temperature established by the U. S. Revenue
Department for the measurement of the specific gravity of alcohol.
A considerable number of methods hav:e been developed by
which the specific gravities of various substances can be determined. Only those methods of practical importance most
commonly used in pharmaceutical analysis, to establish the
identity and purity of medicinal substances, will be considered
in the following pages. These methods may be classified as
follows:
A. Methods used to determine the specific gravity of liquids.
1. By the use of a pycnometer.
2. By the use of the Westphal balance.
207
208
3. By weighing a solid of known specific gravity and weight

in the liquid.
4. By the use of hydrometers.
B. Methods used to determine the specific gravity of solids,
such as:
1. Solids heavier than and insoluble in water.
2. Solids lighter than and insoluble in water.
3. Solids heavier than and soluble in water.
4. Solids lighter than and soluble in water.
METHODS USED TO DETERMINE THE SPECIFIC GRAVITY OF
LIQUIDS
1. By the Use of a Pycnometer.-The specific gravity of

liquids can be determined very accurately by means of some form
FIG. 19.-Specific gravity bottle.
FIG. 20.-Sprengel-Ostwald
pycnometer (modified form).
of pycnometer or specific gravity bottle, such as that shown

in Fig. 19. Although a number of different forms of pycnometers
have been developed by investigators, those described may be
used .for practically all of the determinations of the specific
gravity of liquids required in pharmaceutical analysis.
The Sprengel-Ostwald Pycnometer.-One of the simplest and
most generally usefuf forms of pycnometers is that known as
the Ostwald modification of the Sprengel tube (Fig. 20). This
form of pycnometer, which can be easily made by the student,
consists of a thin glass tube, 10 to 15 mm. in diameter, to which
narrow tubes, a and b are sealed and bent as shown in the figure.
At about the middle of tube b, the bore should be slightly constricted and a graduation mark scratched or etched on the glass.
Tube a should be drawn out to a point and fire polished so that
SPECIFIC GRA VITY AND DENSITY
209
the bore is constricted. Some of the commercially available

pycnometers of this type (Fig. 21) are provided with ground-on
glass caps which fit over the ends of the tubes A and B. In all
except very accurate work, however, these caps are unnecessary.
The volume of the pycnometer should be about 5 to 15 cc.,
thus permitting of ,an accuracy of about one unit in the fourth
decimal place. When only a very small volume of liquid is
available for a determination, however, a tube of small bore bent
in the shape indicated in Fig. 20, and of a capacity of 0.5 cc.,
may be used in approximate specific gravity determinations.
In carrying out a determination with this type of pycnometer,
it should be cl~ned by washing successively with water, alcohol,
O~~A~
IfPE=B~~
FIG. 21.-Sprengel-Ostwald pycnometer with ground-on glass caps and groundon glass side tube for use in filling.
and ether and then dried by drawing.a current of warm air

through the tube. The pycnometer, cleaned and dried, is first
weighed empty. To do this, it is suspended from the end of a
balance beam by means of a double hook made from either platinum or copper wire, preferably the former. The pycnometer
is then filled with distilled water, which has been previously
boiled to expel dissolved carbon dioxide and allowed to cool,
by applying gentle suction through a rubber tube attached to
the tube b while the tip of the tube a dips into the water. The
introduction of air bubbles into the pycnometer must be carefully
avoided. The filled pycnometer is then suspended in a beaker
of water, along with a thermometeli, and the temperature is
kept constant at the desired lever.
When the pycnometer and its contents have acquired the same
temperature as the bath, usually in about 20 min., the amount of
water must be so adjusted that it exactly fills the pycnometer
from the point of the tube a to the graduation mark on b. If
there is too much water in the pycnometer, a piece of filter
210
paper is carefully touched to the tip of the tube a, thereby

drawing water from the pycnometer until the meniscus corresponds to the mark on b. If the pycnometer is not completely
filled, a rod or tube carrying a drop of water is touched against
the, end of the tube a, and water is drawn into the tube by
capillarity.
The pycnometer is removed from the bath and the outside is
dried with a cloth, using care to prevent the expulsion of water
from the tube by expansion due to the heat of the hands, and
weighed.
After the pycnometer filled with water has been weighed, it
is emptied, dried once more, and filled with the li~uid the specific
gravity of which is to be determined. It is placed
in the constant temperature bath, the volume of
liquid is adjusted to the mark, the pycnometer is
dried with a cloth in the same manner as before and
weighed.
The Geissler pycnometer or specific gravity bottle
(Fig. 22) is a type of pycnometer well suited to
accurate determinations and can- be procured in
sizes varying from 25 to 100 cc. capacity. Pycnometers of this type are provided with al thermometer stopper and a capillary overflow tube fitted
with a ground glass cap to prevent evaporation.
The cap should have a very small perforation in its
top to permit the liquid to expand without forcing
FIG. 22.- the cap loose.
The thermometers provided with
G e i B B 1 e r, these pycnometers are not always accurate and
pycnometer. soul
h d b e teste d
. h one 0 f k nown
y b
companson
WIt
accuracy whenever exact determinations are made.
In ascertaining th,e specific gravity of a liquid with this
pycnometer, the thermometer and cap are removed, the previously cleaned bottle is filled with the liquid which has been
cooled to from 1 to 3C. below the temperature at whicli the
specific gravity is to be determined. The thermometer is then
fitted to the bottle, using due precautions so that no air bubbles
remain in the bottle. The pycnometer with its contents is
warmed to the required temperature, any excess liquid that
exudes from the capillary tube is carefully wiped off, the cap
is fitted to the side tube and the whole is weighed.
211
The official procedure for the determination of the specific

gravity of a fat or oil by the pycnometer method and which may
be applied to most liquid substances is as follows:
"The specific gravity of a fat or oil shall be determined at
250., except when the substance is a solid \1t that temperature.
In this case the specific gravity shall be determined at the
temperature directed in the respective monograph and referred
to water at 250.
"Olean a pycnometer (use a Sprengel pycnometer or a Squibb
pycnometer with a well-fitted capillary stopper) by filling it
with a saturated solution of chromium trioxide or potassium
dichromate in sulfuric acid and allowing it to stand for at least
four hours. Empty the pycnometer and rinse it thoroughly with
distilled water; then fill it with recently boiled distilled water
previously cooled to about 200. and place in a constant temperature bath at 250. At the end of thirty minutes adjust the level
of the water to the proper point on the pycnometer; put the
perforated cap or stopper in place; remove from the bath; wipe
dry with a clean cloth, free from lint; and after allowing to stand
for thirty minutes, weigh. Empty the pycnometer, rinse several
times with alcohol and then with ether, allow it to become
perfectly dry, remove any ether vapor, and weigh. Ascertain
the weight of the contained water at 250. by subtracting the
weight of the pycnometer from its weight when full.
"Fill the clean, dry pycnometer with the oil at a temperature
below.that at which the determination is to be made; place it in
a constant temperature bath at the specified temperature for
thirty minutes; adjust the level of the oil to the proper point
on the pycnometer; put the cap or stopper in place, wipe dry;
allow to stand for thirty minutes; and weigh. Subtract the
weight of the empty pycnometer from its weight when filled
with oil, and divide the difference by the weight of water contained at 250. The quotient is the specific gravity at the temperature of observation, referred to water at 250."
Exercise 69
To Determine the Alcohol Content of an Official Preparation.It is very difficult to effect a quantitative separation of alcohol
from the solvents ordinarily employed in the manufacture of
pharmaceutical preparations, but it is comparatively easy to
212
prepare an aqueous distillate containing all of the alcohol in the

original mixture and to estimate the alcohol in the distillate by
some physical means, such as the determination of the specific
gravity, the index of refraction, or the boiling point of the solution. The specific gravity method is official, and it is probably
the most accurate and satisfactory of the methods mentioned
above. This method consists of separating the alcohol from
substances other than water contained in the preparation by
distillation or other procedures and determining the per cent of
alcohol in the preparation from the specific gravity of the distillate. The following notes which deal with the methods of
treating various types of preparations are taken from the U.S.P.:
"General Process.-Measure accurately not less than 25 cc.
of the liquid in which the alcohol is to be determined, and note
its temperature. Transfer it to a suitable distilling apparatus
and, if its alcohol content is thought to be not more than 30 per
cent, dilute it with an equal volume of distilled water, using the
water to rinse the vessel that was used for measuring. Distil,
and collect a volume of distillate of about 2 cc. less than the
volume of the original test liquid, cool or bring ~o the temperature
at which the original test liquid was measured, add sufficient
distilled water to measure exactly the original volumf of the test
liquid and determine the specific gravity. From this result
ascertain the percentage, by volume, of alcohol contained therein.
The proportion of alcohol by volume in the distillate equals
that in the liquid examined.
"If the liquid under examination contains more than 30 per
cent of alcohol, proceed as directed above, except: dilute the
sample with about twice its volume of distilled water and collect
a volume of distillate of about 2 cc. less than twice the volume
of the original test liquid, cool or bring to the temperature at
which the original liquid was measured, add sufficient distilled
water to measure exactly twice the original volume of the test
liquid and determine the specific gravity. The proportion
of alcohol by volume in this distillate, as ascertained from its
specific gravity, equals one-half that in the liquid examined.
"The distillate must be clear or only slightly cloudy, must not
contain any non-volatile material and not more than traces of
volatile substances other than alcohol and water.
213
"This general method without modification is suitable for

examining most fluid, extraQts and tinctures provided the
capacity of the distilling flask is sufficient (commonly two
to four times the volume of the liquid to be distilled) and the rate
of mstillation is such that clear distillates are produced. Some
alcoholic preparations will, however, require special treatment
or the observance of special precautions to yield suitable distillates. All distillates must be clear or nearly so.
"Frothing.-Liquids which froth to a troublesome extent during
distillation may be distmed by strongly acidulating them with
phosphoric or sulfuric acid, or by the addition of a slight excess
of calcium chloride solution or by the addition of a little paraffin
or yellow wax to the distilling flask.
"Bumping.-Liquids that tend to bump badly when heated
(particularly resinous solutions) may be distilled by making them
alkaline with magnesia magma or by placing pieces of pumice
or angular pieces of metal in the distilling flask with the liquid,
or by similar means of distributing the heat.
"Glycerin.-Liquids that contain glycerin must be diluted
with sufficient distilled water so that the residue, after distillation,
will contain at least 50 per cent of water.
"Iodine.-All Solutions of Iodine must be decolorized, i.e.,
deprived of free iodine, before being distilled, by treatment with
powdered zinc or by the addition of a solution of sodium thiosulfate. In the latter case a large excess of the thiosulfate must
be avoided and must be followed by a few drops of sodium
hydroxide solution to fix volatile sulfur compounds.
"Volatile Substances.-Spirits, elixirs, tinctures, etc., that
contain appreciable proportions of volatile materials other than
alcohol and water, such as volatile oils, chloroform, ether,
camphor, etc., are treated as follows: Mix the accurately measurcd liquid with about an equal volume of saturated solution of
sodium chloride in a separator, add about the same volume of
"purified petroleum benzin and shake the mixture to extract the
interfering volatile ingredients. Draw off the separated lower
layer and extract the benzin solution with two successive portions
of a saturated solution of sodium chloride, using about one-half
as much each time as was used in the first extraction mixture.
Combine the aqueous saline solutions and distil the mixture in
214
the usual way, collecting a volume of distillate having a: simple

ratio to the volume of the origin~lliquid.
"A troublesome emulsion may develop in the liquid mixture
upon shaking it with the benzin. In such case dilute a fresh
portion of the original liquid with water and distil it as directed
in the general process. Then treat this distillate as directed
above, using purified petroleum benzin and sodium chloride
solution, and distil the aqueous saline solution so produced, to
obtain a distillate which is free from volatile substances other
than alcohol and water.
"In preparing collodion for distillation, water is used in place of
the saturated solution of sodium chloride directed above.
"If volatile oils are present in small proportions only, and a
cloudy distillate is obtained, the benzin treatment not having
been employed, the distillate may be clarified and rendered
suitable for the specific gravity determination by shaking it
with about one-fifth its volume of purified petroleum benzin, or
by filtering it through a thin layer of purified talc.
"Other Preparations Requiring Special Treatment.-Preparations containing free ammonia, as Aromatic Spirit of Ammonia,
must be rendered slightly acid with sulfuric acid before being
distilled.
I
"Spirit of Ethyl Nitrite, Spirit of Glyceryl Trinitrate and
Aromatic Sulfuric Acid must be treated with a small excess of
sodium hydroxide before being distilled. The use of this expedient in examining Spirit of Ethyl Nitrite necessitates that a suitable correction be subtracted from the apparent result of the
alcohol determination to compensate for the alcohol produced
during the decomposition of the ethyl nitrite.
"Chloroform Liniment, Camphor and Soap Liniment, and
Liniment of Soft Soap are treated with an excess of sulfuric acid
to effect decomposition of the soap before they are extracted with
purified petroleum benzin as directed in the general process."
Object.-To Determine the Alcohol Content of Fluidextract of
Belladonna Leaf.
Materials Required.-lO cc. of fluidextract of belladonna leaf.
A distilling apparatus.
A flask graduated to hold 50 cc.
A pycnometer.
215
Procedure.-l. Place 10 cc. of the preparation, measured at 25C., in a

distilling flask of about 200 cc. capacity, provided with a high side tube connected with a suitable condenser; add 50 cc. of distilled water, distil into a
50 cc. graduated flask, at such a rate that about 48 cc. of distillate will be
received in 72' hr.; if volatile products other than alcohol are known to be
absent, bring to the original temperature and dilute to 50 cc. with water of
the same temperature.
In very accurate work, it is best that the volume of the sample

and of the distillate be measured at the same temperature, i.e.,
15.56C. When the temperatures differ, the volume of the
sample used and of the distillate collected are not the same,
since the coefficient of expansion of alcohol is much greater than
that of water. In the volume percentages for alcohol, it is
assumed that the volumes are measured at 15.56C.
2. Determine the specific gravity of the distillate at 25C. by means of a
pycnometer, and find the per cent of absolute alcohol by volume by reference to the alcoholometric table in the U.S.P., page 604, and multiply by
the dilution factor to find the per cent of alcohol in the original pteparation.
The specific gravity of the distillate may be determined by

means of either the Sprengel-Ostwald or the Geissler pycnometer
as previously described. The latter instrument is the more
satisfactory, since the temperature of the alcoholic solution can
be determined while the specific gravity determination is being
made.
The method of recording the data and calculating the results is
illustrated in the example:
Volume of sample at 25C. = 10 cc.
Volume of distillate at 25C. = 50 cc.
Dilution factor = 5%0 = 5
The specific gravity of the distillate at 25/25 found = 0.9844.
By referring to the alcoholometric table, page 604 of the
U.S.P., the percentage by volume of alcohol in the distillate is
found to be 11 + 0.5 = 11.5. " This percentage, is based on the
apparent specific gravity of alcohol at 15.56/15.56. The
per cent of absolute alcohol by volume contained in the sample
tested is found by multiplying the per cent found in the distillate
by the dilution factor, or 11.5 X 5 = 57.5 per cent. The
corresponding per cent of absolute alcohol by weight may be
216
found by consulting the table on page 604 of the U.S.P.; e.g.,

57 and 58 per cent alcohol by volume are found to correspond to
49.19 and 50.17 per cent alcohol by weight, respectively. Therefore, 57.5 per cent alcohol by volume corresponds to 49.68 per
cent by weight.
Toe amount of alcohol present in various preparations such as
fiuidextracts and tinctures is subject to considerable variation
due to the varying proportions of moisture and extractive contained in the drug used and to the methods of extraction and
FIG. 23.-Plummet.
FIG. 24.-Westphal specific gravity balance.
FIG. 25.-Riders.
concentration employed in their preparation. The official

standards, therefore, generally give a range of alcohol content for
preparations of this type; e.g., fluidextract of belladonna leaf is
required to contain 57 to 63 per cent of alcohol by volume.
2. By the Use of the Westphal Balance.-The specific gravity
of liquids may be determined quickly and with a fair degree of
accuracy by means of the apparatus known as the Westphal
balance. The mechanism of this device is based upon the generally known law of physics that a body immersed in a liquid is
buoyed up by a force equal to the weight of the liquid displfwed.
This apparatus (Figs. 23, 24, an~ 25) consists of a beam balanced
on a knife edge A with a sinker or plummet B, suspended from
one end and counterpoised by a fixed brass weight C at the other.
The distance between the knife edge A and the plummet support
D is graduated into ten divisions of equal length. The plummet,
217
which is made of glass and is provided with a thermometer, is

made of such size that it will displace exactly 5 Gm. of water at a
given temperature, usually 15.56C. The plummet and the
platinum wire by which it is suspended are .usually made of a
definite weight so that they are interchangeable on different
balances.
Exercise 70
Object:-To Determine the Specific Gravity of a Volatile Oil.

Materials Required.-A Westphal balance.
A 50 cc. cylindrical graduate.
50 cc. of volatile oil.
Procedure.-l. Balance the apparatus in air.
To balance the apparatus, place it upon a firm, level t~ble

and be sure that the leveling screw in the base is directly beneath
the arm which supports the plummet, compare the height of the
beam with that of the cylinder which is to contain the oil, lengthen
or shorten the standard if necessary, and adjust the level of the
beam by means of the leveling screw E until the points F and G
exactly coincide.
2. Suspend the plummet in wate-r, contained in the glass cylinder, place
the largest rider weight on the hook to which the plummet is attached, and
balance the instrument again by turning the leveling screw.
When determinations are made at the exact temperature for

which the instrument is designed, no adjustment should be necessary at this point, but in taking the specific gravity at 25C.,
a slight adjustment is required to correct for the expansion of the
liquid at the higher temperature.
3. Dry and clean the plummet and cylinder, fill the cylinder with the oil
to the same level as that reached by the water in balancing the apparatus,
lift the plummet and place the cylinder beneath the end of the beam,
replace the plummet so that it hangs free in the oil without touching the
cylinder wall, and place the riders ~n the beam until balance is restored.
The temperature of the oil (25C.) at which the final adjustment of the balance is made should be the same as that of the
water used to standardize the apparatus. The specific gravity
is read directly from the position of the riders on the beam;
e.g., if the largest rider is at 9, the second at 7, the third at 5,
218
and the fourth at 2, the specific gravity of the oil is 0.9752.

Readings should always be made to four decimal places; i.e.,
all four sizes of riders should be used.
3. By Weighing a Solid in Air, in Water, and in the Liquid.Plummets or sinkers of various forms made of sealed glass tubes
partly filled with mercury or of some metal such as platinum
have been devised for specific gravity determinations of this
type, but a brass weight will suffice for the determination of the
specific gravity of liquids which do not attack the brass. For
example, if a piece of brass weighs 10 Gm. in air, 8.824 Gm.
in water, and 9 Gm. in a liquid of unknown specific gravity, the
specific gravity of the last can be calculated from a simple
proportion as follows:
1.176:1::1':X
x =
0.85
where 1.176 is the loss of weight in water (10 - 8.824 = 1.176)

or the weight of the water displaced; 1 is the loss of weight in the
liquid;,l', the specific gravity of water; and X, the specIfic gravity
of the unknown liquid.
The principles involved and the method of making the determination are similar to that given in detail under the determination of the specific gravity of solids heavier than anp. insoluble
in water (page 219).
4. By the Use of Hydrometers.-Hydrometers are instruments
used to determine quickly the approximate specific Igravity or
density of liquids by flotation. They usually consist of a sealed
glass tube weighted with mercury or shot and frequently are
provided with a thermometer so that the temperature at which
the specific gravity is determined may be read at the same time.
They are variously called alcoholometers, lactometers, urinometers,
etc., according to the use for which they are designed. They are
either constructed to read specific gravity directly, as, for
example, the hydrometers for general use and lactometers, or they
are graduated according to some arbitrary scale of degrees, as
in the case of the Baume and Twadell hydrometers. Specific
gravity hydrometers are usually made in sets of four or six, each
instrument being calibrated to cover a different range in specific
gravities so that the complete set will suffice to determine the
specific gravity of almost any liquid; e.g., the different instru-
219
ments in the set of four are calibrated to read from 0.600 to 1.000,
from 1.000 to 1.400, from 1.400 to 1.800, and from 1.800 to 2.200.
When making a specific gravity determination with a hydrometer, the thoroughly cleaned instrument should be allowed to
sink in the liquid gradually until it is at rest, then depressed
about 1 in. to 'moisten a portion of the stem, allowed to come to
equilibrium, and the reading taken.
Exercise 71
Object.-To Prepare 200 cc. of Sulfuric Acid Containing

10 Gm. of H 2 S0 4 per 100 cc.
Materials Required.-20 cc. of concentrated H 2S0 4
A hydrometer calibrated from specific gravity 1:000 to 1.400 at 250.
Procedure.-Dilute 20 cc. of pure concentrated sulfuric acid to 100 cc.
with distilled water, allow the resulting solution to acquire a temperature
of 250., and take its specific gravity as accurately as possible with the
hydrometer.
If the specific gravity of the solution is found to be 1.2603,

it contains 35 per cent of H 2S0 4 as found by referring to the
table, page 615 of the U.S.P. The weight of the prepared acid
solution to be diluted will then be 10 X
~5 X 100
= 57.1430 Gm.
This weight of solution containing 35 per cent of H 2S0 4 when

diluted to 200 cc. will form a solution containing 10 Gm. of
H 2S0 4 per 100 cc.
Solutions prepared as directed above are subject to slight
inaccuracies of concentration, since specific gravity determinations by means of hydrometers are not exact, but if the specific
gravity is determined accurately by means of a pycnometer,
the concentration of a solution prepared in this manner will be
sufficiently accurate for all ordinary qualitative purposes.
METHODS USED TO DETERMINE THE SPECIFIC GRAVITY OF
SOLIDS
1. Solids Heavier than and Insoluble in Water.---:-A. Weigh

the substance in air and then in water.
According to the law of Archimedes, a body suspended in a
fluid is j:moyed up by a force equal to the weight of the fluid
displaced. The weight of a solid in air, therefore, less its weight
220
in water gives the weight of an equal volume of water, since

the volume of water displaced is equal to the volume' of the
solid. If the weight in air is divided by the loss of weight in
water, the result is the specific gravity of the substance at the
temperature at which the determination is made. For example:
Suppose that a piece of brass weighs 10.0040 Gm. in air at 25C.
and 8.8010 Gm. immersed in water at 25C. The weight in air
divided by the loss of weight, 1.2030 Gm., gives the specific
gravity of the brass at 25/25 or 10.0040/1.2030 = 8.32.
B. If the substance is in small pieces or is a powder, the specific
gravity may be determined as illustrated in the following exercise:
Exercise 72
Object.-To Determine the Specific Gravity of Powdered

Barium Sulfate.
Materials Required.-7 to 10 Gm. of powdered barium sulfate.
A small volumetric flask (25 or 50 cc.).
Procedure.-l. Weigh about 10 Gm'. of the powdered barium sulfate
accurately.
2. Accurately weigh the small volumetric flask filled exactly to the
graduation mark with recently boiled distilled water at"25C.
3. Introduce the powdered barium sulfate into the flask quantitatively,
remove any air occluded in the powder by agitating the mixture\ draw off the
water with a pipette until the level of the liquid is again exactly at the
graduation mark when the liquid is at 250., and weigh the flask and its
contents accurately.
It is evident that a volume of water equal to that of the barium

sulfate has been displaced and removed from the flask. The
specific gravity of the barium sulfate may then be calculated, e.g.:
Weight of powdered barium sulfate = 8.0 Gm.

Weight of flask filled with water = 80 Gm.
Weight of flask, barium sulfate, and water = 85 Gm.
80 + 8 - 85 = 3 Gm., the weight of the volume of water
displaced by the barium sulfate; and % = 2.6666, the specific
gravity of the barium sulfate.
2. Solids Lighter than and Insoluble in Water.-A. By Weighing in Water.-If a substance is lighter than water, it will float,
and its volume cannot be obtained by weighing it in water unless
it is completely submerged by means of a weight or sinke~ heavier
221
than water. The method of determining and calculating the

specific gravity of a substance of this type is illustrated in the
following exercise:
Exercise 73
Object.-To Determine the Specific Gravity of Camphor.

Materials Required.-A piece of camphor (2 to 4 Gm.).
A suitable sinker such as a 2 Gm. weight.
Procedure.-l. Weigh the piece of camphor in air.
2. Weigh the sinker immersed in water.
3. Attach the sinker to the piece of camphor and find their combined
weight when immersed in water.
Since the camphor is lighter than water, the weight of the sinker
and camphor is less than the weight of the sinker alone. The
difference between the weight of the sinker and camphor in water
and the weight of the sinker alone in water plus the weight of the
camphor in air is a measure of the water displaced by the camphor
and also a measure of its buoyant power, e.g.:
Weight of the camphor in air = 4 Gm.
Weight of the sinker in water = 1.6 Gm.
Weight of the camphor and sinker in water
1.5 Gm.
The difference between the weight of the sinker in water and

the weight of the sinker and camphor in water = 1.6 - 1.5
= 0.1 Gm.
The weight of the camphor in air plus this difference of 0.1 Gm.
is the weight of a volume of water equal to the weight of the
piece of camphor or 4
0.1 = 4.1 Gm. water displaced.
Since the specific gravity of a substance is found by dividing
its weight by the weight of an equal volume of water, the specific
gravity of the camphor is 4/4.1 = 0.9756.
B. By the Flotation Method.~This method consists of bringing
the substance into equilibrium with a liquid of known specific
gravity. It is used in the official determination of the specific
gravity of such substances as white and yellow wax and spermaceti and by food and drug analysts to determine the specific
gravities of certain fats and oils.
222

Exercis.e 74
Object.-To Determin.e the Specific Gravity of Yellow Wax.

Materials Required.-About 0.25 Gm. of yellow wax.
25 ce. of alcohol.
A pycnometer.
Procedure.-l, "Melt the Wax at a low temperature and allow it to fall
into separated drops from just above the surface into alcohol that has been
warmed to from 45 to 50C. Allow the globules to remain in the alcohol
until it has cooled spontaneously to room temperature (20 to 25C.), then
remove the Wax and keep it at room temperature for twenty-four hours."
The melted wax separates in globules free from occluded air

which solidify when the mixture is cooled. The wax which has a
high coefficient of expansion is kept at room temperature for
24 hr. to permit the globules to acquire their normal volume.
2. "Prepare a mixture of four volumes of alcohol and enough distilled
water to make ten volumes, and allow it to stand until free from air bubbles.
Moisten the globules of Wax with distilled water by means of a brush, and
place them by means of forceps in the alcohol solution just prepared and
contained in a beaker. Then add alcohol or air-free distilled water, as
required, to the mixture kept at 25C., until the globules of the Wax float
and rest indifferently in the liquid. Finally determrne the specific gravity
of the alcohol-water mixture. The figure thus obtained represents the
specific gravity of the Wax examined."
The wax globules are moistened with water so that the water:alcohol mixture will come in contact with all portions of the
globules without occluding air at the intersurface. When the
globules are in equilibrium in the alcoholic solution, their weight
must be equal to the weight of liquid which they displace or they
would rise to the top of the liquid and float or sink to the bottom
of the beaker. Consequently, when such equilibrium is attained,
the specific gravity of the wax and that of the liquid are equal,
and a determin.ation of the specific gravity of the liquid is a
measure of the specific gravity of the wax.
3. Solids Heavier than and Soluble in Water.-The specific
gravity of solids heavier than and soluble in water may be determined by weighing the solid in air and then in some liquid of
known specific gravity in which the solid substance is insoluble
such as oil of -turpentine, chloroform, benzene, etc. Thus, the
weight of a piece of alum in air divided by its loss in weight when
223
weighed immersed in oil of tu.rpentine gives the density of the

alum relative to the oil of turpentine, and if the specific gravity
of the oil of turpentine is known, that of the alum can be calculated, e.g.:
Weight of the alum in air = 10 Gm.
Weight Of the alum in oil of turpentine = 6.5 Gm.
Specific gravity of the oil of turpentine = 0.860
10 - 6.5 = 3.5 Gm., loss in weight of the alum in the oil of
turpentine equal to the weight of an equal volume of the latter.
3.5:X::0.86:1
= 4.070 Gm.,
.
where X is the weight of a volume of water equal to the volume
of the oil of turpentine displaced, and 10/4.070 = 2.4570, the
specific gravity of the alum.
4. Solids Lighter than and Soluble in Water.-The specific
gravity of substances lighter than and soluble in water may be
de~ermined by weighing the substance with a sinker in some
liquid of known specific gravity in which the substance is not
soluble. For example, the specific gravity of lithium may be
determined from the data below as follows:
Weight of the lithium in air = 2.0 Gm.
Weight of the sinker in air = 5.0 Gm.
Specific gravity of the sinker (brass) = 8.4
Specific gravity of liquid (kerosene) = 0.830
Weight of lithium and sinker in kerosene = 3.8 Gm.
The density X of the sinker referred to kerosene as a standard is
0.830:1 ::8.4:X
10.1205
The weight Y of the sinker in kerosene may be calculated,
5.05~
Y = 10.1205
Y = 4.5095,
and the density of lithium compared to that of kerosene is

2
+ 4.5060
- 3.8 = 0.7391.
224
The specific gravity Z of the lithium is, therefore,

0.830: 1:: Z: 0.7391
Z = 0.6135, the specific gravity of the lithium.
1. Define the terms specific gravity, apparent specific gravity, true

specific gravity.
2. By what different methods might the specific gravity of alcohol be
determined?
3. A 10 Gm. weight of brass, specific gravity 8.4, submerged in glycerin
weighed 8.5135 Gm. What is the specific gravity of the glycerin?
4. Given the specific gravity'of sulfuric acid at 15C. as 1.8422 compared
to water at 15C., what is its specific gravity referred to water at 21C. 7
(See U.S.P., page 617.)
5. How much nitric acid, specific gravity 1.45, and how much water must
be mixed to form 1 l. of nitric acid, specific gravity 1.20?
6. Calculate the specific gravity of a piece of sandalwood from the follow"
ing data:
Weight in air = 10.100 Gm.
Weight of sinker in water = 12.200 Gm.
Weight of sinker and sandalwood in water
1)..900 Gm.
7. How many grams of water will a pycnometer calibrater! to hold 100

Gm. of water at 25C./250. and 760 mm. pressure contain at 1190. and 740
mm. pressure? (See U.S.P., page 622.)
8. Give the specific gravity of alcohol compared to water at 15.560. and
760 mm. pressure in a solution containing 16.5 per cent of alcohol by volume.
(See U.S.P., page 604.)
9. Briefly state how the specific gravity of each of the following might be
determined with comparative accuracy; olive oil, benzene, powdered borax,
granular zinc, and cacao butter.
The approximate specific gravities for many official substances,

such as acetone, acids, and numerous preparations, are given in
the U.S. Pharmacopoeia and National Formulary simply as
information for toe conversion of volume to weight. When the
minimum or maximum specific gravity or a range of specific
gravity is given, it is a definite constant of value in
determining the identity, purity, and concentration of the
substance. Those substances for which definite specific gravity
limits are given in the official standards are shown in the following
table.
225

TABLE XXX.-OFFICIAL SUBSTANCES WITH SPECIFIC GRAVITIES
Substance
U.S.P.
Acid, aeatic, glacial. ......
Alcohol (at 15.56C.). , .
Al c oh 01, d eh yd r a ted
(15.56C.) ..............
Alcohol, diluted (15.56C.).
Amyl nitrite............
Balsam, Peruvian .........
Benzin, petroleum, purified
Brandy ..................
Bromine, R.1 .............
Camphor, spirit of ........
Carbon tetrachloride ......
Chloroform ..............
Collodion ................
Copaiba .................
Creosote .................
Creosote carbonate ........
Cresol. . . . . . . . . . . . . . . . . ..
Ether ...................
Ether, absolute, R ........
Ethyl acetate, R ..........
Ethyl nitrite. spirit of. ....
Ethyl oxide ..............
Eucalyptol. ..............
Eugenol. . . . . . . . . . . . . . . ..
Ferric chloride, solution of
Glycerin .................
Giyceryl trinitrate, spirit of
Honey' ..................
Malt, extract of. .........
Methyl salicylate (natural)
Methyl salicylate (synthetic) ....................
Oil (fixed) of
almond, expressed .......
castor .................
chaulmoogra ...........
cod liver ...............
corn ...................
cottonseed .............
linseed .................
olive ....... \ ..........
theobroma' ............
Oil (volatile) of
anise ..................
bitter almond. . . . . . . . ..
chenopodium ...........
cinnamon ..............
clove ..................
coriander ..............
Specific
gravity,
25C.
Substance
U.S.P.
dwarf pine needles ......
eucalyptus .............
!en~el. ................
(n.m.t.) 0.798
Jumper ................
0.935 to 0.937
lavender .............
0 865 to 0.875
lemon .................
1.150 to 1.170
mustard ...............
0.634 to 0.660
myristica ..............
0.933 to 0.921
orange .. : ..............
(n.l.t.) 3.099
peppermmt ............
0.824 to 0.826
rose' .................
1.588 to 1.590
rosemary ..............
1.474 to 1.478
santa!....... ........
0.765 to 0.775
sassafr'!s. . . . .. ........
0.930 to O. 995
spearmmt .. '" ........
(n.l.t.) 1.076
tar, rectified ............
(n.l.t.) 1.145
turpentine .............
1.030 to 1 .038
turpentine, rectified.. ..
0.713toO.716 Oleoresin of aspidium ......
(n.m.t.) 0.710
Ox bile ..................
0.893 to 0.898 Paraffin, chlorinated .......
(n.m.t.) 0.823
Petroleum' ...............
0.713 to 0.716 Pet~olatum,liquid ........
0.921 to 0.923 Rosm...................
1.066 to 1.070 Spermaceti ...............
1. 29 to 1. 32
Sodium hypochlorite, di(n.l.t.) 1.249
luted solution of ........
0.814 to 0.820 Tar, juniper ..............
(n.Lt.) 1.099
Terebene ................
1.350 to 1.430 Wax, white and yellow ....
1.176 to 1.182 Whisky ..................
N.F.
1. 180 to 1.185 Anethol. ... : ..............
Ether, spmt of ...........
0.910 to 0.915 Ethyl acetate .............
0.945 to 0.965 Caramel.................
0.940 to 0~960 Hamamelis water...
0.918 to 0.927 Oil (fixed) of
0.914 to 0.921
croton ...............
0.915toO.921
sesame ................
0.925 to 0.935 Oil (volatile) of
0.910 to 0.915
birch tar. rectified .... '"
0.858 to 0.864
bitter orange ...........
caraway ...............
0.978 to 0.988
cardamom . . . . . . . . . . .
1. 038 to 1. 060
myrcia ................
(n.l.t.) 0.950
orange flowers ..........
1.045 to 1.063
pimenta........ ....
1.038 to 1.060
thyme .................
0.863 to 0.875 Tetrachlorethylene . . . . . .
1.047 to 1.050
(n.m.t.) 0.816
(n. m. t.) = not more than. (n. I. t.) = not less than. R. = reagent.
1 Saturated with water.
, Diluted with 2 times its volume of distilled water.
, At 100C. compared to water at 25C.
'At 30C.' compared with water at 15C .
At 60C.
Specific
gravity.
25C.
0.853 to 0.869
0.905 to 0.925
0.953 to 0.973
0.854 to 0.879
0.875 to 0.888
0.849 to 0.855
1.013 to 1.020
0.859 to 0.924
0.842 to 0.846
0.896 to 0.908
0.848 to 0.863
0.894 to 0.912
0.965 to 0.980
1. 065 to 1.077
0.917 to 0.934
0.960 to 0.990
0.854 to 0.868
0.853 to 0.862
(n.l.t.) 1.0
1.015 to 1.025
1.00 to 1.07
0.820 to 0.865
0.828 to 0.905
1.07 to 1 09
0.938 to 0: 944
(n.Lt.) 1.205
0.950 to 1.055
0.860 to 0 865
0.950 to 0:960
0.935 to 0.923
0.983 to 0.987
0.784 to 0.794
0.892 to 0.898
(n.l.t.) 1.35
0.979 to 0.982
0.935 to 0.950
0.916toO.921
0.886
0.845
0.900
0.917
0.962
0.863
1.018
0.894
1.600
to
to
to
to
to
to
to
to
to
0.950
0.851
0.910
0.947
0.990
0.880
1.048
0.930
1.610
CHAPTER XIII
MELTING, CONGEALING, AND BOILING POINTS

Melting Point.-When an amorphous substance is heated, it
gradually softens and eventually acquires the properties characteristic of a liquid, but no definite point of transition from the
solid to the liquid state occurs during the process of heating.
When a crystalline solid is heated, there is a sharp change from
the solid to the liquid state at a definite temperature, this temperature being known as the melting point of the substance.
The melting point ora compound may be defined as the temperature at which the liquid and solid phases of the compound are in
equilibrium. Each pure fusible compound
has a fixed and definite melting point, and
this constant is often ...used to determine
the purity of a solid, since the introduction
of even a small amount of impurity lowers
the melting point to a considerable extent.
If appreciable quantities of impurities are
present, the melting point is not sharp but
ranges ove~ a number of degrees and is
usually low. In the official standards, the
melting point of a substance is defined as
the interval of temperature within which
2
the substance is observed to melt when
treated according to a definite method.
FIG. 26.-ApparatuB ferr F
taking melting points.
or the determination of the melting point,
the official substances are divided into two
classes, namely: (1) substances which can be readily reduced to
a powder; (2) substances such as fats, fatty acids, waxes, and
paraffin which cannot be reduced to a powder readily.
The apparatus (Fig. 26) required to determine the melting
point of either class of substances consists of the following parts:
(1) a thin-walled test tube about 3 or 4 cm. in diameter and about
226
227
10 cm. long; (2) a stirring rod bent into a circle at one end to fit
the above tube and at an angle at the other end to permit easy
manipulation; (3) a standard th~rmometer covering the desired
range of temperature; (4) an auxiliary thermometer to take the
emergent stem correction, preferably graduated from 20 to
100C.; (5) a capillary glass tube about 6 cm. long and 1.0 mm.
in diameter sealed at one end.
For temperatures up to 200C., a purified, concentrated sulfuric
acid is a suitable bath. For higher temperatures, up to about
350C., a pure grade of cottonseed oil (almost colorless) will
serve for a limited number of determinations. Other, though
less desirable, substitutes for sulfuric acid for use at high temperatures are: (1) a pure grade of paraffin which has been freshly
distilled; (2) clean, white, hydrogenated cottonseed oil. A very
satisfactory bath is prepared by cautiously boiling toge~her, for
from five to ten minutes under a hood, a mixture of 70 parts of
sulfuric acid and 30 parts of potassium sulfate, stirring constantly
until the potassium sulfate is completely dissolved.
The following exercise illustrates the methods used to determine the melting points of official substances:
Exercise 76
Object.-To Determine t_he Melting Point of Salicylic Acid.

Materials Required.-A melting point apparatus as described above.
20 to 30 cc. of concentrated sulfuric acid.
About 0.05 Gm. of salicylic acid.
Procedure.-l, Reduce the salicylic acid to a fine powder and dry it at
100C. for 2 hr.
The material is dried because substances which contain water

of crystallization or moisture must be rendered ,anhydrous to
prevent liquefaction when heated through the solvent action of
water.
2. Place sufficient sulfuric acid in the test tube to fill it to a depth of
about 5 cm. so that the upper portion of the thermometer bulb may be
immersed about 2 cm. below the upper surface of the bath and leave about
2 cm. of the liquid below the lower end of the bulb.
This arrangement prevents unequal heating of the different

parts of the thermometer bulb.
228
3. Introduce enough of the finely powdered dry salicylic acid into thc
capillary tube to form a column about 0.3 cm. in length.
The salicylic acid may be packed into the sealed end of the
tube by tapping the open end of it into a little mound of the powder, then inverting the tube and tapping it gently on a solid
surface, or by drawing the broad side of a triangular file across
the surface of the tube just below the powder.
4. Attach the charged capillary tube to the thermometer by wetting
both with the acid of the bath, and the tube will then be held in position by
capillary attraction, or attach the capillary tube by means of a rubber band,
clipped from a piece of rubber tubing, in such a position that the column of
salicylic acid is centrally located by the side of the thermometer bulb.
Attach the auxiliary thermometer so that the center of its bulb is as close as
possible to the stem of the main thermometer at a point midway between
the surface of the bath and the 160 graduation mark. Heat the acid bath
by means of a free Bunsen flame until a temperature of about 140C. is
reached, then carefully regulate the rise in temperature to about 3 per
minute until the substance begins to melt, and then regulate the rise in
temperature to about 0.5 per minute until the salicylic acid is completely
melted, while stirring the bath continually. Record the melting interval
temperature and the temperature registered on the auxiliary thermometer
at the end of the melting.
The temperature at which the column of salicylic acid is

observed to collapse definitely in the capillary tub'e is considered
as the beginning of the melting, the temperature at which the
salicylic acid becomes liquid throughout is considered as the
end of the melting, and the interval between the temperatures
at which the melting begins and ends is taken as the temperature
range of the melting point of salicylic acid.
The rate of rise in the temperature of the bath can be regulated
easily if .the burner is held in the hand so that more or less heat
can be applied as required. Since heat from the burner may
affect the temperature registered on the auxiliary thermometer,
it is advisable to construct a movable platform of stiff paper
around the main thermometer and adjust it to a position just
below the bulb of the auxiliary thermometer.
The temperature observed on the auxiliary thermometer is
used to correct for the contraction of the thread of mercury above
the bath by the following formula:
Correction = 0.00015 X N(T - t) where N represents the
number of degrees from the surface of the bath to the melting
229
point, T the temperature at the end of melting, and t the temperature registered on the auxiliary thermometer. The correction is
added to the observed reading of the main thermometer; i.e.,
if in the determination of the melting point of salicylic acid, the
main thermometer is immersed in the bath to the 30 graduation
mark, the acid ~s completely melted at 156.5C. and the temperature registered on the auxiliary thermometer is 40C., the correction for the melting point temperature would be
0.00015 X 126.5(156.5 - 40) = 2.59C.
and the corrected melting point becomes 156.5 + 2.59 = 159.1C.
The melting point of a substance which cannot be readily
reduced to a powder is determined in the same manner with the
following exceptions: The substance is melted at as Iowa temperature as possible and drawn into a capillary tube, open at
both ends, to a depth of about 1 cm. The charged tube is allowed
to cool at 10C. or at a lower temperature for 24 hr. or in contact
with ice for 2 hr. The melting point of the substance is then
determined by attaching the charged tube to a standard thermometer as described in the above exercise or by means of a
rubber band and heating in a suitable bath so that the rate of rise
of temperature does not exceed 0.5 0 per minute for the last 5
before the expected melting point is reached. The temperature
at which the substance is observed to rise in the capillary tube
is taken as the melting point. Melting point determinations
by the above method are generally performed on substances
which are mixtures and which do not give the sharp melting
point characteristic of pure organic compounds.
1. Look up the melting points given for each of the following substances
in the U.S.P. and explain briefly why an interval of melting point temperatures is permitted in some cases while fixed melting points are given in
others: (a) paraffin, (b) petrolatum, (0) acetylsalicylic acid, (d) menthol,
(e) cocaine.
2. Why should thermometers for use in melting point determinations be
made of annealed glass? (See U.S.P., page 470.)
3. How might the emergent stem correction for the main thermometer be
determined by interpolation from a correction curve when a number of
melting points are to be determined? (See U.S.P., page 457.)
230

TABLE XXXI.-MELTING POINTS OF OFFICIAL SUBSTANCES
Substance
Melting
point.
Melting
point,
Substance
C.
U.S.P.
Acetanilid ................
Acetphenetidin ....... , ..
Acid, acetylsalicylic.... ..
Acid, benzoic .............
Acid. salicylio ....... , ....
Aminopyrine ............
Antipyrine ...............
Atropine .................
Atropine Bulfate. . . . . . . . . .
Barbital. ................
Benzidine, R .... , ........
BetanaphthoL ... " .......
Caffeine .................
Camphor ................
Carbromal ...............
ChlorbutanoL. . . . . . . . . . . .
Cocaine. . . . . . . . . . .. .....
Cocaine hydrochloride.....
Codeine (anhydrous) ......
Colchicine ...............
Dichloramine. . . . . . . . . . . . .
Dinitrophenylhydrazine R.
Diphenylamine, R. . . . . . ..
Ephedrine. . . .. . . . . . . . . ..
Ephedrine hydrochloride ..
Ephedrine sulfate .........
Ethyl ..minobcnzoatc. . . . ..
Ethyl morphine hydrochloride ............... .
Guaiacol ............... ..
Histamine phosphate ..... .
Homatropine hydrobromide... . . . . . . . . . . . . . . .
Iodoform.. . . . .. . . . . . . . . .
rsatin, R .................
Lard ................... ,
Menthol. . . . . . . . . . . . . . . ..
Methenamine. . . . . . . . . . . .
Neocinchophen. " . . . . . . . .
Oil. theobroma. . . . .. ....
Paraffin.
Petrolatum ...............
Phenacaine ...............
Phenacaine hydrochloride.
C.
U.S.P. (Oont.)
113 to 115
Phenobarbital. ...........
134 to 135
Phenolphthalein.. . . . . . . . .
(n.l.t.) 135*
Phenylsalicylate ...... " .
120 to 122
Phloroglucinol, R ..... " .
157
to 159
Pilocarpine nitrate .......
107 to 109
Procaine hydrochloride ...
111
to 113
Pyrogallol. ... ...........
114
to 116
Quinine ethylcarbonate....
(n.Lt.) 188
ResorcinoL ............
187 to 190
Saccharin..... ..........
127
to 129
Santonin .................
120 to 122
Scopolamine hydrobromide
235 to 237
Spermaceti. . . .. "
174 to 177
Suet, prepared. . . .. .
116
to 117
Sulfonethylmethane.... . ..
(n.Lt.) 78
Terpin hydrate ...........
Theophylline ............ '.
96 to 98
(n.Lt.) 183
Thymol. .. .. .. .. . .. . .. . ..
154 to 156
TrinitrophenoL ..........
142
to 146
Vanillin... . ............
(s.) 80
Wax (white aJ;)d yellow) ...
198 to 200
N.F.
52.5 to 53.5 Arecoline hydrobromide ...
34 to 40
Camphor, monobroma~d.
216 to 220
ChlorthymoL . . .
240 to 247.5 Cholesterol, R... . . .....
88 to 90
Chrysoidin, ba.se . ........ .
Cinchophen .............
123 (d.)
Cotarnine periodide .......
(a.) 28
Coumarin. . .. . . . . . . . . . . ..
(a.) 130
Dimethylaminoazobenzene,
(a.) 212(d.)
(a.) 115
198 to 201
36 to 42
41
to 43
(a.) 263
(n.l.t.) 74
30 to 35
50 to 57
38 to 54
195 to 197
(n.l.t.) 189
(a.) R about. (n.Lt.) = not le8s than.

.. Special method. s = sublimes.
173 to 177
(n.l.t.) 258
41
to 43
216
to 219
170 to 173
153 to 156
130 to 133
89 to 91
109 to 111
(n.l.t.) 222
169 to 171
190
to 192
42
to 50
45
to 50
74
to 76
115
to 117
269
to 272
48 to 51
121
to 123
80
to 82
62
to 65
169
74
59
146
213
142
67
to 171
to 76
to 61
to 148
117
to 216
to 144
to 69
R ..................... 115
to 117
p-Dimethylamino!lZobenzene, R......... .....

Ephedrine hydrochloride ..
Ethyl carbamate...
Guaiac............... . ..
Guaiacol carbonate. .. ...
Papaverine ...............
Phenylhydrazine
hydrochloride. R ......... ..
Pilocarpine hydrochloride ..
Salicin... . ...........
Sulfonmethane ...........
(d.) = with decomposition.
R.
73
214
48
85
86
145
to 75
to 220
to 50
to 90
to 88
to 147
240_
195
-l99
124
to
to
to
to
243
198
202
126
reagent .
231
4. What per cent error would be introduced in the determination of the

melting point of salicylic acid as found if uncorrected?
5. Given two samples which show the same melting points; how might
you determine whether or not they were the same substance?
Congealing Point.-The congealing or solidifying point is a

physical constant characteristic of certain liquids, such as oil
of anise, oil of eucalyptus, and the fatty acids of fixed oils,
which may serve as a criterion of purity and quality just as the
melting point of a solid does, since the presence of impurities
also causes a depression of the congealing point. The official
method for the determination of congealing points is as follows:
"Unless otherwise directed, about 10 cc. of the liquid to be
tested is introduced into a dry test tube of from 18 to 20 mm.
internal diameter, which is then cooled in water or a suitable
freezing mixture, the temperature of which should be about 5
lower than the supposed congealing point of the liquid. To
induce crystallization, rub the walls of the tube with a standard
thermometer, or add a small crystal of the substance being tested.
By alternate immersion of the tube in the bath, or removal from
the bath and constant stirring with the thermometer, the temperature is so adjusted that the greater part of the liquid gradually
congeals. The highest temperature remaining constant for a short
time during the congelation is defined as the congealing point."
The solidification temperature of fatty acids is commonly
referred to as the titer. The official procedure for the determination of the solidification of fatty acids is illustrated in the following exercise.
Exercise 76
Object.-Determination of the Solidification Temperature of

the Fatty Acids of Cottonseed Oil.
Materials Required.-50 cc. of cottonseed oil.
25 Gm. of potassium hydroxide.
100 cc. of glycerin.
15 cc. of alcohol.
"A standard thermometer meeting the following specifications: The
thermometer must have a zero mark, graduations of 0.1 0. between 10 and
60C., and auxiliary reservoirs at the upper end and between the 0 and 10
marks. The cavity of the capillary tube between the 0 and 10 marks must
be at least 1 cm. below the 10 mark, which must be from 3 to 4 cm. above
232
the bulb, the total length of the thermometer being about 38 cm. The bulb
should be about 3 cm. long and 6 mm. in diameter, the scale should be
etched on, and the graduations clear-cut and distinct. The thermometer
should be made of the best thermometric glass and thoroughly annealed, so
that scale errors will not increase after continued heating."
Procedure.-l. "Heat 75 cc. of glycerin-potassium hydroxide solution
(made by dissolving 25 Gm. of potassium hydroxide in 100 cc. of glycerin)
to 150C. in an 800-cc. beaker and add 50 cc. of the clarified fat, melted if
necessary. Heat the mixture for fifteen minutes with frequent stirring, but
do not allow the temperature to rise above 1500. When saponification is
complete, the mixture is homogeneous, with no particles clinging to the
beaker at the meniscus."
The oil is saponified by the KOR with the formation of the

potassium salts of the fatty acids and glycerin. See Saponification Value (page 350) for the reactions that occur.
2. "Pour the soap intq 500 cc. of nearly boiling distilled water in an
800-cc. beaker or casserole, add slowly 50 cc. of dilute sulfuric acid (made
by adding 1 volume of sulfuric acid to 3 volumes of distilled water), and
heat the solution with frequent stirring, until the fatty acids separate
cleanly as a transparent layer. Wash the acids with boiling water until free
from sulfuric acid, collect them in a small beaker and place on a boiling
water bath or steam bath until the water has settled and the fatty acids are
clear. Allow the acids to cool, melt and filter into a dry beaker while hot,
and dry for twenty minutes at 1000.
"Place 3 cc. of the dry acids in a test tube and add 15 cd. of alcohol. Heat
the solution to boiling and add an equal volume of ammonia water. A clear
solution should result."
The fatty acids are liberated from their potassium soaps by

the sulfuric acid. They are washed to remove potassium sulfate,
excess sulfuric acid, and glycerin and dried to remove the water.
If saponification is incomplete, the ,acids will be contaminated
with unsaponified oil which would alter the congealing point of
the acids. A turbidity in the alcohol solution indicates incomplete saponificat~on since the oil is insoluble in the diluted alcohol
and the ammonium soap is soluble.
.
3. "Cool the dry, filtered acids to from 15 to 20 degrees above the expected
reading, and transfer to a glass tube 25 mm. in diameter and 100 mm. in
length, the glass bellig 1 mm. in thickness. By means of a perforated cork
fasten the tube in a wide-mouth bottle of clear glass, approximately 70 mm.
in diameter and 150 mm. in height. Suspend the standard thermometer
in the melted acids so that it will serve as a stirrer, cooling if necessary, and
stir the mass slowly until the mercury remains stationary for thirty seconds.
233
Then allow the thermometer to hang quietly, the bulb in the center of the
acids and observe the rise of the mercury column. The highest point to
which it rises is the Solidifying Point of the fatty acids."
When the melted acids solidify, the "latent heat of fusion" is

liberated, and a rise in temperature occurs which remains constant for a flhort time. The fatty acids give a distinct rise in
temperature when they solidify, whereas the fats and fatty oils
are usually characterized by the fact that they yield a constant
temperature which in some cases is variable when they solidify.
1. The U.S.P. requires that the congealing point of stearic acid should not
be below MOC. Why?
2. The determination of the congealing point of oil of eucalyptus constitutes the official assay process. How does this determination serve as a
measure of the purity of the oil?
TABLE XXXII.--0FFICIAL SUBSTANCES WITH CONGEALING POINT
REQUIREMENTS
Substance
U.S.P.
Acid, glacial acetic ........................ .
Acid, oleic. . . . . . . . . . . . . .. ............. "
Acid, stearic ............................. .
Benzene, R .............................. .
Eucalyptol. .............................. .
Fatty acids from
oil, almond, expressed ................... .
oil, cottonseed. . . . . .. . ................ .
oil, olive. . . . . . . . . . . . . . . . . . . . . . . . .. . .. .
oil, theobroma. . . . . . . . . . . . . . . . .. . ..... .
soap, hard ........................... .
sodium stearate. . . . . . . . . . . .. . ......... .
zinc stearate. . . . . . . . . . . .. . ........... .
Oil of anise .............................. .
Oil of eucalyptl,ls. . . . .
. ........... .
Oil of fennel. . . . . . . ............ .
Paraldehyde. . . . . . . . . . . . . . . . . .. . ...... .
Phenol. ................................. .
Phenylhydrazine, R ....................... .
Suet, prepared ........................... .
Congealing
Point, cC.
(n.l.t.) 145
(n.m.t.) 10
(n.l.t.) 54
(n.l.t.) 5.2
(n.l.t.) 0
9 to 12
28 to 35
17 to 26
45 to 50
18 to 23
(n.l.t.) 54
(n.l.t.) 54
(n.l.t.) 15
(n.l.t.) 15.4
(n.l.t.) 3
(n.l.t.) 11
(n.l.t.) 39
(n.l.t.) 16
37 to 40
N.F.
Anethol. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 20 to 21
o-Toluidine, R ............................ (n.m.t.) 20
(n.m.t.) = not more than. (n.l.t.) = not less than. R. = reagent.
234
Boiling and Distilling Points.-One of the characteristic

properties of a liquid is its boiling point. If a substance consists
of a single compound, it is generally considered t,o be pure when
it distils at a constant boiling point, under cOIlstant' pressure.
In like manner, the purity of a mixture of substances, such as
a volatile oil, may be judged in part by the range of temperature
at which the substance distils.
FIG. 27.-Apparatus for boiling and distilling point determinations,
The boiling point of any liquid may be defined as that temperature at which the vapor pressure of the liquid is equal to the atmospheric pressure. Where a given temperature is stated as the
boiling point of a liquid, the inference is that the corresponding
pressure is standard atmospheric pressure, 760 mm., unless
otherwise given.
In the Pharmacopoeia, the boiling point is defined as that range
of temperature within which at least 95 per cent, by volume, of
the substance distils when treated in a definite manner.
The apparatus to be used (Fig. 27) and its construction are as
follows:
MELTING, CONGEALING, AND BOILING POIN;rS
235
A distilling bulb of from 50 to 60 cc. capacity to the lower part

of the neck; the len~th of the neck should be from 10 to 12 cm.
and its internal diameter from 14 to 16 mm. The outlet tube
of from 4' to 5 mm. internal diameter should be attached to the
neck at approximately its mid-point, forming an angle of
70 to 75 deg. with the lower portion of the neck.
A straight glass condenser with a water jacket from 40 to 60 cm.
in length; the distance from the upper end of the water jacket to
the neck of the bulb being from 18 to 25 cm.
In order to avoid the necessity for an emergent stem correction,
it is advisable that an accurately standardized thermometer of the
Anschutz type be used. If such a thermometer is not available,
a thermometer used for melting points may be employed, applying, the emergent stem correction as described under "Melting
Points," the length of the emergent stem being measured, in
degrees, from the lower end of the stopper to the highest point
of the boiling temperature. When placed in position, the lower
end of the thermometer should be from 2 to 3.5 cm. below the
center of the orifice of the outlet tube.
An asbestos board of from 12 to 15 cm. square and from 3 to 5
mm. in thickness, having a circular perforation, located centrally,
for the reception of the bulb. The edge of the asbestos around
the perforation should fit closely to the bulb when the latter is
set into it. The size of the perforation should be such that when
the bulb is set into it, the portion of the bulb below the upper
surface of ,the asbestos will have a capacity of from 3 to 4 cc.
Exercise 77
Object.-To Determine the Boiling Point of Carbon Tetrachloride.

Materials Required.-25 cc. of carbon tetrachloride.
A distillation apparatus as illustrated.
Procedure.-Place the asbestos" board on a tripod or other suitable
support. Introduce into the distilling bulb 25 cc. of the liquid to be tested,
insert the thermometer, stand the bulb in an upright position in the perforation of the asbestos board, and connect it with the condenser. Then
distil the liquid at the rate of 1 cc. for each 15 to 20 sec., noting the temperature as Boon as 5 drops of the liquid have distilled into the receiver and when
the last liquid evaporates from the bottom of the flask or when the specified
percentage has distilled over. Correct the reading for any variation in the
236
barometric pressure from the normal (760 mm.) by allowing O.lC. for each
2.7 mm., adding if the pressure is lower, and subtracting if higher than
760 mm.
The temperature obtained when a thermometer is immersed

in a boiling liquid is usually greater than the true boiling point
of the liquid, because the vapor bubbles form below the surface
of the liquid and consequently are at a pressure greater than
atmospheric. There is also an appreciable error due to superheating of the liquid, in this case. When the thermometer is
suspended in the vapor of the liquid, however, the bulb becomes
coated with a thin film of liquid in contact with its vapor, and
superheating is avoided.
It is often best to run a preliminary distillation, of a 25 cc.
portion of liquid, prior to the boiling point determination, in
order to ascertain the regulation of heating necessary to cause
the entire portion of volatile liquid to distil at the prescribed
rate. Superheating of the liquid and consequent bumping can
be prevented by placing a few small pieces of porous plate in the
distillation flask. The correction for the barometric pressure
may be made simply as illustrated in the following example:
If the range of temperature within which 95 per cent of glacial
acetic acid distils is 116.5 to 116.9C. at 733 mn1., the corrected
- 733) = 117.5, and
temperatures would be 116.5 + 0.1 ( 7602.7
60
116.9 + 0.le 2-; 733) = 117.9. The boiling point of the acid
should then be reported as b.p'133 = 117.5 to 117.9C. cor., the
subscript 733 indicating the barometric pressure at which the
determination was made, and the abbreviation" cor." indicating
that the boiling point as reported has been corrected to standard
pressure. If a long-stemmed thermometer is used in the determination, a further correction for the emergent stem must be
made, as explained under Melting Point, page 229.
A second method is given in the Pharmacopoeia for use with
Jiquids for which the permissible range in boiling temperature
exceeds 5C. The following is a description of the apparatus
required and the procedure to be followed:
A 200 cc. distilling bulb with an outlet tube at approximately
the center of the neck, forming an angle of from 70 to 75 deg.,
with the lower end of the neck. The length of the neck is from
237
10 to 12 cm. and its inside diameter is from 18 to 24 mm. The

length of the outle,t tube is from 10 to 12 cm. and its inside
diameter is from 5 to 6 mm.
A straight glass condenser, a thermometer, and an asbestos
board are required as in Method 1. The diameter of the perforation in the asbestos board is 25 mm.
Place the distilling bulb in an upright position in the perforation in the asbestos board and connect it with the condenser.
The outlet tube is to extend from 25 to 35 mm. into the condenser
beyond the connecting stopper.
Measure 100 cc. of the liquid to be tested, using a cylinder
having 1 cc. graduations. Note the temperature of the liquid and
transfer it as completely as possible to the distilling bulb. Use
this cylinder as the receiver for the distillate without rinsing out
any of the adhering liquid. Insert the thermometer through a
stopper in the position described in Method I and heat with a
Bunsen burner protected from drafts until the liquid begins
to boil, then distil at the rate of from 4 to 5 cc. per minute. The
temperature when 5 drops of the liquid have distilled into the
receiver is the minimum distillation temperature, and unless
otherwise specified, the maximum distillation temperature is the
temperature at which 95 per cent of the liquid has distilled over.
Bring the distillate to the same temperature at which the liquid
was originally measured, and note the volume. Correct the
telI\perature reading for barometric pressure and apply the emergent stem correction as described in Method I.
Liquids which begin to distil below 80C. are cooled to 10 or
15C. before measuring the 100 cc. for the test. The end of the
condensing tube is fitted with an adapter bent at a suitable angle
and the end of the adapter is passed through a cork inserted into
the receiving cylinder. The cork has a small perforation to
permit the exit of air. The receiving cylinder is kept immersed
in ice to within 2.5 cm. of it~ height during the distillation.
When boiling points are given with a range instead of with a
single figure, the figures are to be regarded as inclusive. The
boiling points of such substances as acetone, ether, ethyl oxide,
alcohol, glacial acetic acid, and tetrachlorethylene should be
regarded as informative rather than as criteria of identity and
purity since only approximate boiling points are given. Definite
requirements relative to the per cent of a substance which
238
should distil within a certain range of temperature and above or

below a certain temperature are given in the Pharmacopoeia and
National Formulary as follows: 85 per cent of creosote from
creosote carbonate distils at 200 to 220C., 90 per cent of creosote
distils at 203 to 220C., 90 per dmt of cresol distils at 195 to
205C., 90 per cent of oil of turpentine and of rectified oil of turpentine distils at 154 to 170C., and 85 per cent of liquid guaiacol
distils at 200 to 210C. It is required that the' volatile oil
separated from copaiba by steam distillation should not boil
below 250C., that less than 10 per cent of oil of dwarf pine needles
distils below 165C., and that the boiling point of liquefied phenol
should not rise above 182C. when distilled.
Those official substances for which a definite range of boiling
points is specified and within which range 95 per cent of the
substance should distil are given in the following table.
TABLE XXXIII.-OFFICIAL SUBSTANCES WITH BOILING-POINT RANGE
Substance
Boiling Point, cC.
U.S.P.
Amyl alcohol, R ........................ 128 to 132
Aniline, R ............................. )83 to 186
Benzene, R .............. ' .. .. ...... ... 79.5 to 81
Benzin, purified petroleum. . . . . . . . . . . . . .. 35 ~O 80
Butyl alcohol, R ........................ 116 tb 118
Carbon disulfide, R. . . . . . . . . . . . . . . . . . . .. 76 to 78
Carbon etrachloride. . . . . . . . . . . . . . . . . . . .. 76 to 78
Ethyl a tetate, R. . . . . . . . . . . . . . . . . . . . . .. 76 tb 77.5
Ethyl chcloride. ... . .. . . . . . . . . . . .. . . . . .. 12 to 13
Eucalyptol. . . . . . . . . . . . . . . . . . . . . . . . . . . .. 174 to 177
Eugenol ............................. " 250 to 255
Furfural, R, ........................... 150 to 152
Guaiacol, solid. . . . . . . . . . . . . . . . . . . . . . . . .. 204 to 206
Kerosene, R ............................ 180 to 300
Methanol, R.. ........................ 64 to 67
Methyl salicylate ....................... 219 to 224
Oil of mustard, volatile .... , ............. 148 to 154
Paraldehyde. . . . . . . . . . . . . . . . . . . . . . . . . . .. 120 to 125
Terebene .............................. 160 to 172
Toluene, R ............................. 110 to 111
Xylene, R .............................. 137 to 140
N.F.
Ethylacetate.......................... 75 to 77
Anethol. ... : ... . . . . . . . . . . . . . . . . . . . . . . . .. 234 to 237
R. = reagent.
239
1. Define the terms boiZi'ng point and distilling point.

2. How is the distillation range of the cresol and of the creosote in creosote
carbonate direc'ted to be determined in the Pharmacopoeia?
3. If ether contained alcohol as an impurity, would the boiling point of
the ether be raised or lowered?
4. A sample of carbon tetrachloride is found to distil between 76.8 and
77.90. at 664 mm. Calculate the corrected distilling range of the carbon
tetrachloride.
6. Consult the appended table and list those substances the boiling points
of which should be determined by method II of the U.S.P.
6. Which is more pure as indicated by the required boiling point ranges,
ethyl acetate of the N:F. or ethyl acetate reagent of the U.S.P.?
CHAPTER XIV
REFRACTOMETRIC MEASUREMENTS
The index of refraction is a physical constant frequently made

use of i.n the determination of the identity and purity of drug and
food products. In many cases, it may be used to determine
quantitatively the strength and purity of solutions or the proportions in which liquids are mixed; e.g., the percentage of sugar in
syrup can be estimated directly from the refractivity of the solution, and the percentage of alcohol in water can also be aetermined
in this way. Although the refractive index is a constant characteristic of many substances such as fats, fatty
oils, waxes, sugars, organic solvents, etc., it is
applied
almost exclusively in the official stand[
ards as a criterion of the purity of volatile oils.
Index of Refraction.- When a raJ;" of monochromatic light passes from one transparent
substance into another of different optical
density, it is deflected or refracted except
FIG. 28.-Refrac- when it enters perpendicularly to the surface
tion of light.
of con t act between the substances. The
extent and direction of the deflection are dependent upon the
difference between the densities of the two substances. The
angle between the ray in the first medium and a perpendicular to
the dividing surface is termed the angle of incidence i, and the
corresponding angle in the second medium is called the angle of
refraction r. The sin of i and the sin of r are directly proportional
to the velocities of light in the two media. The ratio sin i/sin r
is termed the index of refraction. Thus, the velocities of light
in air and in water are in the ratio of about 4 to 3. The index of
refraction of water with respect to air is therefore about 1.333.
In Fig. 28, I is the less dense medium and II the more dense
medium. A ray of light passing obliquely from I to II will be
deflected so that the angle of refraction r will be less than the
240
itEFRACTOME'l'RIC MEASUREMENTS
241
angle -of incidence i, and according to the law of refraction, the

relationship between 'Fhese two angles will be sin ilsin r = N In
where n is the ip.dex of refraction of the less dense and N the index
of refraction" of' the more dense medium. When the angle i is
incr~sed, the angle r also is increased and attains its maximum
value when the angle of incidence enters horizontally at 90 deg.,
tm"'med grazing incidence, or at a right angle. In the latter case,
since the sin pf 90 deg. = 1, -the above equation becomes 11sin
r = N In or sin r = niN. The angle 90 deg. is known as the
critical angle and the ray of incident light passing from one
mediu:qJ. to another at this angle is known as the critical ray.
If the angle r is increased to a value at which i becomes equal to
90 deg., the beam of light will no longer pass from the first
medium to the second but will travel through the first medium to
the dividing surface at an angle of 90 deg. with the perpendicular
to the surface. If r is smaller than this particular value, however,
the ray of light will pass on through the second medium, and, if
greater, all light will be reflected from the surface back into the
first medium. The critical ray is thus seen to make a boundary
between the reflected and refracted light. This furnishes the
basis for the observation of the refractive end point used in many
refractometers.
Refractometers.-Indices of refraction may be determined by
means of a polarizing microscope or refractometer. Refractometers are of several types. Some of these give figures on an
arbitrary scale, the refractive index being found by reference to a
table furnished with the instrument. The principal instruments
in use are (1) the Pulfrich refractometer, an instrument adapted
for use with liquids and solids of all degrees of refractive power,
used chiefly for purely scientific investigations; (2) the Zeiss
butyro-refractometer, designed especially for the examination
of fixed oils and fats; (3) the Zeiss immersion refractometer,
especially constructed for the examination of aqueous, oily, or
alcoholic liquids. when a large ;mount of sample is available; (4)
the Abbe refractometer, an instrument designed for the examimi,tion of liquids, solids, and plastic bodies. Since this last instrument is best suited for the examination of all of the official substances for which refractive indices are given, it is described in
considerable detail.
242
The Abbe Refractometer.-Because of its simplicity, ease of

manipulation, and the small amount of material required in its
use, the Abbe refractometer is extensively used in pharmaceutical
analysis. The wide range of indices measurable with this instrument and the fact that it can be used with ordinary white light
make it capable of more generaL application than any of the
other refractometers mentioned above. When measuring the
refractive index of liquids, only a few drops, sufficient to form a
FIG.
29.- Abbe refractometer.
film, need be placed between the prisms of the instrument.

This is a distinct advantage over the other forms of refractometers, for frequently only a small quantity of the substance to
be examined is availabl~, and if the liquid is colored or turbid,
the thinness of the film permits a determination of the index of
refraction without difficulty.
The Abbe refractometer (Figs. 29 and 30) consists of four
essential parts, namely: (1) a telescope E provided with an
objective, an eyepiece, and a cross-hair disk. The purpose of the
telescope is to form an image of the border line of total reflection
243
at the intersection of the cross-hairs where the image may be

observed. (2) Two similar glass prisms (Abbe prisms) of high
refrac~ive index cemented into hollow, water-jacketed, mounting
cases, so constructed that when temperature control is required,
water can be circulated about the prisms. The exposed surfaces
of the upper prism B are highly
polished, while the hypotenuse surface of the lower prism A, which
serves solely for the purpose of illumination, is finely ground. The lower
prism mount is hinged to the upper
prism mount and the latter is rigidly
attached to the index arm or alidade.
When the two prisms are clamped
together, a space of from 0.1 to 0.15
mm. separates their surfaces. (3) The
sector H is a metal arm to which the
telescope is rigidly attached. One
end of the sector is attached to the
upright of the base in such a manner
that the whole sector can be rotated
about an axis, which coincides with
the axis of rotation of the Abbe
prism, so that various conditions of
illumination can be accommodated.
On the upper end of the sector a scale
G is mounted which is graduated
directly in terms of refractive index FIG. 30.- Path of light, etc.,
through Abbe refractometer.
of the D line (sodium light) at 20C.
An alidade or movable arm J rests against the sector, but it
can be rotated as a unit with the Abbe prism independently
of the sector. The upper end of the alidade bears an index
mark which moves along the edge of the sector scale when
the arm is rotated. A magnifier F is attached to the alidade
in most instruments to facilitate the reading of the index
scale. A screw M is used to make fine adjustments of the
border line at the cross-hairs. (4) Compensating prisms are
used to correct for the dispersion of light. Thus, when white
light is used in the place of sodium light, the beam is split
244
up into colored beams, since short wave lengths of light are

refracted to a greater extent than are lopg wave lengths.
Unless this dispersion is compensated -for in some way, the
border line becomes a broad, varicolored blind which cannot
be brought into accurate coincidence with the, cross-hairs of the
telescope. The compensator functions to bring the various wave
lengths of light together again. It is placed hetween the upper
prism and the eyepiece and is rotated by me!\ns of the milled
head D.
FIG. 31.-Temperature control apparatus for Abbe refractome'ter. W.

Water supply. WR. Water-pressure regulator. S. Spiral heater. B. Bunsen
burner. C. Water-pressure regulator. P. Inlet stopcock. R. Refra~tometer.
Regulation of Temperature.-Since the density of botth the

prisms and the substance under investigation varies witl;t the
temper~ture, it is necessary in refractivity measurements or\ any
degree of accuracy to be able to maintain the prism and, the
substance at constant temperature. In the Abbe refractome~er,
this control of temperature is accomplished by circulating witter
from a thermostat or other suitable constant temperature device
about the jacketed prisms, the water entering the inlets I and'L
passing out 9f the outlets 0, the temperature at which the measurement is made being observed on the thermometer T. A
temperature control device is illustrated in Fig. 31.
245
Exercise 78
Object.-To Determine the Refractive Index of Oil of Orange.

Materials Required.-About 1 cc. of oil of orange.
A pipette.
An Abbe refractometer.
Procedure.-l. ~tate the prisms and telescope so that the lower prism
may be swung opelJ easily. Attach the test plate to the upper prism, by
means of the monobrom-naphthalene or other liquid provided with the test
plate, by moistening the test plate with the liquid and pressing it against
the upper prism.a Focus the light by means of the lower prism, on the
test plate, rotate the prism by means of the alidade until the border line
of total reflection coincides with the cross-hairs in the telescope, and take
the reading of the,refractive index.
The refractiye index of the test plate is etched upon it, and the
reading of th~instrument should correspond to that given on this
plate. If thtJ instrument is not in adjustment, a correction should
be applied to the readings obtained.
2. Remo'l{e the test plate, clean the upper prism with cotton saturated
with alcohol, clamp the prisms together loosely, introduce 2 or 3 drops of
the volatilll oil into the groove at the side of the prisms, and clamp the
prisms together firmly. Adjust the mirror K so that the light is reflected
upon the lower prism, and rotate the alidade until the border line between
the lightl and dark halves of the field of view exactly coincides with the
cross-hai,'rs of the telescope, rotating the compensator prisms to obtain a
sharp uncolored border line if necessary. Read the refractive index of
the oil directly from the graduated sector scale to the fourth decimal place.
Move the aJidade and again determine the refractive index until three
readings are obtained, taking the mean of the readings as the refractive
index of the oiL Note the temperature at which the refractive index is read.
The'readings on the graduated scale of the compensator drum

may be referred to tables of mean dispersion and conversion
sup~lied with the instrument.
The refractive index of the oil
sho.uld be expressed as, e.g., N';oo = 1.4725 where N~oo denotes
ths index of refraction for the D line (sodium light) measured
at 20C.
1. Define the terms index of refraction, critical ray, and angle of incidence.
2. What advantages does the Abbe refractometer possess over the other
types of refractometers?
246
3. A range is given for the refractive index of most of the official volatile
oils. Why?
4. How do variations in temperature affect the inqices of refraction of
liquids?
6. Would you expect that oil of orange would have a greater or smaller
index of refraction at 25 than at 20C.? Why?
TABLE
XXXIV.-OFFICIAL
SUBSTANCES
WITH
I
RE~UlRED
REFRACTIVE
INDICES
Substance
U.S.P.
Eucalyptol. ......................
Oil of anise .......................
Oil of bitter almond ...............
Oil of chenopodium ...............
Oil of cinnamon ...................
Oil of clove .......................
Oil of coriander ...................
Oi~ of dwarf pine needles ...........
Oil of eucalyptus ..................
Oil of fennel. .....................
Oil of juniper .....................
Oil of lavender ....................
Oil of lemon ......................
Oil of mustard, volatile ............
Oil of myristica ......... .........
Oil of orange .....................
Oil of peppermint .................
Oil of rose ........................
Oil of rosemary ...................
Oil of santal. .....................
Oil of sassafras ...................
Oil of spearmint ..................
Oil of theobroma ..................
Oil of turpentine ..... , ............
N.F.
Anethol. .........................
Oil of caraway ....................
Temperature,
Official requirement
C.
20
20
20
20
20
20
20
20
20
20
20
30
20
20
20
20
40
20
1.455 to 1.460
1. 5530 to 1. 5600
~.5428to 1.5439
1. 4723 to 1. 4790
1 .'6020 to 1. 6135
1. 5300 to l. 5350
1.4620 to 1. 4720
1. 4580 to 1. 4700
1 . 4600 to 1. 4690
1 . 5280 to 1. 5380
1. 4780 to 1. 4840
1. 4590 to 1. 4700
1.4742 to 1.4755
1 . 5268 td 1. 5280
1. 4740 to'!. 4880
l.4723 to'1.4737
l. 4600 to'!. 4710
1.457 toh.463
1. 4640 to b. .4760
1.5000 to 1.5100
1 . 5250 to 1. 5350
1. 4820 to 1'.4900
1 . 4537 to 1. 4578
1. 4680 to 1. ~ 780
25
20
1. 5580 to 1. 5~10
1.484 to 1.488
20
20
20
20
20
20
CHAPTER XV
ROTATORY POWER
Many organic substances, such as certain constituents of the
volatile oils, alkaloids, and sugars, possess the power of rotating
the plane of polarized light when the latter is passed through
solutions containing them. Substances which possess this power
are said to be optically active and are designated as dextrorotatory when the direction of the rotation is toward the right and as
laevorotatory when it is toward the left.
The optical activity of many substances is a function of their
chemical constitution as well as of their concentration. Consequently, a determination of the rotatory power or lack of
rotatory power of a substance may serve as a means of establishing both its identity and its purity. In some cases, the me asu~ement of the optical activity of a substance may also give
some indication of its therapeutic value; e.g., the laevorotatory
alkaloid hyoscyamine is much more active as a mydriatic than
its optically inac.tive isomer atropine.
In a ray of ordinary light, the vibrations are transverse, i.e.,
they take place in a. plane at right angles to the direction of
propagation, but the vibration direction is constantly changing.
In a ray of plane-polarized light, commonly termed polarized light,
the vibrations are also transverse, but they take place in only
one direction.
The polariscope and the polarizing microscope are used in the
determination of the optical activities of liquid and solid substances, the polarizing microscope being employed more often
for the examination of solid SUbstances, while the polariscope
is used primarily for the examination of liquids.
When a ray of ordinary light is passed through a specially
constructed prism made of Iceland spar, such as the Nicol's
prism, the emergent light wave vibrations take place in one
plane, and the light is' said to be plane-polarized. If another
247
248
Nicol's prism is used to examine the plane-polarized light, it will

be found that on rotating the latter prism, the field of view
appears alternately light and dark, the minimum brightness
following the maximum as the prism is rotated through an angle
of 90 deg. The prism by means of which the light is polarized
is termed the polarizer, and the second prism, by means of which
the light is examined, is called the analyzer.
If, when the field of view appears dark, as when the axes of the
two prisms are at right angles to each other, a tube similar to
that shown in Fig. 32 or 33 is filled with an optically active
liquid and placed between the polarizer and analyzer, the ~ld
will become light due to the rotation of the plane of polarization
FIG. 32.-Polariscope tube.
FIG. 33.-Polarisoope tube

with jacket and thermometer
well for constant temperature
control.
by the optically active substance. The extent to whichlthe plane

of polarization has been rotated can be determin~d b~ turning
the analyzer through a certain angle until the field becomes dark
again. When it is necessary to turn the analyzer to 'the right
(clockwise), the optically aptive substance is said to be dextrorotatory; when the analyzer must be turned to the left (counterclockwise), it is said to be laevorotatory.
It is possible to obtain a position at which the field of view
becomes dark by rotation of the analyzer to either the right or
left, since in one complete rotation of the prism through 360 deg.,
there are two positions 'of the analyzer, 180 deg. apart, at which
the field becomes dark, and, likewise, two positions at which
there is a maximum of light. In determining the sign of activity'
of a given substance, it is customary to take the direction in which
the "rotation required to reproduce the dark field is le,ss than
90 deg.
The angle of rotation of the polarized light on passage through
a substance varies directly as the length of the column of sub-
ROTATORY POWER
249
stance through which the light passes. In homogeneous mixtures

or solutions, the angle of rotation depends upon the concentration
of the optically active substances present. When more than one
optically active substance is present in a solution, the resultant
rotation will be the same as the sum of the several rotations of
each separate constituent in the same volume of solution, provided that the substances do not react with each other. The
angle of rotation also varies with the wave length of the light
employed (the shorter the wave length the greater the angle of
rotation), with the temperature, and with the nature of the
substance. When all, of these factors are taken into account, the
results may be calculated in terms of specific rotation or specific
rotatory power.
The specific rotation of an optically active liquid 'is defined
as the angle of rotation in degrees through which the plane of polarization of monochromatic (D) light is rotated by passage through a
liquid containing 1 Gm. of active substance in a volume of 1 cc.
determined in a tube having a length of 1 demo Since the specific
gravity of liquids as well as the optical activity of many substances is influenced by the temperature, it is necessary to indicate
the temperature at which the rotation and the specific gravity are
determined. The temperature specified in the two official
standards for the measurement of optical activity is generally
250., except in the case of sugar which is determined at 200.
by international agreement.
The calculation of the specific rotatory power of an optically
active liquid or solution of an optically active solid may be made
from the following general formulae:
1. For liquid substances [alb =
II. For solutlons [alb
lOOa
lpd
or [alb
l~a
ia
For calculating the specific rotation [k] using these formulas,

the determination of the following factors is necessary:
a = observed rotation in degrees of the liquid at temperature
t using a sodium light.
1 = the length of the tube in decimeters.
250
the specific gravity of the liquid or solution at the temperature of observation.

p = concentration of solution expressed as the number of grams
of active substance in 100 Gm. of solution.
c = concentration of solution expressed as the number of
grams of active substance in 100 cc. of solution.
In expressing the specific rotation of a substance in solution
the concentration and solvent used must always be stated. The
temperature should also be recorded; e.g., [al~Oo = -38 means
that the specific rotation of the substance determined at 20C.
using sodium light is laevorotatory 38 deg., and [al~Oo = +16
means that the substance is dextrorotatory 16 deg. when the
Elpecific rotation is determined at 20C., using sodium light.
Polarimeters.-The instruments used to measure the optical
activity of liquid substances are called polariscopes or polarimeters
=
,,~F
r
l\'
,. . ----------.1
lI/E ISID,~-=,=o;::::==:i~,lc ISIB
AO,~R{
~1f,"!.A
ClW
FIG. 34.-Diagram of the optical parts of a polarimeter.
and saccharimeters. Polarimeters permit the angle of rotation to

be read off in degrees or fractions of a degree of a circl y, while
saccharimeters are usually constructed and set to give readings
directly in terms of percentage of sugar. The so-called halfshadow instruments known as the Laurent and the Schmidt and
H ansch polarimeters are most generally used in pharmaceutical
work.
In the polarimeter of Laurent and that of Schmidt and Hansch,
the optical parts may be represented diagrammatically as in Fig.
34. Monochromatic light (sodium light) from the source L,
passes through the lens A, which renders the rays of light parallel,
and then passes through the polarizing prism B, which renders
the light ray plane-polarized. It then passes through the observation tube 0, which contains the substance being examined,
and thence through the analyzer prism D. The field of view is
observed through the telescope FE. The circular opening
of the tube carrying the polarizer is one-half covered by a thin
quartz plate as illustrated in 0', the thickness of which is so
adjusted that the light in passing through the plate is altered
ROTATORY POWER
251
in phase by half a wave length but remains plane-polarized. In

this way, two beams of polarized light are produced. If the
polarizer is rotated so that the plane of polarization forms an
angle X with the quartz plate, the planes of polarization will
also be inclined at an angle 2X. This is termed the half-shadow
angle. On rotating the analyzer, a position will be found at
which one beam of light will be completely, and the other only
partly, extinguished. One half of the field of' view will then
appear dark while the other half will remain light. Upon rotating
the analyzer farther through the angle 2X, a second position will
FIG. 35.-A polarimeter.
be found at which the second beam of light will be extinguished

while the first beam becomes visible. At this position of the
analyzer, that half of the field which formerly appeared dark will
be light and formerly light will be dark. When the analyzer
occupies an intermediate position, a field of view of uniform
brightness will appear, and this is the position at which the
analyzer should be set.
The complete polarimeter is shown in Fig. 35. At the end S,
which'is directed toward the soutee of illumination, is the lens.
The polarizing prism is at P, and it is connected with a lever
L, by means of which it can be rotated to alter the angle of halfshadow. The observation tube is placed in the central part
of the instrument and protected from extraneous light by a
hinged cover. The analyzer is placed at A, and it can be rotated,
252
independently of the graduated circle, by means of a screw. This

permits the correction of the zero point of the instrument'. F
is a telescope with an eyepiece by means of which the field of
view is observed. K is a graduated disk which can be made to
rotate with the analyzer and telescope past the fixed verniers n
on the graduated circle. Most instru~ents are provided with
one or more magnifying lenses to increase the accuracy of
readings .
.The official polarimetric determinations should be carried out
in a dark room using monochromatic light. Monochromatic
light can be obtained easily by introducing a fused bead of sodium
chloride on a platinum wire or gauze into the non-luminous flame
of a gas burner.
The observation tube, in which the liquid to be examined is
placed, usually consists of a tube of thick glass with accurately
ground ends, closed by circular glass plates with parallel sides
which are pressed against the ends of the tube by means of screw
caps. Other polarimeter tubes are constructed with a jacket
through which water may be circulated to give a constant temperature. Since the unit of length in polarimetric me/asurements is 1
dcm., the observation tubes are generally of 1 dcm. length or some
multiple or fraction thereof, e.g., 2.0, 0.5, and 0.'25 d~. The
length of the tube used in any defin)te determination usually
depends upon the depth of color in the liquid and the magnitude
of its rotatory power.
Readings made on a saccharimeter may be converted into the
angular degrees of a polarimeter by means of the following factors:
1 division of the Laurent sugar scale = 0.2167 deg. angular
rotation D.
1 division of the Ventzke or Schmidt and Hansch sugar scale
= 0.3468 deg. angular rotation D.
Exercise 79
Object.-To Determine the Specific Rotation of Sucrose.

Materials Required.-26 Gm. of cane sugar.
A 100 cc. volumetric flask calibrated at 20C.
A polarimete'r with a 200 mm. observation tube.
Procedure.-l. Adjust and determine the zero point of the polarimeter.
ROTATORY POWER
253
The polarimeter should be set up so that the polarizer end is in

direct line with the brigp.t sodium flame and about 4 to 6 in. from
it. Place a 200 mm. observation tube filled with distilled water
in the tube holder between the polarizer and the analyzer and
focus the telescope eyepiece on the line bisecting the field of
view, rotating the analyzer if necessary in Qrder to obtain unequal
illumination of the two halves of the field. Now determine the
zero point by rotating the analyzer until equal illumination of the
two halves of the field is obtained. This position should be
approached several times from either side of the zero point,
readings being made from the verniers, and the mean of at least
four readings taken as the zero point. Since the zero point is
altered by the alteration of the angle of half-shadow, the lever
which rotates the polarizer must be set before the zero point is
determined. It is possible to rotate the analyzer in some instruments without rotating the graduated disk, and it is possible,
in such cases, to adjust the instrument so that the zero point
correction is eliminated.
2. Dissolve 26 Gm. of sucrose, accurately weighed and previously dried
to constant weight at 105C. in sufficient distilled water to make 100 cc. at
20C.
The sugar is dried to drive off moisture. The solution should

be made up in a volumetric Ilask calibrated at 20C., and the
final adjustment of the volume should be made at the same
temperature.
3. Fill a clean dry observation tube with the solution. Determine the
value of the angle of rotation. Calculate the specific rotation of the sample
examined.
A polarimeter tube may be cleaned and dried easily by washing

it successively with water, alcohol, and ether. IIi the determination of the angle of rotation, a number of successive readings
should be taken as described under procedure 1, and their mean
taken as the angle of rotation. After correcting for the zero
point of the instrument, the specific rotation can be calculated
by the formulae given on page 249.
The specific rotation of the official sucrose must not be less
than +65.9 deg. when determined by the above method.' Com-
254
pare the specific rotation of the sample examined with

requirement.
th~
official

1. What is polarized light? How can it be obtained?
2. Show diagrammatically the essential parts of the polariscope used
and explain the function of each part.
3. What factors influence the rotatory power of a substance?
4. Define the term specific rotatory power.
TABLE XXXV.-OFFICIAL SUBSTANCES WITH REQUIRED ANGULAR ROTATION
Official Requirement
= Angular rotation
Substance
U.S.P.
Copaiba, volatile oil from ....
Methyl salicylate from gaultheria ....................
Oil of anise .................
Oil of bitter almond. . . .. . ..
Oil of cinnamon. . . . . . . . . .. .
Oil of clove. . . . . . . . . . . . .
Oil of coriander. . . . . . . . . . ..
Oil of fennel. ...............
Oil of juniper. . . . . . . . . . . . . ..
Oil of lavender ..............
Oil of lemon. . . . . . . . . . . . . ..
Oil of dwarf pine needles .....
Oil of myristic a . . . . . . . . . . . ..
Oil of orange .... ,. . . .. . ....
Oil of peppermint ...........
Oil of rose ..................
Oil of rosemary .............
Oil of santal. ...............
Oil of sassafras ..............
Oil of spearmint .............
Terebene. . . . . . . . . . . . . . . . . ..
not less than _7 0
a = not more than -1.5 0

a = +1 to _20
a = inactive to +0 0 10'
a = +1 to _1 0
a = not more than _10 10'
a = +8 to + 15
a = +12 to +24 0
a = 0 to -15..0
a = -3 to -10 0
a = +57 to +65.6 0
a = -5 to _120
a = + 10 to +30 0
a = +94 to +99 0
a
-23 to -32 0
a = 1 to 40
a = 5 to +10 0
a = -15 to -20 0
a = +2 to +4 0
a = -48 to -59 0
a = not more than +0.3 0
N.F.
Anethol. . . . . . ... . . . . . . . . . . ..
Oil of bergamot .............
Oil of bitter orange. . . . . . . . ..
Oil of caraway ..............
Oil of cardamom ............
Oil of myrcia ................
Oil of orange flowers. . . . . . . ..
Oil of pimenta ..............
Oil of thyme .... , ...........
Salicin. . . . . . . . . . . . . . . . . . . ..
a = 0 to +0.08 0
a = +8 to +240
a = +88 to +980
a = +70 to +80 0
a = +22 to +440
a = not more than _3 0
a = + 10 3~' to +9 0 8'
a = 0 to _4
a = not more than 4 0
a = 62 to 67 0
255
ROTATORY POWER
5. Explain what the following terms mean: (1) [aut = +4.210; (2)
= -10; (3) dextrorotatory; (4) optically inactive.
6. The optical rotation of a sample of oil of chenopodium determined in a
100 mm. tube and having a density of 0.9750 is found to be -12.25 deg.
at 21. Calculate the specific rotation of the oil.
7. A sblution containing 10 Gm. of sugar in 100 cc. was found to exhibit
an angular rotation of +11.5 deg. when determined in a 100 mm. observation tube. Calculate the specific rotation of the sucrose.
[aUf
The official substances with their optical activ.ity requirements

may be classified as follows:
1. Those which are required to be optically inactive.
U.S.P. Atropine, eucalyptol, eugenol, methyl salicylate
(synthetic and from Betula), vanillin, and volatile oil of mustard.
N.F. Anethol, arecoline hydrobromide, and papaverine
hydrochloride.
2. Those which are required to be laevorotatory.
U.S.P. Cocaine, codeine, colchicine, honey, menthol,
quinine, quinine bisulfate, quinine sulfate, and santonin.
N.F. Cinchonidine sulfate.
TABLE XXXVI.-OFFICIAL SUBSTANCES WITH REQUIRED SPECIFIC ROTATION
Substance
U.S.P.
Camphor" natural
Dextrose .........
Ephedrine hydrochloride ........
Ephedrine sulfate ..
Epinephrine ......
Oil, chaulmoogra ..
Ethyl chaulmoograte ...........
Lactose ...........
Scopolamine hydrobromide .....
Sucrose ..........
N.F.
Ephedrine sulfate,
solution of ......
Solvent
used
Concentrationin
.100 cc.
[alD =
[aut =
specific rotation
Alcohol
Water
10 Gm.
10 Gm.
[aUf = +52.5 to +53
Water
Water
0.5 N HCI
Chloroform
5Gm.
5Gm.
5 Gm.
10 Gm.
[aut
[aut
[aut
[alit
Chloroform
Water
50 cc.
[a]i,s = not less than +44.5
10 Om. [aUf = +52.2 to +52.5
Water
Water
5Gm. [aut = -22 to -25.75

26 Gm. [alt"O = not less than +65.9
Water
3 Gm.
=
=
=
=
+41 to +42
-33 to -35.5
-29.5 to -32 0
-50 to -53.5
+48 to +60
[a11&0 = -28 to -30
256
3. Those which are required to be dextrorotatory.

U.S.P. Quinidine sulfate.
N.F. Cinchonine sulfate.
4. Those for which the angular rotation at 25C. determined
in a 100 mm. observation tube is given in Table XXXV, page 254.
5. Those for which the specific rotation is given in Table
XXXVI, page 255.
All of the o:ffi~ial determinations of specific rotation are performed in a 200 mm. tube, except that of scopolamine hydrobromide in which a 100 mm. tube is used.
The measurement of rotatory power is made the basis for the
assay for dextrose in the ampuls of dextrose, ampuls of dextrose
and sodium chloride, and isotonic solution of dextrose and
sodium chloride of the N.F.
CHAPTER XVI
VISCOSITY MEASUREMENTS
The viscosity of a fluid substance is constant for any given

temperature and is a measurable characteristic of the substance.
It is used chiefly as an index of the composition and lubricating value of oils. The viscosities of solutions and liquid
mixtures often vary with their concentration and composition,
however, this property may be used, in many cases, as a rapid
means of analysis. The U.S.P. utilizes the viscosity as a means
of standardizing liquid petrolatum which is employed therapeutically as an intestinal lubricant.
In the U.S.P., viscosity is defined as "a term used to denote
the relative degree of fluidity of a liquid." This definition is
confusing, since fluidity is the reciprocal of viscosity; i.e., fluidity
= l/viscosity. Fluidity may be regarded as a measure of the
tendency of a liquid to flow, whereas viscosity is a measure of the
resistance which a liquid exerts against the tendency to flow.
Viscosity may be defined as the force of friction which tends
to retard movement in a fluid body. The unit of absolute viscosity
is defined as the tangential force required to move a unit area of
plane surface within the liquid with unit velocity relative to another
parallel unit area of plane surface one unit distant from the former
surface, where the unit of force is the dyne (the force necessary
to produce an acceleration of 1 cm. per second per second on a mass
of 1 Gm.), the unit of velocity is the centimeter per second, the unit
of area is the square centimeter, and the unit of distance is the centimeter. This unit of absolute viscosity is known as the poise and
it is generally expressed as dyne-second per square centimeter.
The poise is a relatively large unit, and the one-hundredth part
of ~he poise, or a centipoise, is commonly employed as the unit of
absolute viscosity.
Instead of giving results in terms of absolute viscosity, most
methods of determination give the relative viscosity; i.e., the
257
258
viscosity of the liquid compared to that of some standard of

known viscosity, such as water under similar conditions. Since
the relative viscosities obtained by determinations with different
types of apparatus are not the same, the kinematic viscosity,
which is the ratio of the absolute viscosity in poises to the density,
has been adopted in the U.S.P. as the method of expressing
viscosity. As the viscosity of a liquid changes very rapidly
with the temperature, decreasing about 2 per cent for each
degree rise in temperature, it is important to know the exact
temperature of the liquid at the time of measurement and to
use an instrument provided with some means whereby the sample
under examination may be maintained at constant temperature.
Apparatus.-In the official determination either the Eng I e r 0 r
Saybolt viscosimeter may be employed, and the results obtained
with these instruments may be converted into the kinematic viscosity
by the use of a conversion table.
Both of these instruments are constructed to measure the tim e
required for a certaiJ). amount of
fluid to pass through a capillary
tube. When the Engler instrument
is used, the viscosity is expressed in
terms of "Engler degrees," obtained
FIG. 36.-Saybolt universal vis- by dividing the time of discharge
cosimeter.
for the liquid being tested by the
time of discharge of the same volume of water at 200. In this
country, the Saybolt universal viscosimeter is most commonly
employed in the determination of the viscosity of lubricating
oils. The results obtained with this instrument are expressed
as the time in seconds required for the discharge of the liquid
being tested.
The Saybolt universal viscosimeter (Fig. 36) consists of a metallic
cup of about 80 cc. capacity to contain the liquid under examination with an outlet in the bottom closed by a cork stopper.
This cup is surrounded by a jacket which serves as a constant
temperature bath. Most instruments are provided with devices
VISCOSITY MEASUREMENTS
259
for stirring and heating the bath, with a thermometer to measure

the temperature, and with a receiving flask graduated to contain
60 cc. at 20C.; a stop watch with which the time of discharge
of the liquid may be measured is usually a part of the standard
equipment supplied with the instrument, but any stop watch
may be used.
Exercise 80
Object.-To Determine the Kinematic Viscosity of Liquid

Petrolatum. '
Materials Required.-A Saybolt viscosimeter.
A stop watch.
100 cc. of liquid petrolatum.
Procedure.-l. Assemble the viscosimeter, fill the constant temperature bath with water, and bring the temperature of the latter to about 4QC.
Tightly stopper the outlet tube, and pour the sample of petrolatum into
the oil cup until the level is above the rim of the main cup. When the
temperature of the oil sample is 37.8C., pipette the excess oil from the
overflow cup until the level in it is below the rim of the main cup.
The water in the temperature bath should be stirred continually to insure an even distribution of temper~ture throughout
the sample of oil.
2. When the -thermometer in the oil cup registers exactly 37.8C., remove
the cork from the outlet tube, starting the stop watch at the same instant,
and collect the effluent oil in the flask graduated to hold 60 cc. When the
level of the oil in the flask reaches the 60 cc. graduation mark, stop the
watch.
The time in seconds required for the efflux of 60 cc. of the liquid
petrolatum is called the Saybolt universal viscosity.
The U.S.P. requires that liquid petrolatum have a kinematic
viscosity of not more than 0.370 at 37.8C. (lOOF.). This value
is equivalent to a Saybolt universal viscosity of 170.
1. Consult the viscosity conversion table (U.S.P., page 477) and express
the results obtained in the above exercise in terms of kinematic viscosity
and in terms of Engler degrees.
2. If the viscosity of a sample of liquid petrolatum is found to be less than
thc official requirement, what does the determination indicate relative to
the composition of the oil?
3. Would viscosity requirements be of value as a constant for (1) glycerin,
(2) castor oil, (3) simple syrup?
CHAPTER XVII
PHOTOMETRIC METHODS OF ANALYSIS
Photometric methods of chemical analysis are based upon the

measurement of the changes in the amount or character of light
caused by chemical reactions. Changes in the amount of light
may be due to absorption or reflection. Measurements based
on the absorption of light are termed colorimetric methods, and
the process is known as colorimetry. Measurements based on
the reflection of light are termed nephelometric methods, and the
process is termed nephelometry.
FIG.
37.-Nessler apparatus.
Colorimetry.-The use of color as a means of determining the

amount of a given constituent in a substance has been utilized
for many years in analytical work. It is only in recent times,
l;lOwever, that .the methods have been extended and refined.
Colorimetric methods of analysis generally consist in adding
a reagent to a solution of the substance being tested so that a
color is produced and comparing the intensity of color in the
product with that produced by adding the same reagent to a
solution of the substance of known concentration. Frequently,
they are rapid and accurate; however, their greatest advantage
is their extreme sensitivity which makes it possible to determine
very small amounts of substance, in many cases unattainable by
other methods.
260
261
The simplest type of apparatus for colorimetric measurements

consists of the Nessler tubes and rack (Fig. 37). The tubes have
parallel sides and flat bottoms and are graduated to 50 and 100
cc. Nessler tubes should be of the same diameter, but frequently
they are not. Consequently, they should be compared for
equality of distance between the 50 and 100 cc. graduation marks
and only those which agree should be selected. The use of this
apparatus is illustrated in the following exercise.
Exercise 81
Object.-To Determine the Ammonia Content of Water.

Materials Required.-Natural water (use tap water).
Ammonium chloride solution. Dissoive 0.3141 Gm. of pure ammonium
chloride in ammonia-free water and dilute to 100 cc. Dilute 10 cc. of this
solution to 1,000 cc. with ammonia-free water. Each cubic centimeter of
the resulting solution contains the equivalent of 0.01 mg. of NH g
Rochelle salt solution. Dissolve 5 Gm. of pota~sium and sodium tartrate
in 10 cc. of water.
Nessler!s reagent. Dissolve 2.5 Gm. of potassium iodide in 3 cc. of
water, add 3.5 Gm. of mercuric chloride, and stir until solution is complete.
Add 100 cc. of a 25 per cent solution of potassium hydroxide, mix, allow to
settle, and decant the clear supernatant liquid. Preserve the solution in a
dark place.
Ammonia-free water. Test the distilled water in the laboratory by
placing 50 cc. in a Nessler tube previously washed several times with the
distilled water and add 2 cc. of Nessler's reagent. Cover the tube and
allow it to stand for 5 min. If the color produced after standing is more
intense than that of the ~essler's reagent immediately after dilution, the
water must be purified. If purification is necessary, add 1 Gm. of
potassium permanganate to 1,000 cc. of the distilled water; distil and when
the distillate does not give a test for ammonia with Nessler's reagent, collect
the water in a clean bottle previously rinsed with a portion of the ammoniafree distillate.
12 Nessler tubes.
A Nessler tube stand.
Procedure.-Place 50 cc. of the water to be tested in one Nessler tube.
Place 0.1, 0.2, 0.3, 0.4, 0.5, etc., cc. of the ammonium chloride solution in the
other eleven Nessler tubes and add ammonia-free distilled water up to the
50 cc. mark. Add 2 cc. of Rochelle salt solution and 2 cc. of Nessler's
reagent to each of the tubes. Mix the contents of the tubes thoroughly and
place them in the stand. While looking vertically down through the solution, vary the standard color solutions until the color of the unknown
matches one or lies between two of them. Calculate the milligrams per
liter of ammonia in the sample of water tested.
262
When comparing the intensity of color in the tubes, place them

in the stand and adjust the position of the latter in front of
a window so that light from the opaque reflector will pass upward
through the tubes.
Calcium and magnesium are precipitated from aqueous solution by Nessler's reagent. The Rochelle salt is added to prevent
this.
Nessler's solution reacts with the ammonia and ammonium
salts to form an orange-colored compound of the composition
HgO.Hg(NH2)I. When Nessler's reagent is added to solutions
containing more than a few
milligrams of ammonia per 100
cc., a distinct orange-colored
precipitate is formed, and very
small concentrations of ammonia, such as 1 part in 1,000,000,
produce a distinct yellow to
brown coloration.
The number of milligrams
of ammonia per liter of water
may be calculated as follows:
Assume that the color produced
in the sample exactly matches
that in the tube containing 0.5
cc. of the ammonium chloride
solution. Since each cubic
centimeter of ammonium chloride solution contains the equivFIG. 38.-Duboscq colorimeter.
alent of 0.01 mg. of NH3 per
cubic centimeter, 0.5 cc. will
contain the equivalent of 0.005 mg. per cubic centimeter or 5.0
mg. per liter. The sample of water must contain the same
amount or 5 mg. per 1,000 cc. equivalent to 5 parts of ammonia
in 1,000,000 parts of water.
1. What is meant by the term colorimetry?
2. Look up "Aqua" in the U.S.P. and explain the use of Nessler's reagent
in the tests for purity.
263
3. Look up Beer's law in a textbook of physics and explain what application if any is made of that law in colorimetric analysis.
4. Review the Gunning-Kjeldahl method of estimating nitrogen in organic
compounds and briefly explain how that method combined with the colorimetric method might be used to determine the nitrogen content in very
small amounts of certain organic compounds.
Colorimeters.-A number of colorimeters have been developed

for rapid and precise colorimetric measurements. The Dubo8cq
colorimeter (Figs. 38 and 39) is a
device of :this type, and since it is
generally used, its construction and
use will be described here. The
essential parts of the instrument
consist of plungers, cups, a reflecting
m i r r 0 r, a compound prism, an
observation microscope, and adjustments. Two prisms of optical glass,
hexagonal in form, matched for color
and with optically plane and parallel
ends are attached to the frame of
the instrument by threaded metal
adapters. Two detachable cups in
which the liquids under examination
are placed are threaded on movable
stages beneath the plungers. These
cups are made of glass fitted into I=~~~~
metal casings and have bottoms ~
made of optically inactive glass
disks. An adjustable mirror with
two reflecting surfaces is so placed
that light can be reflected through
the cups. A compound prism is
built in above the plungers in a
FIG. 39.-Diagram of the Dudust-proof housing in such a manner
boscq colorimeter showing the
as to bring the light beams through path
of light.
each cup to a common axis, light
from one cup illuminating one-half of a circular field and light
from the other cup illuminating the other half. An observation
microscope, by which the observer views both fields of illumination with one eye, is focused on the two fields. Adjustments
264
are provided whereby the cups may be raised or lowered, the

level of each cup being shown on a scale and vernier.
Exercise 82
Object.-To Determine the Amount of Epinephrine Hydrochloride in Solution of Epinephrine Hydrochloride.

Materials Required.--o.050 Gm. of pure epinephrine.
10 cc. of 1 per cent solution of potassium iodate.

Commercial epinephrine hydrochloride solution 1 to 1,000.
A Duboscq colorimeter.
Procedure.-l. Place 0.050 Gm. of pure epinephrine in a 50 cc. volumetric
flask, add 0.5 cc. of 0.1 N HCI, and dilute the solution with sufficient distilled water to make 50 cc.
This standard solution of epinephrine hydrochloride containing

1 part of epinephrine to 1,000 parts of solution must be prepared
freshly as needed.
2. Place 20 cc. of distilled water in a 50 cc. Erlenmeyer flask, add 5 cc.
of 1 per cent potassium iodate solution and 0.25 cc. of 0.1 N hydrochloric
acid, and warm the solution to 38C. Then add exactly 0.5 cc. of the
standard epinephrine solution and heat the resulting solution at about
38C. for exactly 15 min. Prepare another acid solution of potassium iodate
in the same manner, warm it to 38C., add exactly 0.5 cc. Vf the sample of
epincphrinc hydrochloride to be tested, and heat this solutlOn at 38C. for
exactly 15 min.
The greatest intensity of color is produced after the epinephrine

has been heated for about 10 min. at 38C. It is important
that the unknown and the standard solution be heated at the
same temperature for the same 'length of time.
3. Adjust the colorimeter, place it before an ample source of light, preferably diffused daylight, and regulate the mirror so that the two halves
of the field are of the same color and equal brightness. Fill one of the cups
about one-half full with the known solution and the other cup about onehalf full of the unknown. Replace the cups in the instrument and raise
them until the plungers come in contact with the bases. 4djust the liquid
depth in the cup containing the known solution to any convenient level,
preferably to an exact number of millimeters. Raise or lower the cup
containing the unknown liquid until both halves of the field are of the same
color and intensity. Take three independent readings of t)1e instrument
with the cups set at different levels. Calculate the percentage of epinephrine
in the unknown solution.
265
In adjusting the colorimeter, make certain that the plungers

and cups are fitted tightly. Raise the cups until the plungers
come in contact with the cup bases. The verniers should then
read zero. The best source of illumination is diffused daylight
obtained by placing the colorimeter before a north window,
but that prod1J.ced by an electric colorimeter lamp may be used.
The colorimeter cups should never be filled much more than
one-half full, otherwise their liquid contents may overflow when
the cups are raised. After placing the known and unknown
liquids in the colorimeter cups, raise the latter until the plungers
come in contact with the cup bases to force any bubbles of air
out from beneath the ends of the plungers.
In taking readings, set the level of the cup containing the known
liquid at some definite position, e.g., 15 mm., and regulate :,he
level of the cup containing the unknown liquid until a color
match is secured. Repeat this procedure with the cup containing
the known liquid set at 20 and 25 mm. readings. When the two
halves of the field of the colorimeter are of the same color and
intensity, the color intensities of the two solutions are inversely
proportional to the concentration. If C1 and D1 are the respective concentration and depth of the known solution, and C 2 and
D2 are the corresponding symbols for the unknown, then C 1 X
D1 = C2 X D 2 If C1 = 1,000 mg. per liter, D1 = 20 mm., and
D2 = 25 mm., then: 1,000 X 20 = 25 X C2, and
C2
= 1,00025X 20 = 800 mg. per l't

1 er,
the concentration of the unknown.

1. Is the principle involved in colorimetric measurements with the
Duboscq colorimeter the same as when Nessler tubes are used?
2. What advantages and disadvantages does the colorimetric method of
analysis of epinephrine have in comparison with the biological method?
3. Consult the reference at th(l end of this chapter, and give several
applications of colorimetric methods of analysis to pharmaceutical products.
Exercise 83
Object.-Assay of Crocus for Color.

Materials Requlred.-{).l Gm. crocus in fine powder.
266
10 cc. of 0.01 N potassium dichromate.

A Nessler apparatus or a colorimeter.
Procedure.-"Weigh accurately 0.1 Gm. of Crocus, in moderately fine
powder, and macerate it at room temperature in 100 cc. of distilled water for
three hours with frequent shaking. Immediately filter thc mixture and add
sufficient distilled water through the filter to make 100 cc. Dilute 10 cc. of
this filtrate, accurately measured, to 100 cc. with distilled water. Immediately compare the color of this solution, in Nessler tubes or in a colorimeter,
with tho color of hundredth-normal potassium dichromate.
"The color tint of the dilute Crocus solution approximates that of the
hundredth-normal potassium dichromate; the strength of color, w.hen
expressed in mm. of column height, shall be not less than that of the hundredth-normal potassium dichromate."
See Exercise 81 (page 261) for directions and explanation of the

use of the Nessler apparatus and Exercise 82 (page 264) for
directions and explanations of tl;te use of the colorimeter.
Cudbcar and tincture of cudbear of the N.F. arc also assayed
for color by similar procedures. Other simple color comparisons
are used in the official tests for carbonizable substances. These
qualitative tests are based upon matching the color developed
when a substance is treated with sulfuric acid with that of various
standard matching fluids. (See U.S.P. XI, page 95'7.)
1. What are colorimetric solutions? (See U.S.P.)
2. Look up the official tests for limit of lead and carbonizable impurities in
the U.S.P. Are these colorimetric tests?
Nephelometry.-Nephelometric measurements afford a means

of determining directly the mass or weight of a precipitate in
suspension, i.e., without filtering, drying, and weighing. The
method commonly used is based upon the measurement of the
brightness of the light reflected by a cloud of finely divided
particles suspended in a liquid. When other conditions remain
constant, the intensity of the reflected light is a function of the
amount of suspended particles.
.
Nephelometric methods of analysis have been applied to a
great number of inorganic and organic compounds, such as
chlorides, sulphates, ammonia, phosphorus, silver, proteins, fats,
volatile oils, and alkaloids. Many substances can be determined quantitatively by. nephelometric methods in concentrations
of 0.1 to 1 part or less in 1,000,000 parts of water; e.g., 1 part of
267
phosphorus in 330,000,000 parts of water has been determined

by this method. Gen~rally nephelometric methods are limited
to measurements of suspensions of substances in low concentration, usually not greater than 100 mg. per liter. Larger amounts
of substances can be estimated, however, if properly diluted.
The instrument used for nephelometric measurements may
consist of a Duboscq colorimeter fitted with special nephelometer attachments. These attachments include two cups with
transparent sides and black bottoms in place of the colorimeter
cups; two tubular metal light shields which surround the cups in
such a way that the depth of the liquids illuminated by a light
source may be varied; a special nephelometer illuminator; two
condensers by means of which a pair of parallel rays are projected
horizontally into the cups containing the liquids under examination. Nephelometric determinations preferably should be carried
out in a dark room.
Exercise 84
Object.-To Detenpine the Amount of Arsenic Trioxide in

Solution of Arsenous Acid.
Materials Required.-25 cc. of potassium molybdate solution, 1 per cent.
50 cc. of 1.0 N hydrochloric acid.
25 cc. of cocaine hydrochloride solution, 2 per cent.
0.1 Gm. of pure arscnous acid.
10 cc. of solution of arsenous acid.
A Duboscq nephelometer with illuminator attachments.
Procedure.-1. Place 25 cc. of 1 per cent potassium molybdate solution
in a 100 cc. flask. add 50 cc. of 1.0 N hydrochloric acid and 25 cc. of 2 per
cent cocaine hydrochloride solution. Shake the mixture thoroughly and
preserve it in a tightly stoppered bottle in a dark place.
This mixture constitutes the nephelometric reagent for arsenic.

It will keep for a long time if properly preserved. Immediately
prior to using a portion of this sc;>lution, it should be filtered from
any sediment present, using a quantitative filter paper.
2. Dissolve exactly 0.1 Gm. of pure arsenic trioxide, previously dried to
constant weight at 1000., in sufficient distilled water to make 100 cc.
This is a standard arsenic trioxide stock solution containing

1 mg. of As 20 a in each cubic centimeter. Other standard
268
solutions containing 0.1,0.01,0.001 mg. of As 20 S can be prepared

by dilution in the proper ratio.
3. Dilute 10 cc. of the standard arsenic trioxide solution to 1,000 cc. with
distilled water. Place 10 cc. of this diluted solution in a 30 cc. glassstoppered bottle, add 2.5 cc. of the cocaine-molybdenum reagent, and shake
the mixture vigorously. Dilute 1.0 cc. of the unknown arsenous acid solution to 1,000 cc. and treat a 10 cc. portion of this solution with the cocainemolybdenum reagent in the same manner and at the same time as the
standard.
The diluted standard arsenic trioxide solution should contain

0.001 mg. of As 2 0 a per cubic centimeter and the amount of
arsenic trioxide in the solution of arsenous acid should contain
approximately the same amount of As 2 0 a per cubic centimeter
if the solution is of about the official strength.
The cocaine-molybdenum reagent produces a cloud when added
to solutions containing arsenic trioxide. The best clouds are
obtained when from 0.05 to 0.005 mg. of arsenic are present in
each 10 cc. of the solution.
4. Pour the known and unknown solutions of arsenic trioxide into the
respective cups, filling them to about 6 mm. from the tap. Place the
tubular light shields about the cups on the cup stage and raise them up
until the scale reading is zero. Place the instrument in position wit~ respect
to the nephelometer illuminator, set the position of the cup containing the
known solution at 15 mm., and adjust the level of the cup containing the
unknown until a balanced field is obtained. Take three separate measurements with the cup containing the standard solution set at 15, 20, and
25mm.
The calculations are based on the principle that the turbidities

are inversely proportional to the length of the column of liquid.
Thus, if 0 1 is the concentration of the standard solution, O2
the concentration of the unknown, L1 and L2 the lengths of the
columns of liquids in the cups containing the known and unknown
solutions, respectively, the concentration of the unknown solu.
tion may be found by substituting in the following formula:
O2 = 0 1 X L1
L2
1. Would it be practicable to replace the present official test for arsenic
(see U.S.P., page 436) by a nephelometric method?
269
2. Name several official, arsenic-containing preparations to which this

method with slight modifications might be applied.
Exercise 85
Object.-To Determine the Amount of Oil of Peppermint in

Spirit of Peppermint.
Materials Required:-5 cc. of spirit of peppermint.
1 cc. of oil of peppermint.
Diluted hydrochloric acid.
Alcohol, 95 per cent (redistilled).
Procedure.-Dissolve 1 cc. of oil of peppermint, accurately measured
from a pipette, in sufficient 95 per cent alcohol to make exactly 10 cc.
Dilute 5 cc. of this solution with 25 cc. of diluted hydrochloric acid, shake
thoroughly, and compare by means of a nephelometer the turbidity produced with that of an equal volume of spirit of peppermint diluted in the
same manner.
When the alcoholic solution of volatile oil is diluted with

. hydrochloric acid, the volatile oil is thrown out of solution in
the form of finely divided globules with the formation of a fairly
stable emulsion. The amount of oil present is then estimated by
measuring the amount of light reflected from a column of the
emulsion by matching the latter with the light reflected from a
column of emulsion containing a known amount of oil. The
measurements should be made as soon as possible after throwing
the oil out of solution so that there will be no error caused by
breaking of the emulsion.
Turbidimetric Tests.-Turbidimetric tests are applied to
certain official chemicals to insure the absence of excessive
amounts of chloride and sulfate. The following directions are
given in the U.S.P. for the performance of these tests:
"In carrying out these turbidimetric tests, the following points
are to be observed: The same quantities of the sam'e reagents
must be used in the test of the substance under examination and
in the control test. The glass cylinders in which the tests
are made must be of the same diameter and match in all other
respects as closely as possible. The precipitating reagent must
be added to both in immediate sequence.
"Chloride.-The prescribed quantity of the substance to be
tested is dissolved in from 30 to 40 cc. of distilled water, and the
solution neutralized, if necessary, with nitric acid, using litmus
270
paper as the indicator. One cubic centimeter of nitric acid and

1 co. of silver nitrate T .S. are added and then sufficient distilled
water to make 50 cc. After mixing well and allowing to stand
five minutes protected from direct sunlight, the turbidity, if
any, is compared with that produced'in a control test made with
the specified volume of fiftieth-normal hydrochloric acid.
II Bismuth Salts are first dissolved in a few cubic centimeters of
distilled water and 2 cc. of nitric acid, then diluted with distilled
water to 50 ce.
II Sulfate.-The specified quantity of the substance to be tested
is dissolved in from 30 to 40 cc. of distilled water, and the solution
neutralized, if necessary, with hydrochloric acid, using litmus
paper as the indicator. One cubic centimeter of diluted hydrochloric acid and 1 cc. of barium chloride T.S. are added and then
sufficient distilled water to make 50 cc. After mixing well,
it is allowed to stand for ten minutes and the turbidity, if any,
is compared with that produced in a control test made with the
specified volume of fiftieth-normal sulfuric acid.
"If the solution, after the addition of acid, its not perfectly
clear, it is filtered through a filter paper free from chloride or
sulfate, then the silver nitrate or the barium chloride is added.
II In applying the turbidimetric tests to salts of heavy mrta1s,
which normally show an acid reaction, their aqueous solutions
prepared for the test are not to be neutralized.
"When the tests for chloride or sulfate are to be applied to a
specified volume of a solution of a substance prepared as directed
in the text, and the permissible limit for these impurities corresponds to 0.2 cc. or less of fiftieth-normal hydrochloric or
sulfuric acid, the solution is not to be further diluted. The control test is also made with the same volume of water (or other
specified solvent)_as in the test."
Exercise 86
Object.-Limit Test for Chloride and Sulfate in Calcium

Gluconate.
Materials Required.-3 Gm. of calcium gluconate.
Silver nitrate T.S. (0.1 N silver nitrate).
271
Barium chloride T.S. (12 Gm. of barium chloride in sufficient water to

make 100 ce.)
Nessler apparatus.
Procedure.-Proceed as directed above using 1 Gm. of calcium gluconate
and 1 cc. of 0.02 N HCI in the test for chloride and 2 Gm. of calcium gluconate and 1 cc. of 0.02 N H 2S0 4 in the test for sulfate.
1. Calculate the per (lent of chloride and of sulfate permitted in calcium
gluconate in the official test for purity.
2. List several official substances which are required to show no turbidity
when tested for chlorides and sulfates.
~. How could the volatile oil content of a highly colored solution be
determined nephelometrically?
4. The presence of sugar in a solution containing volatile oils alters the
particle size when the oil is thrown out of solution. How could the volatile
oil content of a cordial be determined by means of the nephelometric method?
5. Acetone is precipitated from solutions containing as little as 1 part in
100,000,000 by silver-mercury cyanide solution. Could the nephelometric
method be used to estimate the presence of small amounts of acetone in
alcohol, in ether, and in urine?
6. How might minute quantities of each of the following ions be determined: iodide, sulfate, phosphate, and calcium?
7. Could minute quantities of alkaloids in solution be estimated nephelometrically? Suggest possible precipitants.
Those who desire to make a more extended study of photometric, methods of analysis will find a very comprehensive treatment
of the subject and an extensive bibliography in "Photometric
Chemical Analysis," by J. H. Yoe (2 vols., John Wiley & Sons,
Inc., New York, 1929).
CHAPTER XVIII
Electrometric methods of analysis may be divided into two

classes, namely: (1) conductimetric methods in which the end
point is detected by measuring the change in electrical conductance during a titration; (2) potentiometric methods in which
the end point is found by measuring the change in potential of a
suitable electrode during the titration. The latter has become
of increasingly great importance in all fields of chemistry within
comparatively recent years.
Portions of the theoretical considerations of pH which
follow have been taken with slight changes from the U.S.
Pharmacopoeia.
Many of the compounds which are pharmaceuticaLly important
are acids, bases, or salts thereof. In aqueous or hydro alcoholic
solution they tend, in great or small degree, to dissociftte into
their respective ions. The concentration and activity 01 hydrogen ions in a solution influence the concentrations of the anions,
cations, and undissociated molecules present in the solution.
These factors, in many instances, affect the stability, therapeutic
activity, and pharmaceutical elegance of medicaments in aqueous
or hydroalcoholic solutions.
Acid-Base Equilibrium and pH.-The dissociation of acids
may be represented by the type description:
AH
H+
+ A-
In a similar manner the dissociation of substituted ammoniums

may be represented:
BHI
IB +H+
The dissociation of the metallic hydroxides may be represented:

BOH
B+
272
+ OH-
273
For the purpose of this discussion, salts, certain acids, and the
hydroxides of the alkali metals are assumed to be completely
dissociated in dilute aqueous sE>lution.
At equilibrium the ordinary mass action equation will hold as a
first approximation for weak acids
[H+][A-] = Ka
[HA]
[A-]
Ka
[HA] = [H+]'
(1)
(2)
where the brackets indicate molar concentration of each ionic or

molecular species.
The constant, K a , is the dissociation constant of the acid. The
value of Ka is' markedly different for various acids, and for the
same acid it is appreciably affected by the presence and concentrations of various ions in solution. The ratio [A-]/[HA] in
equation (2) is seen to depend upon the value of Ka and the concentration of hydrogen ions. Assuming that Ka remains constant, the ratio [A-]/[HA] can be changed only by changing the
value of [H+]. This can be accomplished by the addition of a
strongly dissociated acid which increases [H+] or the addition of a
strongly dissociated base which will decrease the value of [H+].
Furthermore"the ratio [A-]/[HA] may be changed by the addition of the acid itself or its salt, but not directly as the ratio of
these added quantities, for [H+] will have been changed also.
The magnitudes of [H+] vary enormously, i.e., from one to one
ten-inU!ionth squared or 10- 14 The desirability of an exponential or logarithmic scale of expression is at once apparent. The
system generally adopted is that devised by S~renson, namely,
the use of the symbol pH, which is defined as the negative logarithm
of the hydrogen ion concentration (see page 67). Hence:
pH = -log [H+]
or
log [H+]
(3)
The relation of pH to [H+] may be seen in Table XXXVII.

The logarithmic nature of this system should be borne in mind
constantly in considering changes in hydrogen ion concentration
and its connotation in terms of pH as illustrated in Table
XXXVIII.
274

TABLE XXXVII
pH
Normality,
hydrogen ions
Normality,
hydroxyl ions
1
10-1
10-2
10-3
10-<
10-'
10-
10- 7
10-s
10-'
10-1
10-11
10- 12
10- 13
10-14
10-14
10-13
0
1
2
3
4
5
6
Neutral potnt 7
8
9
10
11
12
13
14
10-12
10- 11
10- 10
10-
10-8
10-7
10-
10-'
10-4
10-3
10- 2
10- 1
1
TABLE XXXVIII
[H+]
pH
CorrespondIng Change
COO
0.70
O.~O
1.5
1.1
1.05
1.023
0.04
0.02
0.01
Times Increased or Decreased

10
0.18
Equation (2) may be written

[A-]
log [HA]
+ log Ka
(4)
= pH
Obviously when the ratio [A-]/[HA] is unity, log

equal to pH.
Also, when pH is greater than log
ia
ia
will be
by two units,
practIcally the entire compound exists as species A-.

however, is less than log
ia
When pH,
by two units, practically the entire
compound exists as species HA.
When pH has a value lying
within
2 units log
i,,'
275
there will be relatively large proportions
of each species present\

The standard device generally used in the determination of
hydrogen ion concentration is the hydrogen half-cell.
A hydrogen half-cell consists of a noble metal electrode,
immersed in a solution containing hydrogen ions or substances
capable of supplying such ions at the activity symbolized by
(H+), under a definite partial pressure of hydrogen, PH2, and supplied with a catalyst such as platinum black to facilitate the
half-reaction
H2
2H+
+ 2e
When two such half-cells are placed in liquid junction, the

electromotive force E of the cell is defined by
E = RT In (H+)'yPH2
F
(H+)yP'H2
+E
(5)
L,
Where (H+)' is the activity of the hydrogen ions in the halfcell having the greater hydrogen ion activity, P'H2 is the hydrogen
pressure in the same half-cell and EL is the potential jump at the
liquid junction of the half-cells. The latter is not eliminated
but is reduced to a small, usually neglected value, by making the
junc,tion with saturated potassium chloride solution.
The ultimate standard of reference is the hydrogen half-cell
in which the hydrogen ion activity is :unity and the hydrogen
pressure is one atmosphere. If this half-cell were used in conjunction with another half-cell of lower hydrogen ion activity and
the hydrogen pressure were the same in the two half-cells, then
RT
1
E = FIn (H+)
or
~
1
E = 0.0581 log (H+)
(6)
(7)
The foregoing ultimate standard of reference is not used in

practice and it is impossible either to eliminate entirely the liquid
junction potential or to calculate it with exactitude. Therefore
276
for official purposes the following is adopted for approximate

measurements.
Hg \ Hg 2Cb, KCI (sat. soln.)
IunSkolution ofH
IPt,
p
~nown
H2 (1 atmos.)
Employing this cell at 20C., pH may be calculated by the

following equation:
pH
E - 0.2488
0.0581
(8)
The electromotive force is measured by meane of a potentiometer.

When the standard 0.05 M potassium biphthalate solution
(page 296) is employed in the foregoing cell, in place of the solution of unknown pH at 20C., the cell has an electromotive force
of 0.4797 volt, corresponding to pH 3.974.
These half-cells employed under the prescribed conditions constitute the reference device for pH measurements for the official
solutions. For official purposes, various electrodes may be
employed at the discretion of the operator which are capable of
as great a degree, or a greater degree, of acg.uracy than that
set forth in the colorimetric method, namely, 0.1 pH.
When the electrometric method is employed for hyqroalcoholic
solutions and the electromotive force developed converted into
pH, the conversion is based on the assumption that the equation
holds for hydroalcoholic solutions as it does for aqueous solutions.
Strictly speaking this condition does not obtain. Therefore
the values expressed as pH are relative and subject to change with
varying concentrations of alcohol.
The term pH is expressed generally by an integer and two
significant decimals. The reliability of the value of the figures
in the second decimal place is dependent largely on the method
employed in carrying out the determination. For official purposes, the expression is given with one decimal only. For practically all purposes, this degree of accuracy is considered sufficient.
However, the method described is capable of a higher degree of
accuracy.
The potentiometric method of analysis may be applied to
determine the end point of titrations and to determine the
hydrogen ion concentration of solutions. The hydrogen ion
277
concentration of solutions can also be determined colorimetrically (see page 297). The potentiometric method, however, is
more accurate in general and is to be preferred even though it
requires a more elaborate apparatus and more experience on the
part of the analyst.
Potentiometric Methods.-When an acid or an alkali is dissolved in water, the characteristic properties of the solution
obtained are due to the hydrogen and hydroxyl ions formed by
the dissociation of the acid or base. The effective acidity
or alkalinity depends on the extent of dissociation of the acid
or base in solution and is independent of the total acidity or
alkalinity. In the volumetric estimations of acids and alkalies,
the total acidity or alkalinity is determined. In measuring
the effective acidity or alkalinity (the strength of the acid or
base) of a solution, however, it is necessary to determine the
hydrogen ion concentration.
The hydrogen ion concentration of an aqueous solution may be
determined by measuring the voltage developed between two
electrodes of special character immersed in the solution by means
of a potentiometer, since the voltage developed between the
electrodes directly depends on the concentration of hydrogen
ions in thc solution. The measurement of the voltage produced
between the electrodes is equivalent, therefore, to a determination
of the hydrogen ion concentration and to the determination of the
effective acidity or alkalinity of th~ solution.
The electrodes employed in the determination of hydrogen ion
concentration must be of such character that one of them will
develop a potential that varies according to the concentration of
hydrogen ions, and the other must have a constant potential
unaffected by the hydrogen, ion concentration. Although a number of electrodes of different character have been developed for
use in potentiometric methods of hydrogen ion concentration
measurements, the fundamental method involves the use of the
so-called hydrogen electrode an'd. the saturated calomel electrode
described here.
The hydrogen electrode (Fig. 40) consists of a small square of
platinum foil attached to a short piece of platinum wire sealed
into and extending through one end of a glass tube. Mercury
is poured into the open end of this tube to cover the platinum wire
278
in the bottom so that the electrode may be connected with

the potentiometer by dipping a wire in the mercury. This
electrode element is placed in another tube of larger diameter
through a side tube of which purified hydrogen gas is conducted
to the electrode when it is immersed in a solution.
Holes in the side of this outer tube at the level of
the electrode element permit the hydrogen gas to
escape without forcing the solution completely out
of contact with the platinum.
When the platinum tip of this electrode is coated
with platinum black (platinum in a finely divided
state), the platinum black absorbs and holds a
relatively large volume of the gas. Consequently,
when an' electrode prepared in this manner is
saturated with hydrogen gas and immersed in a
FIG. 40.- solution containing hydrogen ions, it behaves as a
~e~t~~;e~ e n hydrogen electrode and develops a potential that
is definitely related to the hydrogen ion concentration of the solution.
To platinize the hydrogen electrode, clean a new electrode by
dipping it in hot solution of potassium dichromate, then in
concentrated sulfuric acid, and finally wash it thoropghly with
distilled water. This procedure removes any organic matter
adhering to the platinum. The platinum black can be removed
from old electrodes by electrolysis in concentrated hydrochloric
acid, the electrode being used as the anode. In the latter case,
the electrode should be removed from the hydrochloric acid and
washed with water as soon as all of the platinum black has
dissolved; otherwise the platinum of the electrode will go into
solution. Dissolve 3 Gm. of platinic .chloride and 0.010 Gm. of
lead acetate in sufficient distilled water to make 100 cc. Immerse
the electrode in this solution and connect it as the cathode to two
dry cells connected in series (the positive, center pole, of one cell
connected to the negative, outside pole, of the other) and use a
clean platinum wire as the anode to complete the circuit in the
solution. Allow the current to pass for from 3 to 5 min. or until a
thin, even deposit of platinum black covers all of the electrode
so that the sheen of metallic platinum cannot be observed.
When platinization is completed, wash the electrode several times
279
with distilled water, immerse it in dilute sulfuric acid, and electrolyze as cathode for 1 or 2 min. to saturate the platinum black
with hy9-rogen. Suspend the electrode in distilled water until it
is used.
The calomel electrode (Fig. 41) is prepared as follows: Add 200
Gm. of pure potassium chloride to 500 cc. of hot distilled water
contained in a 1,000 cc. beaker, add 0.5 Gm. of pure mercurous
chloride (calomel) to the hot solution, and stir the mixture well.
Allow the mixture to cool to room temperature and filter out the
e~cess potassium chloride and calomel. Store the solution in a
well-stoppered bottle for future use. Clean the electrode
vessel thoroughly by washing it successively with cleaning
mixture and distilled water. Dry the vessel. Pour pure mercury into the electrode vessel until the platinum wire in the
bottom is covered. Add a layer of mercurous chloride about a quarter ineh deep
over the mercury, pour some of the potassium chloride-calomel solution 0 V e r it, +
stopper the mouth of the vessel, close the
stopcock, and shake the contents together,
being careful to avoid as far as possible
getting mercury and calomel into the side
arm. Allow the mixture to settle and fill
the vessel and side arm with the potassium
chloride-calomel solution, wash away any
floating calomel by overflowing, make ~_flL"DINARY M<IOCU".
certain that no air bubbles are trapped FIG. 41.-Calomel eleotrade.
in the side arm, and close the stopcock.
Stopper the electrode vessel until it is ready for use. Fill
the side tube connecting with the bottom of the vessel with
ordinary mercury so that the platinum wire that extends into
this tube from the vessel is covered.
Only the purest obtainable chemicals should be used to prepare the calomel electrode. Specially purified mercury, calomel
produced by electrolysis, and analyzed potassium chloride can
be obtained from supply houses for this purpose.
The potential developed by the calomel electrode depends on
the concentration of potassium chloride used and is independent
of the hydrogen ion concentration of the solutions examined
280
during a determination. The potential also varies with the

temperature of the electrode. The following table shows the
potentials developed by the three commonly used calomel
electrodes at different working temperatures:
TABLE
XXXIX
Potential at
Calomel eleotrode
20C.
0.1 N KCl. ...................
NKCl. ......................
Saturated KCl. ...............
0.3379
0.2860
0.2496
25C.
0.3376
0.2848
0.2458
30C.
0.3371
0.2835
0.2420
The scheme of assembly for hydrogen ion measurements is

shown in Fig. 42. A is a beaker of 50 to 200 cc. capacity containing the solution being investigated. B is the hydrogen
FIG. 42.-Scheme of assembly for hydrogen ion measurements.
electrode. C is a cylinder of compressed, electrolytic hydrogen

equipped with a pressure-reducing valve D. The gas supply
is passed through a solution of pyrogallol in potassium hydroxide
contained in the gas-washing bottle E and then through distilled
281
water contained in the second gas-washing bottle F and finally

into the hydrogen electrode vessel. The hydrogen gas may be
generated from zinc and sulfuric acid, but in this case, the
hydrogen should be wash~d by passing it through successive
gas-washing bottles containing alkaline permanganate, alkaline
pyrogallol, sulfuric, acid, and water to remove arsenic, hydrogen
sulfide, etc. All of the connections in the gas train should be
tight to exclude the entrance of air. Pure gum rubber tubing,
5 mm. bore, and of medium wall should be used for the connections. H is an aspirator bottle containing a reserve supply of
saturated potassium chloride-calomel solution connected to the
calomel electrode vessel so that the side arm of the latter can be
flushed out at frequent intervals and before each new determination. The flow of solution from the aspirator bottle is
controlled by the pinchcock I.
The voltage measuring equipment consists of a potentiometer
by means of which the voltage produced by the hydrogen and
calomel electrodes dipping into the solution is measured by
establishing a 'known voltage in opposition and exactly equal to
that of the electrodes; a storage cell, or battery, K to supply a
constant source of current; a galvanometer J which shows when
the voltage developed at the electrodes is exactly counterbalanced against the voltage of the potentiometer; and a standard
cell L to establish known voltage conditions in the potentiometer
circuit.
Exercise 87
Object.-To Determine the End Point of Titration of Hydrochloric Acid with Sodium Hydroxide Potentiometrically.
Materials Required.-A potentiometer.
A galvanometer.
A standard cell.
A storage battery.
A hydrogen electrode.
A calomel electrode.
A cylinder of compressed electrolytic hydrogen 100 or 200 cu. ft. capacity,
equipped with a pressure-reducing valve.
Two gas-washing bottles.
An aspirator bottle.
12 to 20 ft. of well-insulated, IS-gage copper wire.
3 titration beakers, 50 to 200 cc. capacity.
282
3 ft. of rubber tubing, 5 mm. bore, medium wall.

8 oz. of specially purified mercury.
1 oz. of calomel, specially purified.
1 lb. of potassium chloride, analyzed grade.
3 Gm. of platinic chloride.
0.010 Gm. of lead acetate.
50 cc. of normal sodium hydroxide solution.
100 cc. of normal acetic acid solution.
25 cc. of 0.1 N hydrochloric acid.
0.1 N sodium hydroxide solution.
A number of types of potentiometers are available and used.

The potentiometer referred to here is the Leeds Northrup and
Company modified type K instrument. The student potentiometer made by the Leeds Northrup and Company is a simpler
and less expensive apparatus.
Procedure.-l. Assemble the apparatus, as illustrated in the diagram
(Fig. 42). Close the switch on the potentiometer that connects the standard
cell in place of the electrodes, tap the galvanometer key gently, and note
the deflection of the galvanometer. Adjust the rheostat R until the galvanometer index does not move when the key is tapped.
The voltage of the potentiometer is exactly balanced against

that of the standard cell, the voltage of which is known, when
the galvanometer index does not move upon tapping the galvanometer key. Do not hold the galvanometer key down to note
t}{e deflection of the index, but tap it firmly and quickly.
2. Dissolve 40.836 Gm. of reagent potassium biphthalate in distilled
water and dilute the solution to 1,000 cc. Dilute 25 cc. of this solution to
100 cc. to obtain 100 cc. of 0.05 M potassium biphthalate. Transfer about
25 cc. of this solution to the titration beaker and immerse the hydrogen and
calomel electrodes in the solution as indicated in the diagram (Fig. 42).
Disconnect the gas train from the hydrogen electrode, open the valve of
the pressure gage, and allow a current of hydrogen to flow for about 2 min. to
sweep all air out of the tubes and wash bottles. Connect the gas line to the
hydrogen electrode immersed in the buffer solution, and regulate the flow
of hydrogen so that about three bubbles of gas escape from the electrode per
second. Permit the hydrogen to flow for about 5 or 10 min. to saturate the
electrode and the solution with gaseous hydrogen. Flush a small quantity
of potassium chloride through the side arm of the calomel electrode, to
remove any test solution diffused into it, and close the stopcock on the side
arm almost completely. Change the switch on the potentiometer from the
standard cell to the electrodes, tap the galvanometer key, and note the
deflection of the index. Adjust the dial switch and slide-wire knobs on top
283
of the potentiometer plate until the galvanometer index shows no deflection

upon tapping the galvanometer key.
The voltage developed by the calomel and hydrogen electrodes

in this solution at 20C. is 0.4797 volt with a saturated calomel
electrode corresponding to pH = 3.974. If the apparatus
measures the required voltage in this solution, it is in proper
working order for measurements in solutions of unknown hydrogen ion concentration.
Do not change the rheostat adjustment while testing the
apparatus. Record the voltage from the setting of the dial
switch and slide wire.
3. Check the potentiometer voltage against the standard cell again as in
step 1, and if the galvanometer index is deflected, adjust the rheostat so
that the index does not deflect. Remove the test solution from the titration
beaker, wash the hydrogen electrode with distilled water, and flush a small
quantity of potassium chloride through the side arm of the calomel electrode.
Place 25 cc. of 0.1 N HCI in a 200 cc. titration beaker and dilute it with
about 80 cc. of distilled water. Determine the voltage developed by the
hydrogen and calomel electrodes immersed in the acid solution. Accurately
measure about 20 cc. of 0.1 N sodium hydroxide solution into the titration
beaker and again determine the voltage. Add successively four accurately
measured 1 cc. portions of 0.1 N sodium hydroxide and determine the voltage
after the addition of each cubic centimeter. Continue to add portions
of sodium hydroxide solution dropwise and determine the voltage after
the addition of each portion until 26 cc. have been added. Add 5 cc. more
of 0.1 N alkali and determine the voltage. Plot a curve using cubic centimeters of 0.1 N sodium hydroxide solution as abscissae and voltage readings
as ordinates, and determine the end point of titration from the curve.
Calculate the hydrogen ion concentration and the pH at the end point.
If the normality of both the acid and alkali solutions used is

not known, it is advisable to run a preliminary test, using a
suitable indicator to determine the end point so that the equivalent point determined potentiometrically will not be overrun.
As alkali is run into the acid in the titration, it will be noticed
that the voltage changes slowly at first, more or less gradually
as the equivalent point is approached, very rapidly with respect
to very small amounts of sodium hydroxide at the equivalent
point, and then very slowly again as an excess of alkali solution
is added.
The pH value of the solution corresponding to the measured
voltage at any point in the titration may be found from a curve
284
showing the relationship (Fig. 43). If the voltage is measured

with a hydrogen electrode and a calomel electrode at 250. the
pH value can be calculated from the following:
pH = 16.9 (V - 0.246) for the saturated calomel electrode
pH = 16.9 (V - 0.285) for the normal calomel electrode
pH = 16.9 (V - 0.338) for the 0.1 normal calomel !=Jlectrode
o ABC
'~ (
\ \\
Z
3
4
5
pH
6
7
8
9
10
1
12
13
\\1\
\~\
~ \\
Curve A ~Wifh Saturated

KCl Calomel Electrode
Curve 8-Wifh Normal KCl
Calomel Electrode
Curve C-WithTenthNormaJ
Calomel Electrode
\\\
'\ \\
1\\\
\ l\\
.~ l\
~\\
"\ \\
\\\
\
l\\
\) l\
140.2 0.3 0.4 0.5 0.9 0.1 0.8 0.9 1.0 1.1 1.2
Voltage
FIG. 43.-Graph showing relation of pH and voltage measured with hydrogen
and calomel electrodes.
V is the measured voltage, 0.246, 0.285, and 0.338 are the

reference potentials of the saturated, normal, and 0.1 N calomel
electrodes, respectively, and 16.9 is a factor which when multiplied by this difference converts the potential of the hydrogen
electrode which corre~ponds to the hydrogen ion concentration
of the solution into the corresponding pH value. The relationship between pH and voltage measured at 250. with the hydrogen and calomel electrode is given in the graph (Fig. 43). Some
typical titration curves are illustrated in Fig. 44.
285
Notes and Precautions.-l. A thin deposit of platinum black

on the hydrogen electrode is better than a thick deposit. The
former establishes equilib'rium more rapidly and is not "poisoned" so easily as the latter.
2. It is advisable to test different hydrogen electrodes against
the same calomel electrode in the same solu.tion for agreement in
3 4 567 6 9 W
~ n
E IT ffi
I./Oee. H2 S04 -O.1 Norm/

NaOH
II.
IOcc-He/-al Normal NH4 0H

1Occ.AceficAcid 0./ Nor-
IJ/.
ma/NaOff
IOcc.Orlhophosphoric
Acid-O.! NormaJ NaOIi
V. iOcc.NeQr/ySaturtried
BoricAcia-Norma/ NaOH
5~
6~
<
7~
9~
IO~
II;
IZ~
13:
14'
FIG. 44.-Charaotcristic titration curves.
Vertioal scale shows pH values.
voltage indications. A difference in the agreement between two

hydrogen electrodes can also be detected by substituting one
for the calomel electrode and mea'suring the other against it.
The agreement in the voltage of two calomel electrodes can also
be determined in this way by substituting one for the hydrogen
electrode.
3. The true potential of the hydrogen electrode in a solution
is attained only when the electrode and solution are saturated
286
with hydrogen. Inconsistent results are frequently' obtained in

hydrogen ion determinations due to the use of insufficient
hydrogen.
4. The hydrogen electrode cannot be used in solutions containing strong oxidizing agents, such as ferric iron, dichromates,
nitric acid, and chlorine. It cannot be used in solutions containing strong reducing agents, such as hydrogen sulfide or
sulfurous acid.
5. Certain organic compounds which are easily reduced
frequently prevent accurate results, e.g., unsaturated acids,
nitrobenzene, and aniline dyes.
6. The hydrogen electrode does not function properly in
solutions containing metal ions that fall below hydrogen in the
electromotive series of the metals or lead ions, since all of these
ions are reduced on the platinum electrode.
7. Sometimes unexpected and erratic results occur in determinations employing the hydrogen electrode. This may be due
to the so-called poisoning of the electrode which usually causes
a drift in the voltage readings when attempts are made to balance
the potentiometer. A number of substances are commonly
known to produce electrode poisoning, especially- mercury, proteins, oxygen, some alkaloids, formaldehyde, ammonia, and
I
hydrogen sulfide.
8. Hydrogen electrodes that have been poisoned can frequently
be restored to normal by washing them in nitric acid aI).d caustic
alkali solution successively and then electrolyzing them in sulfuric
acid solution.
1. Study one of the first three references given at the end of this chapter
and explain the principle of the potentiometer.
2. Explain why a platini.zed platinum electrode saturated with hydrogen
gas acts as a hydrogen electrode.
3. Consult the recent journal literature (during two preceding years)
and briefly summarize the articles treating 'of the application of hydrogen
ion concentration measurements to pharmaceutical products.
4. In the potentiometric titration of 25 cc. of 0.2680 N H 2S0 4 with NaOH
solution, using the hydrogen electrode and 'it saturated calomel electrode,
the following are values in millivolts for the corresponding volumes of N aOH:
0.0 cc. = 370.0; 5 cc. = 378.1; 10 cc. = 388.0; 15.0 cc. = 397.6; 20 co.
406.0; 25.0 ce. = 420.2; 28.0 cc. = 460.1; 30.0 cc. = 516.1; 30.5 cc. =
287
690.5; 31 cc. = 860.1; 35.0 cc. = 949.5; 40 cc. = 965.9; 45 cc. = 982.2;
50 cc. = 992.1. Plot the millivolts of electrode potential as ordinates and
the cubic centimeters of NaOH as abscissae. Determine the pH value at
the end point from the curve and calculate the normality of the sodium
hydroxiae solution.
Exercise 88
'Object.-To Determine the End Point of Titration of Acetic

Acid with Sodium Hydroxide Potentiometrically.
Materials Required.-The same as in Exercise 87, except that the 0.1
N hydrochloric acid is replaced by a solution of acetic acid of unknown
concentration.
Procedure.-Proceed as in Exercise 87.
1. Plot a curve using pH values as ordinates and cubic centimeters of

alkali as abscissae.
2. Look up the pH at which phenolphthalein and methyl orange change
color, mark the corresponding positions on the curve obtl\ined, and explain
which indicator is suitable for use in the titration of acetic acid with sodium
hydroxide.
3. Why does the voltage increase more rapidly at the beginning of the
titration in this case than it does in the titration of hydrochloric acid with
sodium hydroxide?
Exercise 89
Object.-To Determine the- pH of Saturated Boric Acid Solution, and Physiological Salt SolutIon.
Materials Required.-25 or 50 cc. of each of the above solutions.
The titration apparatus described in Exercise 87.
Procedure.-Determine the pH of each solution separately. Place the
respective solutions in the titration beaker, allow the hydrogen gas to flow
until the electrode has come to equilibrium with the voltage. Take three
voltage readings at 1 min. intervals. Calculate the pH of the solution from
the data obtained.
Since the concentration of the hydrogen gas in the electrode is

constant, the potential developed yaries with the concentration
of hydrogen ions in the solution and therefore may be used as a
measure of the concentration of the latter.
1. Name several official preparations to which the above method of
determining pH could be applied.
288
A considerable number of special electrodes have been

developed for use in potentiometric titrations in the place 'of the
hydrogen electrode. Thus a plain platinum wire used in place
of the hydrogen electrode is employed in oxidation-reduction
reactions. In pharmaceutical analysis, the quinhydrone electrode has been. used quite extensively in place of the hydrogen
electrode when difficulties are encountered in the use of the
latter.
The quinhydrone electrode consists of a piece of platinum wire
or foil, exactly the same as that used for the hydrogen electrode,
but the surface is not platinized or supplied with gaseous hydrogen, and it dips into a solution of quinhydrone contained in the
titration beaker.
Quinhydrone is formed from equimolecular proportions
of quinone and hydroquinone. When quinhydrone is dissolved
in aqueous solution, it dissociates, forming quinone and
hydroquinone:
Quinhydrone
quinone
+ hydroquinone
The equilibrium established between the dissociation products

may be indicated by the following equation:
Quinone
+ 2H + 2( -
hydroquinQne
The ratio of quinone to hydroquinone in acid 'solution is maintained constant by the presence of undissociated quinhydrone
in solution. When this condition obtains, the potential at the
electrode is a function of the hydrogen ion concentration of
the solution. In carrying out determinations, the potential
is measured against that of a calomel electrode, and the pH
value is calculated from the measured voltage in the same
manner as with the hydrogen electrode.
The potential of the quinhydrone electrode is higher than
that of the hydrogen electrode in the same solution. If a
saturated calomel electrode is used and the measurement is
carried out at 25C., the pH may be calculated from the equation:
0.453
- V
h
V'IS th e measure d vo1tage.
0.0591
= I
og I
H+ = PH were
289
Differences of 1 or 2C. in the working temperature do not

affect the results greatly. The pH may also be found from the
curve shOwing the relationship of pH to voltage for the quinhydrone electrode (Fig. 45).
The quinhydrone functions well in acid solution but not in
alkaline solutions. It can be used in alkaline solution where the
pH does not exceed 8. Alkaline solutions can be titrated, however, by adding a known excess of standard acid and titrating
the excess acid with an,alkali of known normality.
1\
pH
10
-0.5 -0.4 - 0..
- 0.2
- 0.1
Voltage
.. 0.1 .. O.l.
FIG. 45.-Graph showing the relation of pH to voltage measured with the

quinhydrone and calomel electrodes at 25C.
Exercise 90
Object.-To Determine the pH of Elixir of Iron, Quinine, and

Strychnine, Elixir of Pepsin, and Tincture of Aconite by Means
of the Quinhydrone Electrode.
Materials Required.-A potentiometric titration apparatus, without the
cylinder of hydrogen or the gas-washing bottles.
A plain platinum wire, foil, or gauze electrode.
50 to 100 mg. of quinhydrone.
50 cc. of each preparation.
Potassium biphthalate solution (see page 296).
290
Procedure.-l, Assemble the apparatus as in Exercise 87 (page 281),

replacing the hydrogen electrode with a platinum electrode. 'li'est the
accuracy of the apparatus against the standard biphthalate solution as
follows: Place sufficient of the biphthalate solution in the titration beaker
to cover the platinum electrode completely as well as the arm of the calomel
electrode. Stir about 5 mg. of crystalline quinhydrone into the biphthalate
solution. Mter 3 or 4 min. measure the voltage with the potentiometer.
The surface of the platinum electrode must be clean and bright.

It can be cleaned by immersing it in boiling nitric acid, washing
it in ammonium hydroxide and distilled water successively.
The pH of the biphthalate solution corresponding to the voltage
measured can be found from the graph (page 289). When
the apparatus is working properly, the pH of the biphthalate
mixture should be between 3.97 and 3.98.
2. Remove the titration beaker containing the biphthalate mixture, wash
the platinum electrode with distilled water, and flush a little Kel solution
through the side arm of the calomel electrode. Place about 50 cc. of the
preparation in the titration beaker and stir about 50 mg. of quinhydrone
into the elixir: Measure the voltage with the potentiometer.
Usually 2 or 3 min. are required to elapse after stirring in

the quinhydrone before the voltage becomes C6nstant. The pH
value corresponding to the voltage measured can be found from
the graph (page 289).
1. Name several pharmaceutical preparations of different types in which

the quinhydrone electrode can be used for measuring the
pH.
2. Explain how the end point in the titration of alkaloids
with acids might be determined potentiometrically.
The Glass Electrode.-It has been found that a

very thin membrane of special glass separating two
\.. solutions gives potentials which depend directly on
4~~ the hydrogen ion concentrations. The glass elec~rode with an trode (Fig. 46) consists of a glass electrode bulb and
mternal e l e - .
ment.
an mternal ref erence e1ectro de. Wh en t h e g1ass
membrane is mounted in a cell of the type, calomel
electrode, solution of known pH, glass membrane, solution of
unknown pH, calomel electrode, the voltage is a function of the
difference in hydrogen ion activity except for a small" asymmetry
Gl!::'
291
potential" due to the glass. A sensitive electrometer or an

amplifier of the vacuum tube type is required to measure the
voltages because the resistance of the glass diaphragm is so great
that the ordinary galvanometer is inaccurate.
A glass electrode pH meter of the type illustrated in Fig. 47
is suitable for most pharmaceutical work where the hydrogen
electrode or the quinhydrone electrode cannot be used. Instruments of this type are furnished with complete directions for
FIG. 47.-Glass eleotrode pH meter.
(Courteay Central Scientific Company.)
their preparation, operation, and maintenance. Under ordinary

conditions, pH values between 1 and 9.6 can be determined with
an accuracy of 0.1 pH unit. Above pH 9.6, buffer solutions
having approximately the same positive ion concentration as
the unknowns being tested should be used to calibrate the instrument. The glass electrode cannot be used to determine the pH
of alcoholic solutions. It can be used to determine the pH of
viscous and semisolid substances as well as colloidal and opaque
or colored solutions and solutions containing strong oxidizing or
reducing agents. Thus the pH of fats and oils, ointments, soaps,
emulsions, pastes, jellies, syrups, serums, infusions, cosmetics,
292
and many other pharmaceutical products can be determined by

means of the glass electrode.
Colorimetric Methods.-A number of colorimetric methods
for the approximate determination of hydrogen ion concentration
have been developed. The most generally used of these involve
the matching of the color obtained when a definite quantity of
indicator is ~dded to a definite volume of the liquid being tested
with that of pne of a series of standard buffer solutions differing
by about 0.2 pH unit. The volume of the unknown and of the
standard buffer solution must be the same, both of them must
contain the same amount of indicator solution, and the colors
must be matched by viewing through the same depth or thickness
of solution. Except in a few cases, the limit of accuracy attainable by the colorimetric method does not exceed 0.1 pH unit.
For convenience, the colorimetric method of determining
hydrogen ion concentration has been adopted for official purposes
and is to be employed where pigments or proteins present in the
solution do not vitiate its use. In these instances, the potentiometric method must be employed.
The principle of the colorimetric methodpepends upon the
assumption that an indicator in the same c,oncentration will
exhibit the same transition shade in different aque~us solutions
if the hydrogen ion concentrations of the solutions are the saine.
Having then a series of buffer solutions of definite hydrogen ion
concentrations, comparisons can be made betweerl the shades
of color produced in the buffer solutions and that produced by
the same indicator in the solution to be tested.
As previously stated, the Ka of an acid is affected by the
ionic strength of the solution, likewise the Ka of indicators is
also affected. This influences the color transition range of the
indicator. The electrometric method measures the activity
of the hydrogen ions (H+) and to obtain the same value by the
colorimetric method in solutions of various ionic strengths, a
correction, known as the salt-error correction, must be employed.
In addition, the kinds of ions present in solution and their
precipitating influence on the indicator affect the accuracy of
the colorimetric method. For official purposes, where the
complexity of the medicaments makes a simple application of
these corrections impossible, the corrections are ignored..
293
pH Indicators and Preparation of Their Solutions.-The

indicators used for colorimetric pH determinations are either
weakly acid or weakly basic. Most of the indicators used for
this purpose, however, such as the phthaleins and sulfonated
phthaleins, behave like weak acids.
The usual concentration of the indicator solution is 0.05 per
cent. From 0.1 to 0.2 cc. of the indicator solution is generally
used for 10 cc. of the liquid being examined.
Solutions 'of indicators of the basic type and of the phthaleins
are prepared by dissolving them in alcohol. In preparing solutions of indicators containing an acid group, ~his group must
first be neutralized with sodium hydroxide. The procedure
is as follows:
One-tenth (0.100) Gm. of the indicator is triturated in an agate
mortar with the volume of 0.05 N sodium hydroxide specified
in the following table, or with its equivalent of 0.02 N sodium
hydroxide. When the indicat~r has dissolved, the solution is
diluted with carbon-dioxide-free distilled water to make 200 cc.
(0.05 per cent).
The solutions should be kept in stoppered bottles, and protected from light.
TABLE
Indicator
XL.-pH
pH range
INDICATORS
MolecuJar
weight
Color change
Solvent
-Methyl yellow .......... .... 2.9

Bromphenol blue ........ ... 3.0
Methyl red ............... .4.2
Bromcresol purple ........... 5.2
Bromthymol blue ........... 6.0
Phenol red .............. .. 6.8
Thymol blue .. ............ 8.0
Thymolphthalein ..... , . .... 9.3
to
to
to
to
to
to
to
to
4.0
4.6
6.3
6.8
7.6
8.4
9.6
10.5
225
669
269
540
624
354
466
430
Red-yellow
Alcohol
Yellow-blue
3.0 cc. 0.05 N
Red-yellow
7.4 cc. 0.05 N
Yellow-purple 3.7 cc. 0.05 N
Yellow-blue
3.2 cc~ 0.05 N
Yellow-red
5.7 cc. 0.05 N
Yellow-blue
4.3 cc. 0.05 N
Colorless-blue
Alcobol
NaOH
NaOH
NaOH
NaOH
NaOH
NaOH
Solutions Used in the Preparation of Buffer Solutions.Fifth molar hydrochloric acid and 0.2 M sodium hydroxide
(carbonate-free) are prepared and standardized according to
the directions given under Standard Solutions.
294

TABLJ!J XLL-BuFFER MIXTURES OF CLARK AND
Luns
HCI-KCI Mixtures
pH
1.1
1.2
1.3
1.4
1.5
1.6
1.7
1.8
1.9
2.0
2.1
2.2
0.2 M HOI, cc.
0.2 M KOI,
CC.
5.44
24.90
40.32
52.60
62.36
70.06
76.24
81.14
85.02
88.10
90.64
92.48
94.56
75.10
59.68
47.40
37.64
29.90
23.76
18.86
14.98
11.90
9.46
7.52
Dilute to, cc.
200
200
200
200
200
200
200
200
200
200
200
200
Phthalate-Hel Mixture
pH
0.2 M KH Phthalate. co.
2.2
2.4
2.6
2.8
3.0
3.2
3.4
3.6
3.8
50
60
60
50
50
50
50
50
50
0.2 MHCI
46.60
39.60
33.00
26.50
20.40
14.80
9.95
6.00
2.65
J:?ilute to,
00.
200
200
200
;ZOO
200
200
200
200
200
Phthalate-NaOH Mixtures
pH
0.2 M KH Phtha}ate. cc.
4.0
4.2
4.4
4.6
4.8
5.0
5.2
5.4
5.6
5.8
6.0
6.2
50
50
50
50
50
50
50
50
50
50
50
50
0.2 M NaOH, cc.
0.40
3.65
7.35
12.00
17,50
23.65
29.75
35.25
39.70
43.10
45.40
47.00
Dilute to. ce.
200
200
200
200
200
200
200
200
200
200
200
200

TAllLE XLI.-BuFFER MIXTURES OF CLARK AND
295
LUBS.-(Continued)
KH.PO.-NaOH Mixtures
pH
5.8
6.0
6.2
6.4
6.6
6.8
7.0
1.2
7.4
7.6
7.8
8.0
0.2 M KH.PO., cc.
50
50
50
50
50
50
50
50
50
50
50
50
0.2 M NaOH. cc.
3.66
5.64
8.55
12.60
11.14
23.60
29.54
34.90
39.34
42.14
45.17
46.85
Dilute to,
CC.
200
200
200
200
200
200
200
200
200
200
200
200
Borio Acid, KCI-NaOH Mixtures
pH
7.8
8.0
8.2
8.4
8.6
8.8
9.0
9.2
9.4
9.0
9.8
10.0
0.2 M H,BO" 0.2 M Kel,

00.
50
50
50
50
50
50
50
50
50
50
50
50
0.2 M NaOH, ce.
2.65
4.00
5.90
8.55
12.00
16.40
21.40
26.70
32.00
36.85
40.80
43.90
Dilute to, cc.
200
200
200
200
200
200
200
200
200
200
200
200
1. Potassium Biphthalate Solution.-Dissolve 40.836 Gm. of

reagent potassium biphthalate in distilled water and dilute to
1,000 ee. to prepare a 0.2 M solution for use in preparing the
buffer solutions.
2. MonopotassiumPhosphate Solulion.-Dissolve 27.240 Gm. of
reagent monopotassium phosphate in distilled' water and dilu.te
to 1,000 ce. to make a 0.2 M solution for use in preparing the
buffer solutions.
3. Boric Acid and Potassium Chloride Solution.-Dissolve
12.404 Gm. of reagent boric acid and 14.912 Gm. of reagent
296
potassium chloride in distilled water and dilute to 1/000 cc.

to prepare a 0.2 M solution of these mixed salts.
'
4. Potassium Chloride Solution.-Dissolve 14.912 Gm. reagent
potassium chloride in distilled water and dilute to 1/000 cc. to
make a 0.2 M solution for use in preparing the buffer solutions.
5. Potassium Biphthalate Solution for Hydrogen Electrode.Solution (1) diluted with three volumes of distilled water at
20C. prepares 0.05 M potassium biphthalate solution for checking the hydrogen electrode.
Directions for the pH Measurements.-In the determination
of an unknown pH value, a suitable indicator must first be found ..
Only indicators which show an intermediate color between the
extreme acid and alkaline colors can be used.
The first step in the choice of a suitable indicator is the determination of the approximate pH value of the solution under investigation. A few simple tests will usually supply the necessary
information. Add a drop or two of phenolphthalein T.S. to a
small portion of the solution. If the indicator remains colorless,
the pH of the solution is less than 8.4. A second test is conducted in the same manner/ using methyl o.:range T.S. as the
indicator. If the solution assumes the alkaline color (yellow),
the pH of the solution is greater than 4.4 and liesl somewhere
between 4.4 and 8.0. A few more tests with methyl red (pH
interval 4.2 to 6.3), bromthymol blue (6.0 to 7.6), and phenol
red (6.8 to 8.4) will give a close enough approximation of the
pH value to show which indicator may be successfully used in
the determination.
Instead of testing small amounts of the liquid with indicator
solutions, the spot method, using indicator papers, may be
substituted. Also, by using a universal indicator in place of
the several indicator solutions suggested above, some time may
be saved. A so-called universal or mixed indicator of this type
can be prepared as follows: Dissolve 0.1 Gm. of phenolphthalein,
0.2 Gm. of methyl red, 0.3 Gm. of dimethyl-aminoazobenzene,
OA Gm. of bromthymol blue, and 0.5 Gm. of thymol blue in
500 cc. of absolute alcohol and then add sufficient sodium
hydroxide to produce a yellow color. When 0.1 to 0.5 cc. of this
mixed indicator is added to an unknown solution, the color produced indicates the approximate pH of the solution as follows:
297
Red color = pH 2; orange color = pH 4; yellow color = pH 6;

green color = pH 8; and blue color = pH 10.
When the approximate pH value has been determined and a
suitable indicator agreed upon, a 3, 5, or 10 ce. portion of the
unknown solution (depending upon the amount of the liquid
available), is trap.sferred to a hard, resistant-glass test tube
approximately 15 cm. long and 1.5 cm. 0.5 mm. bore, and a
measured amount of the indicator solution added. As a rule,
0.10 to 0.20 cc. of a 0.05 per cent indicator solution, added from
a 1 cc. pipette, graduated to 0.01 cc. per 10 cc. of the solution
being tested, constitutes a proper indicator. concentration.
Transfer from 4 to 6 portions of the buff~r solutions, the pH
values of which overlap that of the unknown solution, to test
tubes and treat in exactly the same way as the s~lution being
analyzed. The same amounts of indicator must be added to
the unknown and to the buffer solutions. It is also essential
that the test tubes used be of the quality and type already
indicated. The color of the unknown solution is then compared
with the colors of the buffer solutions and the pH value of the
solution thus determined.
In judging the colors, observe them against a white background
with the light transmitted through the whole length of the tube.
A suitable colorimeter may also be used, although it is not necessary in routine work. A sufficient number of reference solutions
must be taken so that the color of the unknown falls between
two of the series, differing by not more than 0.20 pH. The pH
of the unknown can thus be easily approximated to within 0.1
and with practice to 0.05. With buffer solutions differing by
0.1 pH unit or less, the experimental error can be reduced to
about 0.02 pH.
Other colorimetric methods capable of the same or a greater
degree of accuracy than that officially required may be employed
at the discretion of the operator.
The dissociation constants of'indicators hold for aqueous solutions only. Alcohol changes this value and therefore for alcoholic
solutions, a correction factor must be used or the potentiometric
method employed.
Color standards are prepared by placing 10 cc. of a buffer
solution of the proper pH in each of nine Pyrex tubes, adding
298
QUA.NTITATIVE PHARMACEUTICAL CHEMISTRY
0.5 cc. of the indicator solution to each of them, and then sealing
the tubes. Thus in preparing color standards for phenol red
10 cc. of nine buffer mixtures having pH values of 6.8, 7.0, 7.2,
7.4, 7.6, 7.8, 8.0, 8.2, and 8.4, respectively, are placed in separate
tubes, 0.5 cc. of 0.02 per cent phenol req. indicator solution is
added to each tube, and the tubes are sealed. Each tube is
then labeled with the respective pH value corresponding to
that of the buffer solution which it contains. The tube marked
pH 6.8 will exhibit the yellow color and that marked pH 8.4 will
exhibit the red color of phenol
red, and the tubes with intermediate pH values will show
gradations of color between
yellow and red.
Reliable color standards
covering the pH range 1.2 to
10.4 for all of the required
indicators, guaranteed for one
year, can be purchased. It is
unnecessary to secure color
standards for all of the indicat ors cove ring the range
FIG. 48.-Block comparator.
pH = 1.2 to 10.4 unless determinations of pH on a variety of products of widely divergent
hydrogen ion concentration are to be made. Thus if a solution
has a pH falling between 6 and 8.4, color standards for bromthymol blue and cresol red are all that is necessary.
Color Comparators.-Some type of color comparator must be
used to match the color of the unknown with the color standards.
This is especially true if the sample under examination is at all
turbid or colored. Of the numerous devices available, the
LaMotte block comparator (Fig. 48) is 1>est suited for use with
the equipment herein described. This comparator consists of
anyone set of color standards such as bromthymol blue; four
test tubes graduated at 10 cc. and of the same bore and wall thickness as the color standard tubes; and a 50 cc. bottle of the corresponding indicator solution; a tube of distilled water; and a
pipette contained in a wooden case. The top of this case is so
designed that it may be removed and used as a comparator
block.
299
The block comparator is used as follows: Assume that a preliminary test made with various indicators or a mixed indICator
shows that the solution being tested has a pH falling between
pH 6.0 and 7.5. A set of bromthymol blue color standards and
indicator would then serve for the determination. To make the
determination, remove the top of the comparator case (Fig. 48),
and use it as a comparator block. The six holes back of the slots
in the side of this block are desigB
A
c
nated B, A, C and E, D, F, respectively, in Fig. 49.
Fill three of the test tubes to the
10 cc. graduation mark with the
liquid to be tested and place them in
the holes marked B, A, and C. Add
0.5 cc. of bromthymol blue to the
F
D
tube in A and shake the tube well to
.E
mix the contents. Place the tube con- FIG. 49.-Positions of tubes
in the block comparator.
taining distilled water in the hole
marked D, and two of the tubes containing color standards
differing only by 0.2 pH, e.g., 6.8 and 7.0, in the holes E
and F, respectively. Place the comparator block before a
window or other source of light, observe the three pairs
of tubes through the slots, and change the color standards
in E and F, if necessary, until the color of the central pair exactly
matches the color in one of the other pairs or until the color of
the central pair lies between that of the pairs on either side.
If the color of the solution in the central pair matches that of one
of the other pairs, the pH is read off on the color standard with
which the match is obtained. If, however, the color of the solution in the central pair of tubes lies between the colors of the
pairs on either side, the average pH of the latter is taken as the
pH of the sample; e.g., if it lies between 7.0 and 7.2, the value
is taken as 7.1.
The LaMotte roulette comparator (Fig. 50) is designed for
rapid pH determinations with artificial illumination. This comparator consists of a stationary base and metal band, and a
wooden drum which revolves inside the band on ball bearings.
Illumination is provided by a 40-watt Mazda lamp attached in
the center of the base. A piece of "Dalite" glass is placed in the
000
000
300
back of the block between the three test tubes and the color
standards, and a piece of etched glass is placed on the outside
of the block directly over the three slots. This contrivance provides standard conditions of illumination at all times. Ampoules
of distilled water, bottles of indicator solution, graduated test
tubes, and pipettes are a part of the comparator equipment.
Any three sets of color standards such as chlorphenol red,
pH 5.2 to 6.8; bromthymol blue, pH 6.0 to 7.6; and phenol
red, pH 6.8 to 8.4 are placed in alternate holes in the revolving
drum in the order of their pH. Tubes of the same bore filled
with distilled water are then placed in the vacant holes. There
FIG.
SO.-The roulette comparator with accessory equipment.
will then be :3, tube of distilled water beside each tube of color
standard. If the pH of the solution to be tested falls somewhere
within the range of bromthymol blue, pH = 6.0 to 7.6 as determined by preliminary tests (see page 296), fill three of the test
tubes to the 10 cc. graduation mark with the solution and place
them in the three holes in the block. Add 0.5 cc. of bromthymol
blue to the middle tube and mix it in well by shaking. Turn on
the light and revolve the drum until the bromthymol blue color
standards are directly behind the test sample. While looking
through the slots in the block, gradually shift the standards by
rotating the drum until the color through the test sample exactly
matches that of one of the tubes on either side of it or lies between
them. The pH value may then be read off directly from the color
standards in the same manner as with the block comparator.
301
Exercise 91
Object.-To Determine the pH of Solution of Epinephrine

Hydrochloride, and Syrup of Hydriodic Acid.
Materials Required.-A set of color standards.
Indicator solutions corresponding to the color standards.
A color comparator.
40 cc. of solution of epinephrine hydrochloride, and syrup of hydriodic
acid.
Procedure.-Determine the pH of each of the respective preparations
by the method described for either the block or roulette comparator. Calculate the hydrogen ion concentration of each preparation.
1. Enumerate several instances in the description of official products
where such terms as slightly acid, alkaline to litmus, etc., could be replaced
by definite limits of pH to advantage.
2. What advantages does the colorimetric method have that are not
possessed by the hydrogen electrode method of hydrogen ion determination?
3. Name several classes of pharmaceutical preparations of which the pH
cannot be determined colorimetrically.
4. Ascertain the official pH requirements of water, distilled water,
redistilled water, sterilized distilled water, and reagent potassium biphosphate. How may the pH limits be determined?
6. What use is made of pH in the determination of the purity of oxygen
and nitrogen monoxide? (See U.S.P.)
6. It is required in the official standards that fluidextract of aconite be
adjusted to a pH of 2.75 to 3.25 and that tincture of aconite be adjusted
to a pH of 2.8 to 3.2. If one of these preparations had a pH of 4.0, how
would you proceed to adjust it to the official requirements?
Hydrogen Ion Concentration (pH) of Some Official Substances.

The hydrogen ion concentrations, expressed as pH, of the substances in the following table are not intended to be construed as a
means for the determination of the purity of these substances.
In practice, variations from these figures may frequently be
found, as a slight excess of acid or base is, in many instances,
desirable and even necessary to insure stability and other qualities
in connection with the use of these substances.
When only one figure is given in the table it represents the
theoretical pH, or the one generally agreed upon in the literature.
For the majority of the alkali and alkaline earth salts, an approximate range is given within which the pH of these substances, as
302
QUANTITATIVE PHARMAcEUTICAL CHEMISTRY

TABI~E
XLII.-pH
OF SOME OFFICIAL SUBSTANCES
Substance
Concentration
Acid, acetic ......................... .

Acid, benzoic ................... .
Acid, boric ...................... , .. .
Acid, citric ......... , , ..... , .. , , .. , .. .
Acid, hydriodic ...... : ... , ........... .
Acid, hydrbchloric .. , ............ , ... .
Acid, hypo phosphorous . , , ........ , ... .
Acid, lactic .............. , ...... , ... .
Acid, nitric., ...... , .... ' .. , ...... , .. .
Acid, phosphoric .......... , ...... , .. .
Acid, salicylic ... ~ ......... , ...... , .. .
Acid, sulfuric ...... ,., .... , ...... , .. .
.Acid, tartaric ....................... .
Acid, trichloroacetic, ....... , ......... ,
Aluin (ammonium). . .. . ... , ...... , .. .
Alum (potassium) .......... , ....... , ..
Ammonium bromide ............... , .. .
Ammonium chloride ... , ...... , ....... .
Ammonia water.' ................. .
.t\pomorphine h:ydrochloride ....... , , .. .
Arpphenamine ............. , ......... .
Atropine ........ ~ ................... .
Atropine sulfate .... , ...... , .... , . , .. .
Caffeine citrated .. , . , ...... , ......... .
Caffeine with sodium benzoate ....... , ..
Calcium bromide ..................... ,
Calcium chloride ........... , ...... , .. .
Calcium hydroxide ....... , ........ , .. .
Calcium lactate ........... , , ..... , , .. .
Cinchonidine sulfate ............ , .. , .. .
Cocaine hydrochloride ..... , , . , .... , .. .
Codeine ............ , .. " ........ ,.,.
Codeine phosphate. .. ! . . .
Codeine sulfate .... , ........ , ....... , ,
Emetine hydrochloride ....... , ... , , ., .
Ephedrine ........................ , .. .
Ephedrine hydrochloride .. , , , ...... , . , .
Homatropine hydro bromide ... , ... , ... .
Magma magnesia ................. , .. .
Magnesium sulfate ....... , ........... .
Morphine ~ulfate. . . . .. .,. . ...... , ..
Physostigmine salicylate ........... , ..
Pilocarpine nitrate ................ , .. .
0.1 M
Saturated fjolution
0.1 M
0.1 M
0.1 M
0.1 M
0.1 M
0.1 M
0.1 M
0.1 M
Saturated solution
0.05 M
0.1 M
0.1 M
0.05M
0.1 M
0.1 M
0.1 M
0.1 M
1 in 300
1 in 20
Saturated solution
1 in 100
1 in 25
1 in 25
0.2 M
0.2 M
Saturated solution
1 in 25
Saturated solution
0.1 M
Saturated solution
0.1 M
0.1 M
1 in 50
1 in 200
1 in 200
1 in 100
0.2 M
0.1 M
1 in 200
1 in 100
pH
2.9
2.8
5.1
2.1
1.0
1.0
1.5
2.4
1.1
1.5
2.4
1.2
1.9
1.2
4.6
4.2
4.6
4.6
11.3
4.8
3.0
\9.5
5.4
2.3
7.4
8.0
7.0
6.5 to 7.5
13.5
6.0 to 7.0
6.4
4.5
9.8
4.5
5.0
5.6
to
10.8
5.1
4.4
10.6
6.0to7.0
4.8
5.8
4.8

TABLE
XLIL-pH
OF SOME OFFICIAL
Substance
Potassium acetate .................... .
Potassium bicarbonate ..... , ..... : .... .
Potassium bromide ... ,'. , , ............ .
Potassium carbonate ...... , , ......... .
Potassium hydroxide ................. .
Potassium iodide .............. , ...... .
Potassium nitrate .................... .
Potassium and sodium tartrate ... , .... .
Procaine hydrochloride ............... .
Quinidine sulfate ... , ....... , ......... .
Quinine ............................. .
Quinine bisulfate ..................... .
Quinine dihydrochloride .. , , .......... .
Quinine hydrobromide ................ .
Quinine hydrochloride ............... , .
Quinine sulfate ..... , ..... , ........ , :.
Quinine and urea hydrochloride ........ .
Sodium acetate ...................... .
Sodium benzoate ..................... .
Sodium bicarbonate .................. .
Sodium biphosphate .......... , ....... .
Sollium borate .... ,., , ...... , ....... :.
Sodium bromide, . , ....... , .......... .
Sodium cacodylate, ........ , ...... , .. .
Sodium carbonate, ..... , , ........ , ., ..
SQ.dium chloride ... , ................. .
Sodium hydroxide .................... .
Sodium iodide ....................... .
Sodium phosphate (dibasic) ........... .
Sodium salicylate. , .................. .
Sodium sulfate ............ , .......... .
Sodium thiosulfate ..... , ............. .
Soluble barbitaL, .................... .
Strychnine nitrate ........... .' ...... , ..
Strychnine sulfate .................. , ..
Theobromine with sodium salicylate ... .
303
SUBSTANCES.-(Continued)
Concentration
pH
0.1 M
9.7
0.1 M
8,2
0.2M
6,5 to 8.0
0.1 M
11..6
0.1 M
13.5
0.2M
.7.0 to 9.0
0.2M
6.5 to 7.5
0.2 M
7.0to8.0
0.1 M
6,0
1 in 200
6.4
Saturated solution
8.8
1 in 25
3.5
1 in 25
2.6
1 in 25
6.4
1 in 25
6.'4
Saturatt;ld solution
6.2
1 in 20
3.1
0,1 M
9.7
0.1 M
8.0
0.1 M
8.2
0.1 M
4.5
9.2'
0.1 M
0.2 M
6.5 to 8.0
0.1 M
7.8
0.1 M
11,6
0.2M
6.7to 7.3
0.1 M
13.5
0.2 M'
8.0 to 9.5
0.1 M
9.5
0.2 M
5.0 to 6.0
0.2 M
6,0 to 7.5
0.2M
6.5 to 8.0
0.1 M
9.4
1 in 250
5.7
1 in 100
5.5
1 in 100
10.3
they usually occur on the market, will falL Some deviations from
these values may, however, be expected, as the presence of even
a very slight excess of base or acid in these salts, or of carbon
dioxide in their solutions, exercises a pronounced influence upon
the hydrogen ion concentration.
304

References
1. BRITTON, "Hydrogen Ions," Chapman and Hall, Ltd., London, 1932.

2. CLARK, "The Determination of Hydrogen Ions," Williams & Wilkins
Press, Baltimore.
3. KOLTHOFF and FURMAN, "Potentiometric Titrations," John Wiley &
4. Leeds Northrup and Company Bulletins, Leeds Northrup and Company,
Philadelphia.
CHAPTER XIX
ELECTROLYTIC METHODS
Quantitative analysis by means of electrolysis, or electroanalysis, as it is more generally called, is usually restricted to the
determination of metals. This method of analysis is based upon
the fact that an electrical current passed through a solution of
the salt of a metal causes the deposition of the metal, usually
in the elemental state, upon one of the electrodes. The electrolytic method may be applied to a number of official substances,
namely, mercury and its salts and the salts of silver, copper, and
zinc.
Electrical Units and Fundamental Laws.-The unit of current
is the ampere, that of resistance is the ohm, and that of difference
of potential or el~ctromotive force is the voU. The ampere is
the strength of current which when passed through a solution of
silver nitrate under certain standard conditions will deposit 0.001118
Gm. of silver per second. The ohm is defined as the resistance
offered to an unvarying electric current by a column of mercury
106.3 cm. long and 1 sq. mm. in cross-section at 00. The voU
is the electromotive force necessary to force a current of one ampere
through a resistance of one ohm. The relationship between the
ampere, volt, and ohm is expressed in Ohm's law, viz.: The
strength of an electric current flowing through a conductor is directly
proportional to the difference of potential between the ends of the
conductor and inversely proportional to its resistance. If C represents the strength of the current in amperes, E the difference of
potential in volts, and R the resistance in ohms, Ohm's law may
be formulated as follows:
E
C = Ii
or, transposed,
E = OR
- The calculation of the amount of metal which will be deposited
during electrolysis in a given period of time is founded on Faraday's laws, which may be stated as follows:
305
306
1. The mass of any substance deposited at an electrode from a

solution of electrolyte is proportional to the quantity of electricity
which passes through the solution.
2. The amounts of different substances liberated at the electrol'1es,
when the same quantity of electricity passes through solutions of
different electrolytes, are proportional to the chemically equivalent
weights of the substances.
From the first law of Faraday it follows that the weight of
substance deposited from solution during a given p_eriod of time
will be directly proportional to the current strength or amperagtt,
and under a given amperage, will be directly proportional to the
time.
The unit quantity of electricity is the cou.lomb. It may be
defined as the amount of electricity which passes through a conductor
when one ampere flows for one second. It has been found by
experiment that 96,500 coulombs of electricity are required to
liberate a gram-equivalent weight of any substance at an electrode. The electrochemical equivalent of an element or group of
elements is the weight in grams deposited at an electrode by the
passage of one coulomb of electricity. The electrochemical
equivalents are proportional to the chemical equiv~lents. Thus
96,500 coulombs of electricity are capable of deposit\ng Hl1
= 1.008 G111. hydrogen. Therefore, 1 coulomb <!an liberate
1.008 -;- 96,500 = 0.0000104463 Gm. hydrogen. This same
quantity of electricity will liberate 35.45 X 0.0000194463 =
0.00036749 Gm. chlorine, and 107.88 X 0.0000104463 = 0.001118
Gm. silver, and 96,500 coulombs of electricity will deposit
Ag/1 = 107.88 Gm. silver from a solution of a silver salt; Cu/2
= 31.785 Gm. copper from a solution of a cupric salt; and Fe/3
= 18.61 Gm. iron from a solution of a ferric salt. The quantity,
96,500 coulombs, is thus observed to be a unit of quantity in
electrochemical measurements. This unit of quantity is known
as the Faraday, and it is commonly represented by the letter F.
Theory.-Those substances which conduct an electric current
when dissolved in aqueous solution are termed electrolytes.
Electrolytic determinations are based upon the fact that when
a direct current of electricity is passed through a solution of an
electrolyte, some of the negatively charged ions are oxidized to
the elemental state, and some of the positively charged ions are
307
reduced to the elemental state. This decomposition of an

electrolyte in solution is, known as electrolysis, and when the
de.composition of the electrolyte is carried out for purposes of
analysis, the process is called electro-analysis. The conductor
b:y which the current enters the solution of electrolyte is called
the anode, and that'by which it leaves the solution is known as
the cathode. The anode and cathode are generally spoken of
a:s positive and negative electrodes, respectively, or simply as
electrodes. The source of current charges the positive electrode
(anode) with positive electricity, and this anode attracts the
negatively charged ions (anions), but it repels the posith:ely
charged ions (cations); the latter are attracted by the negatively
charged cathode which repels the negatively charged anions
forcing them toward the anode. Since the positively charged
ions gain an electron for each positive charge which they carry
when they come in contact with the cathode, they are reduced;
and since the negatively charged ions lose an electron for each
negative charge which they carry when they come in contact
with the anode, they are oxidized. The respective reductions
and oxidations never take place in the mass of the solution of
electrolyte but only at the electrodes, reduction occurring at the
cathode and oxidation occurring at the anode. Simple metallic
ions (cations) such as copper, mercury, silver, and zinc travel to
the cathode where they are reduced to the metallic state. The
simple anions, such as chlorine, bromine, or iodine travel to
the anode and are oxidized to the molecular condition, while
solutions containing hydroxide, sulfate, or nitrate ions liberate
oxygen at the anode. The halogen or oxygen liberated may
escape from the solution or it may act directly or indirectly as an
oxidizing agent.
The various anions are not all oxidized with the saine ease,
their relative order being sulfide, iodide, bromide, chloride, and
sulfate, where the first named is the most readily oxidized.
Likewise, the different cations are not all reduced with the same
ease, their relative order being auric, platinic, silver, mercurous,
cupric, arsenous, bismuth, liydrogen, and lead, where the first
named is most readily reduced. The electrolytic method of
analysis is, therefore, a type of oxidation-reduction analysis,
since the oxidation of one species of ion always takes place at the
308
anode simultaneously with the reduction of another species of

ion at the cathode.
The minimum voltage necessary to separate ions from the charges
which they carry is known as the decomposition voltage. The
decomposition voltages of the ions of official substances which
can be assayed by this method are given in the following table:
TABLE XLIII.-DECOMPOSITION VOLTAGES
Ion
Metal
Zn++ ........... .
Fe++ ............. .
Sn++ ...... . .......
Pb++ ............ .
H+ ................
CU++ .. ....... .... .
Hg++ ..............
Ag+ ...............
OH- ..............
Zn
Fe
. Sn
Pb
. H 2 (Pt)
. Cu
. Hg
. Ag
. 02CPt)
1-................. . 12 (Pt)
Br- ... .
Cl- .... .
Br2(Pt)
C1 2 (Pt)
Decomposition
voltage
-0.7581 to 0.80
-0.441 to0.44
-0.136 to 0.14
- 0 . 122 to O. 12
0.0
toO.O
+0.3448 to 0.34
+0.7986 to 0.80
+0.7995 to 0.80
+0.3957 to 0.40
+0.5357 to 0.54
+ 1. 0659 to 1. 066
+ 1. 3594 to 1. 36
CPt) = platinum electrode.
Since during electrolysis deposition of the ions occurs at both

electrodes, the voltage required to liberate the ions at both the
anode and cathode must be considered. For example, when
mercury ions from a solution of mercuric iodide are reduced to
metallic mercury, the decomposition voltage for the mercury ion
is +0.80 while that for the iodide ion is +0.53. Since these
voltages have the same sign, their sum, when added algebraically,
is 0.80 - 0.53 = 0.27. The minimum voltage at which mercury
and iodine can be liberated from mercuric iodide solution is
therefore, 0.27 volt. If zinc chloride is subjected to electrolysis, the respective decomposition voltages for the ions being
- 0.76 and
1.36, the decomposition voltage necessary is equal
to their algebraic sum, 2.12 volts.
The difference in the decomposition voltages of mercuric iodide
and zinc chloride solutions make it possible to separate mercury
from zinc in a solution containing these salts by maintaining the
309
voltage below 2.12 and above 0.27. The greater the difference is
between the decomposition voltages of two metallic ions the easier
it is to separate them by electrolysis. In general, it may be said,
that the decomposition voltages of two substances must differ
by at least 0.2 to 0.3 volt in order to permit their separation by
electrolysis.
Electrolytic methods of assay are not so simple as is implied
in the foregoing examples, since numerous factors complicate
conditions, viz., resistance of the solution, polarization, overvoltage, reactions at the electrodes, the nature of the solutions
used, the current density, the rate of stirring, the shape and size
FIG. 51.-Gauze anode.
FIG. 52.-Spiral anode.
FIG. 53.-Spiral anode.
of the electrodes, and the temperature. A discussion of all of

these factors is beyond the scope of the present workj those who
desire a more comprehensive knowledge of them should consult
a treatise on physical chemistry.
Apparatus.-The most satisfactory source of current for
electrolytic assays is that furnished by a lead storage cell or a
number of such cells combined into a battery. Each cell should
supply an electromotive force of about 2 volts so that the connection in series of two or more cells will make it possible to
secure a voltage range of from 2 to 12 volts. The use of the
lighting current is possible provided it is a direct current, but
since the ordinary service voltage is usually 110 to 125 volts,
the voltage must be greatly reduced. An electrical switch is
necessary to make and break the current. The potential difference between the electrodes is regulated by means of a resistance
310
device. A rheostat may be employed, but the ordinary slidingcontact resistance coil is convenient and satisfactory for most
purposes. The difference of potential between the electrodes
is measured by means of a voltmeter. The amount of current
flowing through the electrolytic circuit is measured by an
ammeter. The voltmeter and ammeter employed should be
designed for a limited range of measurement so that they may
be read easily and accurately.
The anode employed in electrolytic determinations usually
consists of a spiral, gauze, or disk of platinum of such construction
that it can be attached to a stirrer. Platinum is employed
FIG.
54.-Platinum
cathode.
dish
FIG. 55.-Platinum
foil cathode.
FIG. 56.-Platinum
gauze cathode.
because it is very resistant to the chemical action of the anions.

Some of the commonly used forms of anodes are illustrated in
Figs. 51, 52, and 53. Although any rotating device may be
employed, a friction drive motor stirrer is very convenient, since
the number of revolutions of the anode per minute can be easily
adjusted.
The material of which the cathode is made depends upon the
metal which is to be deposited upon it. A platinum dish of
about 200 cc. capacity serves well as cathode, (Fig. 54) in all
of the electrolytic assays of official substances except that of
zinc. In the assay of mercury and its salts and preparations,
however, a mercury cathode cup is recommended. The mercury
cathode cup consists of a glass vessel about 7.5 cm. in height
and 4 cm. in diameter with a piece of platinum wire fused into
and through the bottom of the cup and containing about 50 Gm.
FIG.
311
57.-Diagram of connections for electrolytic apparatus. a, anode; c.

cathode. A, ammeter; B, battery; V, voltmeter; R, rheostat.
FIG.
58.-Rotating anode and stirring device.
312
of mercury. The platinum wire must extend through th~ glass

wall to make contact with the mercury. In the determination
of zinc electrolytically, either a nickel dish of about 175 cc.
capacity or a platinum dish upon which a thin layer of copper or
silver has been deposited may be employed as the cathode. The
platinum dish type of cathode can advantageously be replaced
by a hollow cylinder of platinum gauze or foil (Figs. 55 and 56).
These types of cathode are more efficient than the platinum dish
cathode because they present a large surface all parts of which
receive the deposit equally, and because they allow free circulation of the electrolyte. They are also more economical, since
they are light in weight.
The method of connecting up the apparatus is shown in the
diagram (Fig. 57).
A convenient rotating anode and stirring device is illustrated
in Fig. 58.
Immediately before starting an assay, both the anode and
cathode should be cleaned by immer~ing them in a hot solution
consisting of equal parts of concentrated nitric acid and distilled
water. They should then be thoroughly rinsed with distilled
water and ignited to a dull red heat over the blue flame of a gas
burner. This treatment removes any grease or othe~ impurity
present and leaves a clean electrode surface which is essential,
particularly with reference to the cathode, to the production of a
firm, even deposition of metal.
1. Define the following terms: ampere, ohm, volt, coulomb, Faraday,
anode, and cathode.
2. State Ohm's law and Faraday's laws.
3. Show by means of a diagram how the electrolytic apparatus is connected in the electrolytic assay of mercury when a mercury cathode cup
is used.
4. Calculate the decomposition voltage of mercuric chloride.
5. An electric current is passed simultaneously through a series of three
electrolytic cells containing water acidulated with sulfuric acid, silver
nitrate in sodium cyanide solution, and zinc sulfate in sodium hydroxide
solution, respectively. How much silver and how much zinc will be deposited if 0.504 Gm. of hydrogen gas measured at OC. and 760 mm. is liberated
from the water?
313
Exercise 92
Object.-To Assay Copper 8ulfate.

Materials Required.-An electrolytic apparatus.
1 Gm. of copper sulfate.
Hydrogen sulfide'r.S.
25 cc. of alcohol.
25 cc. of ether.
15 cc. of concentrated nitric acid.
Procedure.-1. Dissolve from 0.75 to 1.0 Gm. of the salt, accurately
weighed, in about 100 cc. of distilled water in a weighed platinum dish, add
1 cc. of diluted sulfuric acid, rotate the anode about 500 revolutions per
minute and pass the current, gradually increasing it to 2.5 amperes and from
5 to 7 volts. Occasionally add a little distilled water to the electrolyte to
keep the deposit of copper covered.
When the cupric sulfate is dissolved in water it ionizes:

Cu80c-~Cu++
+ 80
4- -
The copper ions migrate to the cathode where each copper ion
gains two electrons and is reduced to metallic copper. The
sulfate ions migrate to the anode where they react with hydroxyl
ions from the water, forming molecular oxygen and sulfuric
acid:
Each sulfate ion causes two atoms of oxygen combined as

hydroxyl ion to lose an electron and become oxidized to molecular
oxygen. The oxygen is given off at the anode and the hydrogen
ions remaining from the hydroxyl ions unite with the sulfate
ions to form H 280 4 If the current employed in the electrolysis
is too strong, however, some sulfate ions may also be discharged:
80 4- -
+ 80 ----+280 + O
3-
The 80 a formed then reacts with water to form H 280 4 :

80 3
+H
0--+H 280 4
The diluted sulfuric acid is added because it ionizes strongly

in solution and thereby increases the conductivity of the electrolyte and decreases the resistance of the solution to the passage
of current. It also supplies a high concentration of hydrogen
314
ions which tend to prevent the deposition of those metals which

are above hydrogen in the electromotive series and to prevent the
oxidation of the deposited copper to the oxide. If the electrolysis
of copper sulfate solution is carried out without the addition of
sulfuric acid, the copper deposit is spongy and non-adherent,
but when sulfuric acid is added, the deposit is compact and
adherent.
It has been found by experiment that vigorous stirring of the
electrolyte greatly reduces the time required to carry out an
electrolytic determination, since it brings the ions in contact with
the electrodes. The rotating anode serves as a stirrer.
Small amounts of distilled water are added from time to time to
cover and thereby prevent the oxidation of the freshly deposited
copper by the atmospheric oxygen.
2. When the copper is all deposited (usually in about 20 min., to be
ascertained by withdrawing a little of the electrolyte and testing it with
hydrogen sulfide T.S.), stop the rotation, and wash the deposit with distilled water by means of a siphon without interrupting the current until the
current drops to nearly zero.
The complete deposition of the copper is indicated by the

disappearance of the characteristic color from thel solution.
When the solution is colorless, transfer a drop of it to a depression
in a porcelain spot plate by means of a glass stirrin~ rod and
add a drop of hydrogen sulfide T .S. If the copper has not all
been deposited, a brownish-black precipitate of cupric sulfide
will form:
If the electrolysis is found to be complete, stop the rotation of

the anode but do not s'top the flow of current. Siphon the acid
electrolyte liquor from the cathode cup, but keep the dish filled
by the simultaneous addition of distilled water from a wash
bottle. About 500 cc. of water must usually be employed before
the ammeter will indicate that practically no current is flowing.
The current should not be stopped while washing the acid electrolyte liquor from the cathode, since the deposited copper is
slightly soluble in strong acids.
315
3. Remove the dish, wash again with distilled water by decantation,

followed by a little alcohol and ether, successively, dry for a few minutes
only, at from 80 to 100C., .and weigh.
After stopping the current, detach the cathode cup and wash it
thoroughly with water, alcohol, and ether successively, dry it in
an electric oven or. air bath for 2 or 3 min. at 80 to 100C., allow
the cathode to cool in a desiccator, and weigh it. The operations
of drying and weighing should be performed as quickly as possible, since the copper is subject to oxidation upon exposure to air.
After weighing the cathodc with the copper deposit, dissolve the
deposit completely by treating it with two successive portions
of about 10 cc. of concentrated nitric acid mixed with about
10 cc. of water using gentle heat to effect solution. Wash the
electrode successively with distilled water, alcohol, and ether
and dry again at 80 to 100C., allow it to cool in a desiccatbr,
and weigh it again. The difference between the weight of the
cathode and the weight of the cathode with the copper deposit
is equal to the weight of copper deposited. The cathode may
be weighed before the electrolysis is started and again after the
deposition of the copper is complete, but this procedure leads to
a slight inaccuracy since the anode dissolves slightly during the
electrolysis and the dissolved platinum is plated out on the
cathode.
Calculate the percentage Cu, CUS04, and CuS04.5H 20 to
which the sample assayed corresponds.
1. Explain how oxidation and reduction are involved in the assay of copper
sulfate.
2. Why is the electrolyte acidulated with sulfuric acid?
3. Why is the solution of electrolyte stirred?
4. Why should the current not be stopped until the acid electrolyte has
been washed out of the cathode dish?
5. How many coulombs of electricity would be required to completely
deposit the copper from 0.6 Gm. of pure CuS04.5H 20?
Exercise 93
Object.-Assay of Mercuric Chloride.

Materials Required.-An electrolytic apparatus with a mercury cathode
cup.
316
0.4 Gm. of mercuric chloride.

1 cc. of concentrated nitric acid.
1 cc. of hydrogen sulfide T.S.
10 cc. of toluene.
25 cc. of alcohol.
25 cc. of ether.
Procedure.-l. Dissolve from 0.3 to 0.4 Gm. of the salt, accurately
weighed, in 10 cc. of distilled water and transfer the solution to the cathode
cup, which has been previously weighed with its metallic mercury.
It is essential that the mercury cathode cup be thoroughly

cleaned. Before using, the cup, emptied of its mercury, should
be washed with concentrated nitric acid to dissolve the film of
amalgam which forms upon the platinum wire. The nitric acid
is removed by rinsing the cup successively with water, alcohol,
and ether and the cup is dried. The mercury cathode cup should
contain about 50 to 60 cc. of pure mercury. If the mercury
cathode is properly prepared, it may be used for a number of
determinations.
2. Dilute the liquid to about 20 cc. with distilled ws,ter, add 1 cc. of
nitric acid, which has been diluted with an equal volume of distilled water,
and 10 cc. of toluene, and pass through the solution a cUl:r5lnt of from 1 to
3 amperes and from 8 to 12 volts, stirring the s91ution by rotating the anode
from 500 to 600 revolutions per minute.
Mter weighing the mercury cathode cup accurately, atiach it to

the electrolytic apparatus, introduce the solution of mercuric
chloride into it, and wash the vessel in which the soIu.tion was
prepared with two 5 cc. portions of distilled water, adding each
of the wash solutions to that contained in the cathode cup. One
cubic centimeter of nitric acid is added to increase the conductivity of the electrolyte. The toluene is added to absorb the
liberated chlorine gas and prevent the latter from exerting a
solvent action on the platinum anode. After starting the rotation of the anode, turn on the current.
3. After 20 min., when the mercury is all removed from solution (to be
ascertained by removing a few drops of the solution and testing with hydrogen sulfide T.S.), wash with distilled water, with the aid of a siphon and
without interrupting the current, until the current drops to nearly zero.
Remove the cathode cup, wash the mercury with alcohol, then with ether,
remove most of the remaining ether with filter paper, dry over sulfuric acid,
and weigh. The increase in weight of the cathode CllP represents the
amount of mercury present in the quantity of the salt tak.en.
317
The method of testing the electrolyte for completion of the

electrolysis and the pro,cedure employed in washing out the
cathode cup have been described under the assay of copper
sulfate (page 313). The acid electrolyte liquor must be removed
before the current is stopped, since otherwise the acid will act
upon the mercury causing re-solution of some of the metal.
Calculate the per cent of mercury in and the purity of the
mercuric chloride assayed.
1. Explain the oxidation-reduction reactions which occur during the
electrolytic assay of mercuric chloride in terms of the ionic theory.

2. What substances other than toluene might be used to absorb the
chlorine liberated at the anode?
3. What assays of other official mercury salts may be performed electrolytically?
4. An unknown preparation of mercury weighing 0.75 Gm. when assayed
electrolytically yielded 0.2640 Gm. of mercury. To what per cent of each
of the following in the sample would the amount of mercury found correspond: HgCI, HgCh, and HgI 2 ?
Other Electrolytic Assays.-The general principles involved in

all assays by electrolytic methods are similar to those applied in
the two preceding exercises. Some of the following procedures
require explanation, however.
The electrolytic assay of the silver compounds offers no special
advantage over the gravimetric chloride precipitation method.
The silver is deposited from an electrolyte containing sodium
cyanide. In the case of silver nitrate, for example, the alkali
cyanide reacts with the silver salt to form a double cyanide:
AgNO s + NaCN--+AgCN + NaNOs

AgCN + NaCN--+AgNa(CNh
The double cyanide may be represented to ionize as follows:
AgNa(CN)2--+Ag+
+ Na+ + 2CN-
The preRence of the alkali cyanide increases the conductivity of

the electrolyte and, as has been found by experiment, causes the
formation of a firm adherent deposit of silver at the cathode
whereas in the absence of the cyanide a spongy, non-adherent
deposit is frequently obtained.
318
The mercury compounds insoluble in water for which electrolytic assays may be used, such as mercurous chloride, mercurous iodide, mercuric salicylate, and ammoniated mercury, are
electrolyzed from a solution of sodium sulfide, because mercury
sulfide is soluble in the presence of an excess of strong alkali
sulfide:
The complete deposition of mercury is determined by adding a

few drops of ammonium chloride to a drop of the electrolyte
contained in a depression of a spot plate. The ammonium
chloride reacts with the sodium sulfide forming ammonium
sulfide in which mercury sulfide is insoluble:
Mercury, if present in the electrolyte, is deposited as a black

precipitate. When the deposition of mercury from the cyanide
solution is complete, the mercury in the cathode cup is washed
with 2 to 3 per cent acetic acid solution to reI39ve the sodium
sulfide which is absorbed or dissolved in small amounts at the
contact surface between the mercury and electrolyte ~
Na 2S
+ 2CH2COOH~2CH2COONa + H 2S
The deposition of zinc from an electrolyte has been found to

take place best when a nickel cathode is employed and when the
deposition is effected from an electrolyte made alkaline with
sodium hydroxide. The reactions involved in the assay of zinc
chloride are typical of those which occur in all of the electrolytic
determinations of zinc. These reactions may be represented as
follows:
ZnCb + 2NaOH~Zn(OH)2 + 2NaCl
Zn(OHh + NaOH~Zn(OH)ONa + H 20
Zn(OH)2----+Zn++ + 20HThe rate at which the zinc is deposited is accelerated by heating
the solution of electrolyte, since zinc deposits rather slowly and
heat increases the speed of the ions.
319
TABLE XLIV.-SOME OFFICIAL SUBSTANCES WHICH MAY BE ASSAYED BY

Substance
U.S.P.
Copper sulfate ............
Copper sulfate, anhydrous
Mercury .................
Mercury, ammoniated .....
Mercury bichloride .......
Mercuric iodide, yellow ....
Mercuric oxide, yellow ....
Mercuric salicylate ........
Mercurous chloride, mild ..
Mercurous iodide, yellow ..
Mercury with chalk .......
Silver nitrate .............
Silver nitrate, toughened...
Zinc acetate ..............
Zinc chloride .............
Zinc oxide .. ,..............
.zinc sulfate ..............
Zinc sulfate, anhydrous ....
N.F.
Mercuric iodide, red .......
Mercuric oxide, red .......
Zinc phenolsulfonate ......
Amount
used,
grams
0.75 to 1.0
0.50toO.75
0.2 to 0.3
0.5 to 0.6
0.3 to 0.4
1.0 to 1.2
0.2 to 0.3
0.7 to 0.8
0.5 to 0.6
0.7 to 0.8
0.6 to 0.8
0.3 to 0.5
0.3 to 0.5
0.5
0.3
0.2
0.6
0.4
1.0 to 1.2
0.2 to 0.3
1.2
Amperage
used
Grams
equivalent
Voltage to 1 Gm.
used
of
deposited
metal
2.5
2.5
1.5 to
2 to
1 to
2 to
1.5 to
2 to
2 to
2 to
1.5 to
2"5
2.5
4 to
4 to
4 to
4 to
4 to
5 to
5 to
7 to
7 to
8 to
7 to
7 to
7 to
7 to
7 to
7 to
5 to
5 to
5 to
5 to
5 to
5 to
5 to
2
3
3
3
2
3
3
3
2
5
5
5
5
5
2 to 3
1.5 to 2
4 to 5
7
7
10
10
12
10
10
10
10
10
10
7
7
6
6
6
6
6
3.9281
2.5112
1.0000
1.2566
1.3535
2.2654
1.0798
1.1798
1.1768
1.6327
1.0000
1.5748
1.5748
3.3570
2.0849
1.2448
4.3988
2.4695
7 to 10
7 to 10
5 to 6
2.2654
1.0798
8.5011
'PART III
SPECIAL METHODS USED IN OFFICIAL
PHARMACEUTICAL ANALYSES
Quantitative analyses of crude drugs and of the products
derived from them are made to establish purity or to determine
the amount of therapeutically active constituents present for
the purpose of standardization. The special methods employed
in the analyses of this type may be classified as follows:
Chemical methods such as those employed in the determination
of ash, moisture, crude fiber, extractive obtained with different
solvents, the estimation of alkaloidal content, etc.
Physiological methods or those in which the effects upon
animals or animal tissues are measured and which are employed
in the absence of satisfactory chemical methods for standardization. The official drugs assayed by physiological methods are:
U.S.P. aconite, tincture of aconite, cod liver oil, non-qestearinated
cod liv:er oil, digitalis, digitalis powder, tincture of digitalis,
solution of epinephrine, solution of irradiated ergosterol, ergot,
fluidextract of ergot, extract of liver, solution of liver, purified
solution of liver, solution of parathyroid, solution of posterior
pituitary, stomach, and strophanthin and tests for the toxicity
and purity of arsphenamine, neoarsphenamine, and tryparsamide;
N.F. fluidextract of aconite, convallaria root, fluidextract of
convallaria root, ampuls of epinephrine hydrochloride, extract of
ergot, ampuls of posterior pituitary, fluidextract of squill, strophanthus, tincture of strophanthus, and suprarenal. Physiological methods require special apparatus for their performance
and a technique which is not chemical in nature; hence, a discussion of them does not fall within the scope of this text.
CHAPTER XX
ASH AND MOISTURE DETERMINATIONS

Ash Content.-Th~ ash content of a crude drug is generally
taken to be the residue remaining after incineration. It usually
represents the inorganic salts naturally occurring in the drug and
adhering to it, but it may also include inorganic matter added
for the purposes of adulteration. There is a considerable difference in the ash content of different drugs, but the difference varies
within narrow limits in the case of the same individual, hence an
ash determination furnishes a basis of judging the identity and
cleanliness of a drug and gives information relative to its adulteration with inorganic matter. Ash standards have been established
for a number of the official drugs; usually these standards set a
maximum limit on the total ash or on the acid-insoluble ash
permitted. The total ash is the residue remaining after incineration, while the acid-insoluble ash is that part of the total ash
which is insoluble in diluted hydrochloric acid. The Pharmacopoeia specifies that all vegetable drugs shall be separated by
mechanical means in so far as possible from lumps of dirt or other
foreign inorganic matter before they are ground or powdered.
When no ash standard is fixed for a given drug by the Pharmacopoeia or National Formulary, the amount of foreign inorganic
matter estimated as acid-insoluble ash must not exceed 2 per
cent of the weight of the drug.
Tpe ash or residue yielded by an organic chemical compound
is as a rule a measure of the amount of inorganic matter present as
impurity. In most cases, the inorganic matter is present in
small amounts which are difficult to remove in the purification
process and which are not objectiohable if only traces are present.
The careful control of temperature is the most important
analytical factor to regulate in making ash determinations. All
determinations should be made in such a manner as to duplicate
in so far as possible the conditions under which standards are
established. When an electric furnace is used for ignitions, the
323
324
following temperature equivalents which are only approximate

will be found of value: Very dull red heat = 500 to 550C.;
dull red heat = 550 to 700C.; bright red heat = 800 to 1000C.;
yellow red heat = 1000 to 1200C.; and white heat = 1200 to
1600C. Practically all of these temperatures are within the
limits of the ordinary Bunsen burner depending on the construction of the burner and the nature of the gas burned. When it
is considered that calcium carbonate is converted to calcium
oxide at about 825C., that calcium phosphate is converted to
calcium pyrophosphate at about 1550C., that lithium carbonate
forms lithium oxide at about 600C., and that potassium chloride
sublimes at about 1,500C., it becomes apparent that ignitions
should be carried out at low temperatures if possible.
Exercise 94
Object.-To Determine the Total and Acid-insoluble Ash

Content of Digitalis Leaf.
Materials Required.-2 to 4 Gm. of the ground drug.
2 ashless filter papers.
Procedure.-l. "Accurately weigh a quantity of the ground drug, representing from 2 to 4 Gm. of the air-dried material, in a -tared crucibie and
incinerate at a low temperature, not to exceed very dull redness, until free
I
from carbon, and determine the weight of the ash."
The crucible in which the drug is to be ignited should be heated

to dull redness until of constant weight, cooled in a de,siccator,
and weighed. This procedure drives off the moisture and the
absorbed gases which are present on the surface of the crucible.
The drug is incinerated at a temperature not to exceed a dull
redness. Very often, the ash consists largely of calcium carbonate formed from the calcium oxalate contained in the drug.
When the incineration is carried out at a dull red heat, the calcium
oxalate is converted into carbonate, but when the incineration
is carried out at a bright red heat, varying amounts of the
carbonate are converted into the oxide with consequent variable
results. If the ash contains alkali chlorides, which it frequently
does, there may be a loss of some chloride by volatilization when
a high ignition temperature is used.
2. "If a carbon-free ash cannot be obtained in this way, exhaust the
charred mass with hot distilled water, collect the insoluble residue on an
325
ashless filter paper, incinerate the residue and filter paper until the ash is
white or nearly so, then add the filtrate, evaporate it to dryness and heat the
whole to a low redness. If a carbon-free ash cannot be obtained in this way,
cool the crucible, add 15 cc. of alcohol, break up the ash with a glass rod,
burn off the alcohol, and again heat the whole to a low redness. Finally
determine the weight of the ash. Calculate the percentage of total ash from
the weight of the drug taken."
When the ignition residue is not heated above dull redness, it

is frequently very difficult to burn off all of the carbon, some of
which may become surrounded by fused inorganic matter thereby
keeping it from coming in contact with the oxygen of the air.
In such cases, the residue is treated with hot distilled water to
dissolve most of the inorganic matter, and the insoluble portion
is collected on an ashless filter and incinerated. The filtrate is
then added to this residue, evaporated, and the whole is ignited
to burn off any colloidal carbon which may have passed into the
filtrate. Occasionally, it may be necessary to treat the residual
mass with alcohol to disintegrate the fused lump, especially in the
ignition of resinous drugs.
The weight of the crucible and ash minus the weight of the
crucible gives the weight of total ash obtained from the sample
of drug taken. This total ash, which usually contains carbonates,
phosphates, sulfates, chlorides, oxides, etc., of calcium, magnesium, potassium, sodium, aluminum, iron, and other metallic
elements, does not necessarily represent all of the inorganic constituents of the drug, since ammonium salts, some alkali iodides
and nitrates, etc., are volatilized or converted into carbonates, etc.
3. "Boil the ash obtained with 25 cc. of diluted hydrochloric acid for
five minutes, collect the insoluble matter in a Gooch crucible or on an ashless
filter, wash with hot distilled water, ignite and weigh. Determine the per
c~nt of acid-insoluble ash calculated from the weight of drug taken."
The diluted hydrochloric acid dissolves the calcium carbonate,

alkali chlorideS', etc., leaving an acid-insoluble residue which
consists almost entirely of silica derived from the soil adhering
to the drug.
1. From what different sources may the ash of a drug be derived?
2. When no specific ash limit is set by the official standards, what per
c~nt of acid-insoluble ash is the drug permitted to contain?
326
3. Why should the incineration residue not be heated above a dull redness?
4. Name the important inorganic plant constituents which will not be
found in the ash.
6. What does the quantity of acid-insoluble ash in a drug indicate?
The ash content of chemicals is determined by ignition to

dull redness in the same manner as in the determination of the
ash content of a crude drug.
It is economically impracticable to make ash determinations
on large samples of expensive chemicals. However, it is necessary to control the amount of inorganic matter which may be
contained in such substances as alkaloidal salts, especially when
the substance is intended for hypodermic administration. For
this reason the official standards usually require that small
amounts of expensive substances be used and that the ash yielded
be negligible. The term negligible is defined as "a quantity
not exceeding 0.0005 Gm." A number of official substances,
with the quantity in Grams in parentheses, required to yield a
negligible amount of residue are: U.S.P. (0.1) atropine, (0.1)
atropine sulfate, (0.5) barbital, (0.5) carbromal, (0.5) cocaine,
(0.5) cocaine hydrochloride, (0.5) codeine, (0.5) codeine sulfate, (0.1) colchicine, (0.2) emetine hydrochloride, (0.1)
epinephrine, (0.2) ethylmorphine hydrochloride, (0.5), eucaine
hydrochloride, (0.1) homatropine hydrobromide, (0.5) morphine
sulfate" (0.2) pelletierine tannate, (0.5) phenobarbital, (0.1)
physostigmine salicylate, (0.1) pilocarpine nitrate, (0.5) procaine
hydrochloride, (0.1) scopolamine hydrobromide, (0.1) strophanthin, and (0.1) theophylline; N.F. (0.1) cotarnine chloride, (0.1)
hydrastine hydrochloride, (0.5) morphine hydrochloride, and
(0.1) pilocarpine hydrochloride.
The ash limits of most of the official vegetable drugs and
chemicals are indicated in Table XLV. The per cent of
ash given is the maximum permitted for each substance. Those
vegetable drugs fot which a maximum limit of 2 per cent of acidinsoluble ash is permitted are not included.
Residue limits are given in the official standards for a number
of soivents and preparations. The residue is determined by
evaporation and drying to constant weight at a specified temperature or by evaporation and ignition of the residue as indicated
in Table XLVI, page 329.
327

TABLE XLV.-OFFlCIAL SUBSTANCES WITH ASH LIMITS
Substance
U.S.P.
Acacia ................ .
Acetanilid .......... .
Acetophenetidin. . . .. . ..
Acid. acetylsalicyhc.. ..
Acid. acetyltannic..
..
Acid. benzoic .. .
Acid, citric .. .......... .
Acid lactic.. . .
. ..... .
Acid. oleic... ... . . . .. ..
Acid. salicylic .......... .
Acid. tannic ........... .
Acid. tartaric .......... .
Acid. trichloroacetic ... .
Agar ................. .
Albumin tannate.
Aloe .................. .
Aloin .............. .
Aminopyrine . .......... .
Ammonium benzoate .. . .
Ammonium bromide . ... .
Ammonium carbonate.
Ammonium chloride ... .
Ammonium salicylate ... ,
Antipyrine ............ .
Asafoetida .......... '"
Aspidium ............. .
Belladonna leaf. ...... .
Belladonna root ... .
Benzoin, Sumatra ..
Benzoin, Siam .... .
Betanaphthol. ... .
Caffeine ........... .
Caffeine, citrated . .. .
Camphor ......... .
Cannabis ....... . .
Capsicum ......... .
Cardamom seed.
Caraway ..... .
Charcoal. activated ... .
Chloral hydrate ........ .
Chlorobutanol. " ... .
Chrysarobin ........... .
Clove ............... .
Cotton. purified ........ .
Creosote carbonate ..... .
Dextrose ... .
Total
ash.
per
cent
>Acidinsoluble
ash,
per
cent
4.0
0.5
0.05
0.05
0.05
0.3
0.05
0.05
0.12
0.10
0.05
0.5
0.05
0.05
1.0
0.3
4.0
0.5
0.1
0.05
0.05
0.05
0.1
0.05
0.1
15.0
3.0
3.0
4.0
o 05
1.0
0.5
0.05
0.1
0.05
5.0
1.25
5.0
1.5
4.0
0.05
0.1
0.25
0.75
0.2
0.1
0.1
Total
Substance
U.S.P.
Digitalis ............. "
Elaterin ............... .
Ephedrine ............. .
Ephedrine hydrochloride.
Ephedrine sulfate ...... .
Ethyl aminobenzoate .. .
Glucose .............. "
Glycerin .............. .
Glycyrrhiza ............ .
Glycyrrhiza. extract of .. .
Guaiacol. ............ .
Hyoscyamus . .......... .
Iodoform .............. .
Iodine .............. .
Lactose ...... ....... .
Menthol. ........... .
Mercuric chloride 300C.
Mercuric oxide, yellow .. .
Mercuric salicylate . .... .
Mercurous chloride . .... .
Mercurous iodide, yellow
Mercury .............. .
Mercury, ammoniated.
Mercury succinimide . .. .
Methenamine .......... .
Methylthionine chloride.
Myristica ............. .
Myrrh ................ .
Paraffin, chlorinated .... .
Petrolatum ............ .
Petrolatum. white ...... .
Phenacaine hydrochloride
Phenol. ............... .
Phenol. liquefied. . . .. ..
Phenolphthalein ....... .
Phenolsulfonphthalein .. .
Phenyl salicylate ....... .
Pi~ tar ............... .
Prepared chalk ........ .
Pyrogallol. ............ .
Quinidine sulfate ....... .
Qninine ............... .
Quinine ethyl carbonate.
Quinine and urea hydrochloride ............. .
Quinine bisulfate ..
ash,
per
cent
Acidinsolu
ble
ash.
per
cent
5.0
0.1
0.1
0.1
0.1
o1
0.5
0.007
2.5
8.0
0.1
12.0
0.2
0.05
0.1
0.05
0.1
0.2
0.2
0.1
0.2
0.02
0.2
0.1
0.05
1.0
0.5
4.0
0.1
0.05
0.05
0.1
0.05
0.05
0.05
0.2
0.05
0.25
2.0
0.1
0.1
0.1
0.2
0.05
0.05
328
TABLE XLV.-OFFICIAL SUBSTANCES WITH ASH
Substance
Total
ash,
per
cent
U.S.P.
Quinine dihydrochloride.
Quinine suliate ......... .
Resin of podophyllum .. .
Resorcinol. ............ .
Resin ................. .
Saccharin, soluble ...... .
Santonin .............. .
Sarsaparilla, Mexican ... .
Senna ................. .
Serpentaria ............ .
Starch ................ .
Acidinsoluble
ash,
per
cent
0.05
0.05
1.5
0.05
0.05
0.5
0.1
4.0
3.0
10.0
0.5
Stramoni UIn .. . . . . . . . . . .
Strychnine sulfate ...... . 0.1

Sucrose ............... . 0.05
Sulfonethylmethane .... . 0.05
Sulfur, precipitated ..... . 0.3
Sulfur, sublimed ........ . 0.5
Sulfur, wl..shed ......... . 0.3
Terpin hydrate ......... . 0.05
Theophylline with ethylene diamine . ....... . 0.1
Thymol. .............. . 0.05
Thymol iodide ......... . 1.5
Valerian ..............
Vanillin ............... . 0.05
Veratrum viride ........
Wool fat .............. 0.1
l"I.1".
Acid, gallic ............ 0.1
Aletris ................ .
Animal charcoal, purified
4.0
Anise ................. .
Areca ................. .
Brucine sulfate ......... . 0.1
Buchu ................ . 1.0
Calamus .............. . 6.0.
Calumba .............. .
Camphor ,monobromated
0.05
Caramel. .............. . 8.0
Carmine .............. . 12.0
CauolphYIlum ......... .
Celery fruit ............ .
Chlorthymol. .......... . 0.05
Cimicifuga ............ .
Cinchouine sulfate ..... . 0.1
Cinchonidine sulfate .... . 0.1
Cinchophen ........... . 0.25
Coal tar .............. . 2.0
Colchicum corm .......
4.0
LIM1TS.-(Continued)
Substance
Total
ash,
per
cent
N.F.
Colocynth ............. .
Convallaria root ....... .
Coriander ............. .
Corpus luteum...... .... 6.0
Crocus.................
7.5
Cudbear. . . . . . . . .. .... 12.0
Damiana .............. .
Euonymus ............ .
Euphorbia ............. .
Fennel. ............... .
Gambir ............... .
Gamboge .............. .
0.1
Guaiacol carbonate......
Guarana .............. .
Humulus .............. .
Hydrastis ............. .
IchthammoI........ .... 0.5
Ipomoea .............. .
Iris .......... ......... .
Jalap ............ -..... .
10.0
4.0
0.5
5.0
3.0
3.0
1.0
0.5
6.0
due ................. .
1:::::
,.....
6.0
5.0
10.0
0.25
4.0
4.0
7.0
Pimenta .............. .
0.5
4.0
4.0
3.0
1.5
0.5
1.0
0.5
Pituitary, R"lterior ... , ..
4.0
1.0
Kamala .............. .
1.5
2.5
4.0
3.0
6.0
6.0
1.5
Kola .................. .
Leptandra .......... , .. .
Lobelia ............... .
Lupulin ............... .
Mastic ................ .
Matricaria . ........... .
Mullein leaves ......... .
Ovary and ovarian resi-
10.0
0.5
2.5
AcidinsolubIe
ash,
per
cent
7.0
Pituitary, whole ....... . 7.0
Plantago seed .......... . 4.0
Poplar bud ............ .
Quassia ............... .
Resin. ipomoea . ..... , ..
0.5
Rose ................. .
Salvia ................ . 10.0
Sassafras .............. .
Sassafras pith .......... .
Strychnine ............ . 0.1
Suifonemethane ........ . 0.05
Suprarenal. ........... . 7.0
Taraxacum ............ .
Thyme ..... " ......... .
Triticum ....... ' ...... .
Ulmus ................ .
Viburnum prunifolium .. .
0.4
1.0
1.0
0.5
1.0
1.0
5.0
0.5
4.0
4.0
3.0
1.0
3.0
329
TABLE XLVI.-SOME OFFICIAL SUBSTANCES WITH RESIDUE REQUIREMENTS
Substance
U.S.P.
Acetone ...... " ................ "
Acid, acetic .......................
Acid, diluted hydriodic .............
Acid, hydrochloric .................
Acid, nitric .......................
Acid, sulfuric .....................
Alcohol. .................... '" '"
Benzin, petroleum .................
Ether ............................
Ethyl oxide .......................
Water,
ammonia .......................
distilled ........................
distilled, sterilized ...............
orange flower ...................
rose, stronger ...................
N.F.
Antiseptic solution .................
Water ............................
Water, hamamelis .................
Water, redistilled ... , ..............
Amount
used, Gm.
or cc.
__Residue
dried at
c.
Official
requirement, Gm.
100
40
100
100
0.002
0.002
0.100
0.002
0.002
0.001
0.001
0.001
0.001
0.001
10
100
100
100
100
100
100
100
100
100
0.002
0.001
0.001
0.001
0.001
10
100
100
100
100
100
100
100
0.184
0.030
0.025
0.0005
50
20
5
10
30
2
40
50
50
50
100
100
Ignite
Ignite
Ignite
Ignite
Moisture Content.-The moisture content of crude drugs is

somewhat variable, since most vegetable drugs are more or less
hygroscopic. Some drugs take up as much as 5 per cent of their
air-dried weight when stored in a humid atmosphere. Generally,
assays are made to determine the proportion of active constituents contained in a drug in the condition in which it is purchased
or used. In accurate scientific work, however, and in those
cases where the drug is to be sold with a guaranteed assay, the
per cent of active constituent n'lust be calculated on the basis
of the moisture-free drug.
The method most commonly followed in determining the moisture content of a drug is to heat it at 100C. in an oven until
the weight becomes constant. Since, however, most vegetable
drugs contain variable amounts of volatile substances other than
330
water, such as essential oils, ethers, and esters, amines and

alkaloids in some instances, this method is not generally applicable except for rough estimations. When the drug contains
volatile matter other than water, the U.S.P. and N.F. directions
require that the volatile ether-soluble extractive shall be determined and the weight of the latter subtracted from the weight
lost by the drug upon drying, the difference to be taken as the
moisture content of the drug. Even this procedure is not free
from error. Since water is slightly miscible with ether, some pf
the moisture contained in the drug is extracted by the anhydrous
ether and determined as volatile ether-soluble extractive. A
better method, which was developed by the U. S. Forestry
Service, is commonly known as the xylene method, although
kerosene, toluene, and other organic solvents have been employed
in the plac~ of xylene. A modification of this method is official
as the moisture method by toluene distillation. It is the method
specified for the determination of the moisture content of thyroid
and is generally applicable to moisture determinations in glandular products and vegetable drugs containing 2 or more per cent
of moisture. This method has the disadvantage that a comparatively large amount of drug, in some cases 50,to 100 Gm.,
must be used for the determination in order to secure a v~lume
of water that can be measured conveniently without considerable
error.
Most of the official chemical compounds are relatively stable
in respect to their moisture content at room temperatures, i.e.,
they neither gain nor lose more than slight traces of water. Some
of them are hygroscopic and absorb water readily, e.g., aluminum
sulfate. Others lose a part or all of their water of crystallization
unless properly preserved, e.g., copper sulfate. Control of the
amount of water contained in chemicals is important in making
solutions of definite concehtration and in the determination of
the dosage of those substances used as medicaments.
The moisture content of chemicals is determined by drying
them to constant weight, at a specified temperature. In some
cases, the U.S,P. and N.F. specify the allowable per cent of
moisture in a chemical; when no limit is specified, 5 per cent of
moisture is permitted. For certain chemicals, a limit Qf tolerance
for loss of water of crystallization has been adopted.
331
Exercise 95
Object.-To Determine the Moisture Content of Acacia.

Materials Required.-l0 Gm. of Acacia.
Procedure.-l. Powder the acacia in a mortar.
In the case of other unground or unpowdered drugs, prepare

about 10 Gm. Df the official sample by cutting, or shredding, so
that the parts are about 3 mm. in thickness. Seeds or fruits
smaller than 3 mm. should be cracked. High-speed mills, since
they generate considerable heat, should not be used for preparing
the sample, and care should be taken that no appreciable amount
of moisture is lost during the preparation and that the portion
taken is representative of the official sample.
The sample should be ground in order to expose as large a
surface of the drug as possible so that the moisture may be driven
off readily.
2. Accurately weigh about 10 Gm. of the drug as prepared in a tared
evaporating dish. Dry at a temperature of 100C. for 5 hr., and weigh.
Continue the drying and weighing at 1 hr. intervals until the loss is not
more than 0.25 per cent in 1 hr.'s drying.
The moisture and any other volatile constituents of acacia are

driven off.
Calculate the per cent moisture contained in the sample of
acacia used.
1. Why is the loss in weight of a drug dried at 100C. not always a true
measure of its moisture content?
2. If an air-dried drug containing 10 per cent moisture yielded 2.5 per
cent ash calculated on the basis of the air-dried drug, what per cent ash
would the moisture-free drug con tain ?
Exercis~
96
Object.-Determination of the Moisture Content of Digitalis

by the Toluene Distillation Method.
Materials Required.-50 Gm. of digitalis leaf.
Toluene Moisture Apparatus.-Use a 500 cc. flask preferably of Pyrex
glass, a straight tube Liebig condenser of about 500 mm. length, and a
332
moisture-tube receiver calibrated to 0.1 cc. of the type illustrated (Fig. 59).
Clean the condenser and moisture tube with cleaning mixture (see page 5),
rinse with distilled water and then with alcohol, and dry them in an oven at
100C.
Procedure.-l. "Place in the flask an accurately weighed amount of the

drug to be tested, which it is estimated will yield from 2 to 4 cc. of water.
If the drug is likely to cause bumping, add enough dry sand to cover the
bottom of the flask. Add sufficient toluene to cover the drug
completely, usually about 75 cc., and connect the apparatus
as illustrated. Fill the receiving tube with toluene by pquring
it through the top of the condenser. Heat the toluene in the
flask until it boils, and distil slowly, about 2 drops per second,
until most of the water has passed over; then increase the
rate of distillation to about 4 drops per second."
Sufficient drug must be taken to yield from 2 to 4

cc. of water because smaller amounts would be
difficult to determine without a relatively large per
cent error. Thus an error of 0.05 cc. in the calibration or reading of the moisture tube would cause
an error of 5 per cent if only 1 cc. of water is
obtained and of 1.25 per cent if 4 cc. of water is
obtained. At the boiling point of t'oluene, 110 to
FIG. 59.- 111 ce., the moisture in the drug is volatilized.
Assembled The moisture distils with the toluene, ana as the
apparatus for
moisture de- vapors condense and drop into the moisture tube,
termination the water which is immiscible with and heavier than
by toluene
toluene separates and collects at the bottom of the
distillation.
moisture tube.
2. "When the water is apparently all over, wash down the condenser
by pouring toluene in at the top, continuing the distillation a short time to
ascertain whether any more water will distil, and if it does, repeat the washing of the condenser with toluene. If any water remains in the condenser,
remove it by brushing it down into the tube receiver with a tube brush
attached to a copper wire and saturated with toluene, at the same time
washing the condenser with toluene. Allow the receiving tube to stand
until cooled to room temperature and if any' drops of water still adhere to
the sides of the tube they can be forced down by a rubber band wrapped
around a copper wire. Finally read the volume of water and calculate to
determine the percentage which was present in the drug."
Practically all the moisture distils over during the first 4 or

5 min. of distillation. The condenser tube is washed down with
333

TABLE XLVII.--OFFICIAL SUBSTANCES WITH }4:0ISTURE LIMITS
Substance
U.S.P.
Acacia .............. .
Acid acetyltannic. . . . .
Acid. tannic.. . . . . . . . .
Acriflavine. . . . . . . . . . .
Acr i II a vi ne, hydrochloride ............
Agar ............... .
Albumin tannate......
Aloe ................ .
Alum. exsiccated. . . . . .
Barbital. soluble. . . . . .
C~ffeine. .. . .. . . . . . . . .
Caffeine. citrated. . . . . .
Caffeine with sodium
benzoate. . . . . . . . . . .
Calcium iodobehenate.
Calcium lactate... . . . .
Cantharides. . . . . . . . . .
C ....ein. R...... . . . . . .
Codeine..............
Codeine sulfate. . . . . . .
Dextrose. . . . . . . . . . . . .
Digitalis ............ .
Digitali powdered ... .
Emetine hydrochloride
Ephedrine hydrochloride ................
Ephedrine sulfate. . . ..
Ergot ............... .
Ethylhydrocupreine
hydrochloride ......
Ethylmorphine hydrochloride.. .. . . . . . . . .
Fluorescin. soluble. . . .
Gentian ............. .
Glucose ............. .
Histamine phosphate ..
Iodoform ............ .
Magnesium oxide .... .
Magnesium sulfate ...
Merbaphen .......... .
Methylthionine chloride ..............
Morphine sulfate ..... .
Phenacaine hydrochloride .............. .
Phenobarbital. ...... .
Temperature,
C.
Moisture
limit,
per cent
15
100
100
100
H2S0,
100
200
100
80
80
80
100
120
100
100
80
100
105
100
H2S0.
H2S0,
3
12
7
7
16
6
10
10
1
9
5
5
2
25 to 30
10
10
6
12
10
8
5
8 t<i 16
2
2
8
H2S0,
100
105
90
100
H2S0.
Ignite
Ignite
100
10
5
10
21
1
1
10
45 to 52
2
"
16
110
130
12
105
140
7
7
Tern ..
Substance
perature,
C.
Moisture
limit,
per cent
U.S.P.
Phenolsulfonphthalein.
110
1
Potassium carbonate . .
180
15
Potassium citrate . ... .
150
3 to 6
Potassium iodide. " .. .
100
1.5
Potassium and sodium
tartrate .... ....... .
150
21 to 26
Quinidine sulfate. '" ..
120
5
Quinine ....... , ..... .
100
15
Quinine bisulfate ..... .
24
100
Quinine dihydrochloride .............. .
100
3
Quinine ethyl carbonate ............... . H2S0,
2
Quinine sulfate ....... .
100
16.2
Scopolamine hydrobromide .............. .
100
13
SoaP ............... .
llO
36
Soap. powdered ...... .
10
llO
Soap, 8oft ........... .
52
llO
Sodium acetate ...... .
120
36 to 41
Sodium biphosphate .. .
10 to 15
100
Sodium carbonate. ..
monohydra ted ..... .
10 to 15
110
10 to 13
Sodium citrate ... " .. .
150
Sodium iodide ....... .
120
7
Sodium phosphate ... .
43 to 50
110
Sodium phosphate, exsiccated ........... .
110
5
Sodium stearate .... .. .
110
5
Sodium sulfate ....... .
51 to 57
120
Sodium thiosulfate ... .
32 to 37
100
Starch .............. .
14
Strychnine sulfate .... .
100
11.5
Theobromine with sodium salicylate .. .. .
110
10
Theophylline ........ .
100
9.5
Theophylline with
ethylene diamine ... . H,SO,
4.5
Thyroid ............. .
100
6
Tryparsamide ....... .
110 2.5 to 3.5
Wool fat ............ .
100
0.5
Wool fat. hydrous .... .
25 to 30
100
N.F.
Acid. gallic .......... .
12
100
Aluminum sulfate .... .
200
45 to 49
Ammonium hypophosphite .............. H 2SO,
3
334

TABLE XLVII.-OFFICIAL SUBSTANCES WITH MOISTURE LIMITS
(Continued)
Temper-
Substance
ature,
DC.
Moisture
limit,
Substance
per cent
Temperature,
DC.
Moisture
limit.
per Qent
- - - - - - - - -------11--------N.F.
Ammonium iodide .. ..
Ammonium valerate,
acid ............ .
Arecoline hydrobromide ............ .
Brucine sulfate ...... .
Calcium glycerophosphate ............. .
Calcium hypophosphite ............. .
Calcium phosphate ... .
Carmine ............ .
Charcoal. purified animal. .............. .
Cholesterol. R ....... .
Cinchonidine sulfate .. .
Cinchonine sulfate ... .
Cinchophen ......... .
Corpus luteum ....... .
Cotarnine chloride, .. .
Ethyl carhamate ...
Ferric glycerophosphate ......... .
Ferric hypo phosphite ..
Hydrastine hydrochloride .............. .
Lithium benzoate .... .
Lithium c,,,bonate ... .
Li thi um citrate .... .. .
Lithium slllicyiate .... .
Manganese glycerophosphate......... .
Manganese hypophosphite ............. .
Mercuric iodide, red .. .
R.
= reagent.
H2S0.
110
H2S0.
H2S0.
100
13
130
10
H,SO.
3
4
200
100
100
100
100
100
100
H,SO.
H,SO.
110
H,SO.
25
12
2
12
5
2
6
1
3
5
3
H,SO.
100
100
150
26
100
110
H2S0.
120
= dried
10
2
1
N.F.
Mercuric o)(ide. red. .. H2S0,
Methyl-rosaniline. . .
110
Morphine hydrochloride. . . . . . . . . . . . . . .
Ovarian residue . ..... .
Ovary .............. .
Papaverine hydrochloride ...............
Pilocarpine hydrochloride ...............
Pituitary. anterior ... .
Pituitary, whole .. .
Potassium chloride.
Potassium guaiacolsulfonate... ....
Potas::dum hypophosphite...
. ...... .
Potassium thiocyanate
Quinine hYdrobromide.
Quinine hYdrochloride.
Quinine salicylate . ... .
Resin of ipomoea . ..... .
Resin of jalap .......
Sabal. .............. .
Salicin.. .. . .
. .... .
Sodium ",senate ..... .
Sodium hYllOphosphite.
Sodium sulfate ....... .
Sodium thiocyanate . . .
Sparteine sulfate ..... .
Strontium bromide ... .
Strontium salicylate .. .
Sulfonmethane ....... .
Suprarenal . ......... .
Zinc iodide ........ .
100
7.5
15
6
6
H2S0.
H,SO.
100
6
6
1
H,SO.
H,S04
Ii
Ii
llO
100
llO
5
10
100
1100
i
4
100
10 to 16
H2S04
"150
3
3
3
.;
H,SO.
100
llO
100
200
22
32
3
6
5
over sulfuric acid.
toluene to remove water which may adhere to its inner wall.

If the condenser is cleaned as directed, little difficulty will be
experienced with water adhering to the wall of the tube. Some
drugs hold their moisture quite tenaciously. Consequently,
it is always well to continue distillation for 2 or 3 min. after
335
it is apparent from the test that practically all of the moisture

has distilled. Droplets adhering to the wall of a properly
cleaned moisture tube can be dislodged by rotating the tube
between the fingers or by tapping the tube gently on wood or some
other base. Volatile organic substances, such as volatile oils
and amines, .remain dissolved in the toluene and so are not
calculated as moisture. Since the result is read directly in
cubic centimeters, it is apparent that the per cent moisture would
be 2 X number of cubic centimeters of water when a 50 Gm.
sample is used.
1. .Consult Table XLVII and list five vegetable drugs, three glandular products and five chemical compounds the moisture contents of which
might be determined by the toluene distillation method.
2. Less than 1 part of water is soluble in 10,000 parts of toluene at 25C.
Would the water dissolved in the toluene when :qIoisture is determined in
a dmg by the toluene distillation method be appreciable if 100 ee. of toluene
is employed?
Certain other substances have definite limitations on the

amounts of volatile matter which they will lose when heated,
e.g., when ignited, the loss in weight of prepared calamine =
not more than 1 per cent, of kaolin = not more than 15 per cent,
of purified siliceous earth = not more than 10. per cent, of
purified talc = not more than 5 per cent, of lead monoxide =
not more than 4 per cent, and ichthammol dried to constant
weight at 100C. loses not more than 50 per cent of its weight.
The loss in weight in these cases cannot be considered as moisture
only, since adsorbed gases as well as other volatile matter are
driven off.
CHAPTER XXI
EXTRACTIVE AND CRUDE FIBER CONTENT
The amount of extractive which a drug yields to a given solvent

is often an approximate measure of the amount of a_ certain
constituent or group of related constituents which the drug
contains. In cases where this is the object sought, it is important
that a solvent be used which will not dissolve appreciable quantities of other substances than those sought in the extraction.
In some cases, the amount of a drug soluble in a given solvent is
an index of its purity; e.g., acacia must yield not more than 1 per
cent of water-insoluble residue to meet the official requirement.
For efficient and complete extraction, the solvent should be
divided into a number of relatively small portions and used
successively. Each portion should be brought into intimate
contact with all parts of the sample and thenJ:>e removed before
the addition of the next portion. The use of fresh portions of
the solvent which are not saturated with extractiv~ favors the
rapid and thorough removal of the solute with the minimum
quantity of solvent.
The complete removal of extractive matter from a drug by
this method, however, requires a large quantity of solvent in most
cases, and most volatile solvents are expensive. Consequently,
it is convenient and economical to be able to use the same portion
of solvent repeatedly. To do so necessitates the separation of
the solvent and solute after the extraction, the separated solvent
being employed in a second extraction. This is readily accomplished by means of a special form of extraction apparatus of
which the Soxhlet is a good example.
The Soxhlet (Fig. 60) is the apparatus generally used for
extraction with volatile solvents where small quantities of drug
are extracted as in quantitative work. The solvent is placed in
a weighed flask A, which is heated over an electric hot plate or
over a water bath in the absence of a gas flame when the solvent
is highly inflammable. The weighed sample of drug is placed
336
337
in a porous paper extraction thimble, which is then fitted into

the extraction tube B. A reflux condenser C, is attached to the
top of the extraction tube. The solvent vapors pass upward
from the flask A through the side tube D of the extractor to the
condenser. The condensate drips into the thimble
until a sufficient amount has accumulated to raise
the level of liquid in the extraction tube to the top
of the siphon tube E. The solution. which has
been standing in contact with the drug is then
discharged into the flask where the solvent is
vaporized again. The solvent passes through this
-cycle repeatedly, and the solute is collected in the ~
flask. The heat should be so adjusted that the
B
solvent passes around the cycle about once every
E
10 min. The top of the thimble shoulQ, be above
the level of the top of the siphon side tube. The
solvent must not be volatilized more rapidly than
it will filter through the porous thimble, or the
latter will overflow and carry insoluble matter over
mechanically. After the extraction is complete,
FIG.
60.Soxhlet extracthe flask is removed, the solvent is driven ,off until tion
apparatuB,
the flask and its contents are of constant weight,
and the percentage of extractive in the drug is calculated from
the weight of the residue.
In some cases, it is advantageous to deter'mine the insoluble
residue after extraction. The extraction thimble should be
dried and weighed accurately before placing the sample in it,
After the extraction is complete, it is dried with the residue and
weighed accurately again. Caution: Do not place thimbles
and residues saturated with ether or other inflammable or
explosive solvents in an electric oven until they have been
air-dried.
Volatile and Non-volatile Ether-soluble Extractive.-The
determination of the total 'ether-soluble volatile constituents
is applied to those drugs which contain volatile oils, while the
determination of the non-volatile ether-soluble constituents is
applied to those drugs the active constituents of which are associated with volatile matter. In either case, the determination is
only approximate.
338

Exercise 97
Object.-To Determine the Volatile and Non-volatile Ethersoluble Content of Clove.

Materials Required.-2 Gm. of clove.
50 to 100 cc. of absolute ether.
Soxhlet apparatus.
Procedure.-l. "Extract completely 2 Gm. of the prepared drug, dried
over sulfuric acid for not less than twelve hours, by subjecting it, during
twenty hours, to the action of absolute ether in a continuous extraction
apparatus,"
The drug is dried over sulfuric acid in a dessicator to remove

th! absorbed moisture. The extraction is continued for 20 hr.
to insure complete removal of the volatile oil and other ethersoluble constituents from the ground clove. Absolute ether
is used, since ordinary ether, which contains small, variable
quantities of water, 'dissolves some tannin, sugar, etc.
2. "Transfer the ethere::tl solution to a tared porcelain dish and ::tHow it
to evaporate spontaneously. Then dry it over sulfuric acid during eighteen
~
TABLEXLVIII.-OFFICIAL SUBSTANCES WITH ETHER-SOLUBLE ExTRACTIVE
Volatile, Non-volatile, To\al ether-soluble

per cent
per cent
extractive, per cent
Substance
U.S.P.
Capsicum ............ .
Cinnamon ..............
Clove ............... , ..
Ginger .................
Ginger, fluidextract of ...
Linseed .............. , ..
Myristica ......... , .....
12
.
.
.
.
.
.
2
15
4.5
4.5W/V
20
30
25
N.F.
Cacao, prepared .........

Coriander ........ , .....
Cubebs .................
Kamala ...............
LupuIin .. " .......... '"
Mastic .................
Salvia ............ , .....
Sandalwood, white .......
.
.
.
.
.
.
.
22
0.5
10
70
60
97
1.0
3.5
339
hours and weigh the total ether extract. Now heat the extract gradually
up to 110C. until the weight becomes constant; the loss in weight during
the heating represents the,volatile portion of the extract."
Since ether boils at 35C., it evaporates rapidly at ordinary

temperatures without carrying off more than negligible amounts
of the volatile extractive. The residue, dried for 18 hr. over.
sulfuric acid to get rid of the small amounts of water extracted
by the ether, represents the ether-soluble extractive of the clove.
This residue consists chiefly of volatile oil, eugenol, and resin.
When the residue is heated at llOC. until of constant weight,
the volatile oil and eugenol are volatilized, leaving resin, coloring
matter, etc., as the non-vol.atile ether-soluble extractive. The
difference in the weight of the total extractive and the nonvolatile residue is the volatile ether-soluble extract of the clove.
Alcohol-soluble Extractive.__:_Alcohol is a good solvent for
resinous matter. Consequently, the determination of the
alcohol-soluble extractive is most frequently employed to
determine approximately the amount of resin in those drugs in
which the resinous matter is the important constituent. In
some cases, the determination of the alcohol-soluble extractive
is considered officially as an assay process, e.g., asafoetida and I
kino, and in other cases, the extractive upon further purification
is employed in the assay, e.g., jalap.
Exercise 98
Object.-Assay of Benzoin.
Materials Required.-2 Gm. of benzoin.
100 cc. of alcohol.
Procedure.-l. "Place about 2 Gm. of Benzoin, accurately weighed, in a
tared extraction thimble and about 0.1 Gm. of sodium hydroxide in the
receiving flask of the extraction apparatus and extra-ct with -.alcohol in a
Soxhlet apparatus or other suitable extraction apparatus for five hours, or
until completely extracted."
Tlie alcohol dissolves the resins, benzoic acid, cinnamic acid,

styrene, benzaldehyde, vanillin, esters, etc., contained in the
balsamic resin. The NaOH in the receiving flask forms nonvolatile salts of the volatile acids.
2. "Dry the insoluble residue at 100 e. for thirty minutes and weigh.
Determine the amount of moisture in the drug by the toluene distillation
0
340
method, page 331, calculate the weight of moisture in the Benzoin and subtract this weight of moisture from the original weight of the Benzoin taken
for the assay. The difference between this result and the weight of the
residue determined above represents the alcohol-soluble extractive."
The residue should be dried and weighed in the extraction

thimble. When the extraction is complete, the thimble should
be removed from the apparatus and allowed to drain and dry in
air for about 5 min. to remove most of the alcohol. The residue
and thimble can be dried conveniently at 100C. in an electric
oven. Since moisture remaining in the marc as well as the alcohol
is volatilized at 100C., it is necessary to correct for the loss in
moisture by a moisture determination.
Calculate the per cent of alcohol-soluble extractive assuming
that the benzoin contained 3.20 per cent moisture.
The diluted alcohol-soluble extractive is determined in the
same way, using diluted alcohol as the solvent.
TABLE XLIX.-OFFICIAL SUBSTANCES WITH ALCOHOL-SOLUBLE EX'rRACTlVE
LIMITS
Substance
Alcohol used,
per cent
Extractive,
per cent
U.S.P.
Asafoetida ..............
Benzoin, Sumatra ........
Benzoin, Siam ...........
Bismuth and potassium
tartrate ...............
Bismuth subgallate .... '"
Kino ...................
Myrrh ..................
Rhubarb ................
94
94
94
50
75
90,
94
94
94
94
49
(n.m.t.) 0.5
(n.m.t.) 0.5
73
94
94
94
94
95
95
49
25
65
60
85
75
80
40
12
60
30
30
N.F.
Chionanthus .. ' ..........

Gamboge ...............
Gambir .................
Guaiac .................
Manna .................
Mastic ............... '"
Poplar bud ..............
Vanilla .................
(n.m.t.) = not more than.
341
Water-soluble Extractive and Water-insoluble Residue.The determination of the water-soluble extractive content is
applied to those drugs one or more of the most important constituents of which are soluble in water, e.g., aloe, gambir, anet
kino. The determination is performed in the same way as the
determination '~f the alcohol-soluble extractive. The determination of the amount of water-insoluble residue is used as a
test for the purity of some substances. Thus, it is required
that acacia yield not more than 1 per cent, aloin yield not more
than 1.5 per cent, acetyltannic acid yield not more than 6 per
cent, and acriflavine yield not more than 0.5 per cent of waterinsoluble residue.
Exercise 99
Object.-Assay of Aloe.
Materials Required.-2 Gm. of aloe.
Procedure.-Proceed as directed in the assay of benzoin, Exercise 97,
using distilled water instead of alcohol as the solvent.
Barbaloin, isobarbaloin, and other active constituents of a

glycosidal nature dissolve in the water, leaving resinous matter,
etc., undissolved.
The U .S.P. requires that aloe contain not less than 50 per cent
of water-soluble extractive. Calculate the percentage of watersoluble extractive contained in the sample examined and compare
the result with the official requirement.
TABLE L.-OFFICIAL SUBSTANCES WITH WATER-SOLUBLE EXTRACTIVE LIMITS
Water-soluble
Extractive,
Per Cent
Substance
U.S.P.
Aloe .........................................., ....
Gentian ..........................................
Ginger ...........................................
Kino .............................................
50
30
12
80
N.F.
Gambir ........................................... 70
Purified Petroleum Benzin Extractive.-Purified petroleum

benzin is a good solvent for fats and fatty oils. The determination of the amount of extractive with this solvent is performed
as follows:
342
"Extract completely about 2 Gm. of the prepared drug,

accurately weighed, by subjecting it during twenty hours to the
action of purified petroleum benzin in a continuous extraction
apparatus. Transfer the benzin solution to a tared porcelain
dish and allow it to evaporate spontaneously. Then dry it
over sulfuric acid for eighteen hours and weigh. Calculate the
percentage of anhydrous extractive from the weight of drug
taken."
Colocynth is the only official drug for which a petroleum
benzin extractive limit is given. In this case the determination
serves as a check on the amount of seeds contained in the
colocynth pulp, since the seeds contain a large amount of fatty
matter.
Crude Fiber.-The crude fiber content of a drug is the residue,
consisting chiefly of cellulose, which remains undissolved after
successive treatment with boiling acid and alkali. The determination of crude fiber is of considerable importance in the
examination of certain drugs and particularly of spices, since the
commonly used adulterants consist of waste or refuse material
derived from the drugs or spices themselves or from other food
products. Frequently, this material is the outer cellular layer
or protective coating which contains a larger pI\Oportion of
lignified tissues and, consequently, more crude fiber than the
part official.
Exercise 100
Object.-To Determine the Crude Fiber Content of Clove.

Materials Required.-2 Gm. of clove in fine powder.
200 cc. of 1.25 per cent H 2SO .
200 cc. of 1.25 per cent NaOH.
Procedure.-l. "Exhaust a weighed quantity of the prepared drug
representing about 2 Gm. of the drug, with ether or use the residue from the
determination of the volatile ether-soluble extractive."
The drug should be powdered as finely as possible so that the

acid and alkali will penetrate the innermost tissues and thus
insure complete removal of soluble material. It is exhausted
first with ether to remove fats and waxes, which, being immiscible
with aqueous solutions, tend to prevent the penetration of the
acid and alkali solution into the drug particles.
EXTRACTIVE AND CRUDE FIBER CONTENT-
343
2. "Add 200 cc. of boiling sulfuric acid solution, adjusted to exactly

1.25 per cent by titration, to the ether-exhausted drug, in a 500 cc. flask,
and connect the flask with a reflux condenser, the tube of which passes only
a short distance below the rubber stopper, into the flask. Heat the mixture
to boiling and continue the boiling exactly thirty minutes. Then filter
the mixture through a linen or hardened-paper filter and wash the residue
on the Mter with. boiling distilled water until the washings are no longer
acid."
By diluting exactly 51 cc. of normal sulfuric acid or an equivalent amount of a standard acid of known normality to 200 cc.
at 25C., 200 cc. of 1.25 per cent sulfuric acid may conveniently
be prepared. The prepared acid solution should be measured
and heated to boiling before it is added to the drug. When
the acid solution is added to the powdered, ether-exhausted
drug contained in a cold flask, the solution is cooled below its
boiling point. The mixture should be heated rapidly to bring
it to the boiling point again, and thereafter the flame should be
cut down to the minimum required to maintain slow, steady
boiling. Since the amount of material dissolved by the acid
solution is partly dependent upon the length of time that the
latter is heated in contact with the sample, it is important that
the mixture should be boiled exactly 30 min., the time period
being taken from the time the mixture starts to boil and not
from the time the hot acid solution is added. The flask should
be rotated gently from time to time while the mixture is being
boiled to bring particles of the drug which adhere to the walls
of the flask into contact with the acid. A current of air, passed
into and beneath the surface of the mixture, through a capillary
tube, will frequently prevent excessive frothing. The acidinsoluble residue is collected on a filter and washed until free of
acid, since the latter, if present, would neutralize~some of and,
consequently, reduce the concentration of the alkali used in the
next step of the procedure.
3. "Rinse the residue back into"the flask with 200 cc. of boiling sodium
hydroxide solution, adjusted to exactly 1.25 per cent by titration and free
from sodium carbonate. Again heat the mixture to boiling and continue
the boiling exactly 30 min. under the reflux condenser as described under
the treatment with acid, then rapidly filter through a tared filter, wash the
residue with boiling distilled water until the last washing is neutral, dry it at
110C. until of constant weight, and note the weight."
344
The alkali solution may be prepared conveniently by diluting

70 cc. of recently standardized, carbonate-free, normal sodium
hydroxide or an equivalent quantity of standard alkali solution
of known normality to exactly 200 cc. The residue from the
acid treatment in step 2 should be boiled with alkali in a manner
duplicating as nearly as possible that used in the boiling with acid.
The residue is then collected on a filter, preferably in a Gooch
crucible, washed free of alkali, dried to drive off moisture, and
weighed accurately. The dried residue consists chiefly of cellulose and small amounts of hemic ell uloses, pentoses, inorganic
matter, etc.
4. Incinerate the dried residue, and weigh the ash.
The fibrous matter should be incinerated cautiously at first and.

then at a low red heat until the carbon is completely burned off
as indicated by the formation of a white ash. The cellulose, etc.,
is burned when the fiber is incinerated, leaving the inorganic
matter. The weight of the ash obtained subtracted from the
weight of the residue found in step 3 gives the weight of the crude
fiber.
The Pharmacopoeia requires that clove contain not more than
10 per cent of crude fiber. Calculate the perce~tage of crude
fiber contained in the sample examined and compare the results
with the official requirements.
Salvia is required to yield not more than 25 per- cent of crude
fiber. The ether-insoluble residue of prepared cacao of the
National Formulary is required to contain not more than 7 per
cent of crude fiber.
1. What is meant by the term crude fiber?
2. Why should the sample used be finely ground and exhausted with
ether before it is boiled with acid and alkali?
3. Why should the boiling with 1.25 per cent acid and alkali be continued
for exactly 30 min. ?
4. What substances remain undissolved by the acid and alkali?
CHAPTER XXII
CONSTANTS OF FATS, FATTY OILS, WAXES, BALSAMS,

RESINS, ETC.
The methods of analysis of fatty substances, waxes, resins, etc.,
usually consist of the determination of a number of physical
and chemical properties or values commonly known as constants,
although they are constant only within certain limits. These
constants, when taken in conjunction with the color, odor, taste,
and special identity tests for the given substance and for common
adulterants, are the basis upon which the purity and quality of
these substances are judged. Although the following determinations are not required of all of the official substances which fall
under the above caption, they are generally used in the examination of such substances.
The specific gravity, solubility, melting point, congealing point,
refractive index, and optical activity determinations are performed according to the methods which have been discussed
previously (pages 203 to 256).
Preparation of the Sample of Fats and Oils.-" If a sample
of oil shows turbidity due to separated stearin, warm the container in a bath of water at pOC. until the turbidity has disappeared ~nd the oil is clear. Thoroughly mix the clarified
oil before weighing the samples. If the turbidity is due to other
than separated stearin, i.e., if the oil does not become clear on
warming, filter it thr~>ugh dry filter paper in a funnel contained
in a hot water jacket. Weigh out at one time as many portions
as are needed for the various determinations, using preferably a
bottle having a pipette dropper, or a weighing burette. Keep
the sample molten, if solid at room temperature."
Acid Value.-The acid value, also known as the acid number
and as the acidity index, is defined as the number of milligrams
of potassium hydroxide necess.ary to neutralize the free acids in
1 Gm. of oil, fat, wax, resin, balsam, or similar organic substance
345
346
of complex composition. In other words, it gives the amount of

potassium hydroxide, expressed in tenths of 1 per cent, required
to neutralize the free acids in a substance. The acidity may also
be expressed as the number of cubic centimeters of 0.1 N NaOH
required to neutralize the free acid in 10 Gm. of substance. This
value is determined by titrating a weighed sample of the substance, contained in an alcoholic or in an alcohol-ether solution,
with standard alkali solution, using phenolphthalein as the
indicator. For the titration, 0.5 N, 0.1 N, or 0.02 N alkali may
be employed, but a solution of 0.1 N concentration is most
suitable in the majority of the official determinations. In the
case of substances like the balsams and resins, a mixed solvent
such as alcohol and ether may be used advantageously, since the
coloring matter which interferes with the observation of the end
point is dissolved in the ethereal layer when the alkali is added.
Solid fats and waxes are usually melted on a water bath and
titrated while hot.
The presence of free acids in the oils, fats, and waxes is due
chiefly to the hydrolysis of the esters composing them and
is caused by chemical treatment, by bact~ial action, or by
the catalytic action of light and heat. As a rule, fresh or recently
prepared fatty substances contain little or no free ~cids. Upon
aging, the acid value increases slowly at first and more rapidly
later, especially if the substance is not well protected from
the simultaneous action of light and air. The acid values
of the official fatty substances are quite variable; consequently,
the official standards fix ma~mum limits, in most cases, which
if exceeded indicate that the substances have undergone hydrolytic decomposition in their preparation, purification, or during
the period of storage. High acid values are not necessarily an
indication of rancidity, since the latter is a result of the action of
the air, or possibly bacteria, on the liberated fatty acids.
A minimum acid value or a minimum and.a maximum acid
value is generally given in the official standards for substances
which are balsamic or resinous in character. The value of these
substances is usually based, in part at least, upon their content
of free acids; e.g., the "Rcid value of tolu is required to be not
less than 112 and not more than 168. Since a good grade of
tolu usually contains from 12 to' 15 per cent of cinnamic and
CONSTANTS OF FATS, FATTY OILS, WAXES, ETC.
347
benzoic acid in the free state, an acid value of less than 112 would
indicate that the acid content of the balsam was low and thll.t it
was of inferior quality or adulterated. On the other hand, an
acid value greater than 168 would indicate adulteration with
some substance having high acid value, such as certain resins.
Exercise 101
Object.-To Determine the Acid Value of Rosin.

Materials Required.-About 1 Gm. of rosin.
0.5 N potassium hydroxide.
Procedure.-l. Pulverize the rosin in a mortar and dissolve about 1 Gm.
of the powder, accurately weighed in from 40 to 50 cc. of neutral alcohol.
The rosin is powdered to facilitate its solution. Commercial

alcohol is frequently acid in reaction and it should always be
tested before the sample is dissolved. This test may convenientlybe carried out in this case by adding 1 cc. of phenolphthalein
indicator solution to the alcohol before the rosin is dissolved.
If the alcohol is found to be acidic, it should be rendered neutral
to phenolphthalein by the addition of the standard alkali
solution.
In the titration of substances which form colored solutions,
the end point may frequently be made observable by dilution
with alcohol to a volume of from 100 to 200 cc.
2. Add 1 cc. of phenolphthalein indicator solution (if it has not already
been added in testing the neutrality of the alcohol) and titrate the solution
with 0.1 N NaOH, agitating the mixture continuously.
In the titration of fatty substances, it is necessary to shake the

mixture thoroughly after the addition of each portion of alkali to
secure complete extraction of the fatty acids from the immiscible
oily layer. This precaution is not of great importance in the
titration of rosin, however, since rosin is soluble in alcohol and
also in alkalies.
The alkali reacts with the dibasic abietic acid formed from the
abietic acid anhydride and with the other acids contained
in the rosin with the formation of so-called rosin soaps and water.
It has been suggested that the acid value of rosin can be more
accurately determined in alcoholic solution, using alcoholic
348
QUANTITATIVE PHARMA.CEUTICAL CHEMISTRY
K6H in the titration, since the presence of water in the above

determination causes dissociation of the rosin soap with consequent low results.
The calculation of the acid value is simple; e.g., if a 1 Gm.
sample of rosin required 30 cc. of 0.1 N NaOH for titration, the
5 612
acid value would be 30 X1 .
= 168.36, where 5.6] 2 is the
number of milligrams of KOH equivalent to 1 cc. of 0.1 N NaOH
solution.
1. Define the term acid value.
2. Why must the alcohol or ether used as a solvent for fats, resins, etc.,
in acid value determinations be neutral?
3. Look up the U.S.P. test for free fatty acids under h"trd and explain
why this test may serve as a criterion of the purity of this fat.
4. Why do the official standards set up maximum acid values for most of
the official fats and oils and minimum values for most of the resinous
substances?
TABLE LI.-AcID VALUlil LIMITS OF OFFICIAL SUBSTANCES
Substance
Amount
used,
Gm.
Alkali
used
Official
rtquircment,
cid value
U.S.P.
Acid, oleic ................
Copaiba ..................
Peruvian balsam ..........
Rosin ...................
Soap, hard (acids) .........
Soap, soft (acids) ..........
Storax, American ..........
Storax, Levant ............
Tolu balsam ........ I .
10
2
1
10
10
10
1
1
1
White wax ................
Yellow wax ...............
N.F.
Mastic ...................
Resin of ipomoea ..........
10
2
0.1 N NaOH
0.5NKOH
0.5 N NaOH
0.1 N NaOH
0.1 N NaOH
0.1 N NaOH
0.5 N NaOH
0.5 N NaOH
0.5NKOH
(alcoholic)
.0.5 NKOH
(alcoholic)
0.5NKOH
(alcoholic)
188 to 200
28 to 95
56 to 84
Not less than 150
185 to 205
190 to 205
38 to 85
56 to 85
112 to 168
0.1 N NaOH
0.5 NKOH
Not less than 50

8.5 to 18
17 to 23
18 to 24
349
o. If a 2 Gm. sample of cod liver oil required 4.5 cc. of 0.02 N NaOH in
the titration of the free fatty acids, would the oil conform to the official
purity requirement? What would be the acid number of the oil?
6. Calculate the minimum and maximum acid values permitted under
the tests for purity of chaulmoogra oil.
In addition to the listed substances for which definite acid
value limits are given in the official standards, there are a number
of substances for which the maximum content of free fatty acids
is fixed by the volume of the standard alkali solution required
in their titration; e.g., 2 Gm. of cod liver oil must require not
more than 1 ee. of 0.1 N NaOH, and 1 Gm. of prepared suet
should not require more than 0.6 cc. of 0.1 N NaOH to neutralize
the free fatty acids.
Saponification Value.-The saponification value, saponification
number, or Koettsdorfer number, as it is sometimes called from
the originator of the process, is defined as the number of mt'lligrams
of potassium hydroxide required to neutralize the free acids and
saponify the esters contained in 1 Gm. of oil, fat, wax, or other
substance of similar composition. This value represents the
amount of potassium hydroxide, expressed in tenths of 1 per cent,
required to neutralize the total free and combined acids in 1 Gm.
of the substance, or, in other words, it is ten times the percentage
of potassium hydroxide required to neutralize all of the acids
contained in the sample afte.r saponification. Since the natural
fats and oils consist of mixtures of glyceryl esters of the higher
acids, their saponification values do not differ greatly. The
determination of the saponification value, however, serves to aid
in the detection or" the presence of the glycerides of acids containing less than 16 or more than 18 carbon atoms, since the value
of this constant is inversely proportional to the mean molecular
weights of the acids present. In some. cases, it may a,_lso indicate
adulteration of the sample with ul,lsaponifiable matter, such as
mineral oil.
Exercise 102
Object.-To Determine the S'aponification Value of Cottonseed

Oil.
Materials Required.-2 Gm. of cottonseed oil.
50 cc. of 0.5 N alcoholic potassium hydroxide (see page 365).
About 50 cc. 0.5 N hydrochloric acid.
350
"If the oil has been saturated with carbon dioxide for the purpose of preservation, it should be exposed in a shallow dish in a vacuum desiccator for
twenty-four hours before the portions are weighed for this determination."
Procedure.-l. Place from 1.5 to 2 Gm. of the sample, accurately weighed,
in a flask of from 200 to 250 cc. capacity, and add to it exactly 25 cc. of
alcoholic 0.5 N potassium hydroxide. rnsert into the neck of the flask, by
means of a perforated stopper, a glass tube from 70 to 80 cm. in length and
from 5 to 8 mm. in diameter, and heat the flask on a water bath for 72' hr.,
frequently rotating the contents.
If the sample is clear, it need not be filtered as a rule. When

the presence of extraneous matter, however, makes it necessary
to filter the oil, the operation may be carried out easily by means
of a suction filter. The sample of oil should be introduced into
the weighed flask by means of a dropper to avoid the possibility
of a portion adhering to the neck of the flask, and the whole
reweighed. The long glass tube serves as an air condenser to
prevent the escape of alcohol vapor. An ordinary waterjacketed condenser may be used with equally satisfactory results.
The mixture of oil and 0.5 N alcoholic KOH is heated on a water
bath and mixed by imparting a rotatory motion to the flask
from time to time in order to accelerate the- saponification.
The alcoholic KOH solution used should not be deeply colored
(see page 366), since the color would interfere wi~h tHe observation of the end point in the titration of the saponification mixture.
Alcoholic KOH, rather than an aqueous solution, is employed,
because the oils are more soluble in alcohol than in water, and
because the products of saponification are completely soluble in
alcohol, whereas when aqueous solutions are used, the unsaponifiable matter remains insoluble.
When saponification is complete, the mixture should be clear
and free from oil globules. Although the glyceryl esters present
in fatty oils vary grea:tly with respect to their acid components,
the character of the saponification reaction may be illustrated
as follows:
CSH6(C17HssC02)S
glyceryl oleate
+ 3KOH~CsH6(OH)s + 3C 17H 33C02K

glycerin
potassium oleate
2. Then add 1 cc. of phenolphthalein T.S. and titrate the excess of

potassium hydroxide with 0.5 N hydrochloric acid.
351
The approximate amount of alkali consumed in the saponification of the oil is found by subtracting the volume of 0.5 N ReI
used in the residual titration from the volume of 0.5 N alcoholic
KOR originally added. Sulfuric acid should not be substituted
for hydrochloric acid, since potassium sulfate would be precipitated from the alcoholic solution and this would interfere with
the observation of the end point.
3. Make a blank test at the same time, using exactly the same amount
of alcoholic 0.5 N potassium hydroxide. The difference in the number o~
cubic centimeters of 0.5 N hydrochloric acid consumed in the actual test
and the blank, multiplied by 28.06 and divided by the weight of the sample
taken, gives the saponification value.
The blank,test should be carried out at the same time as the

sample is run, using similar flasks, boiling for the same length of
time and under similar conditions, except that the oil should be
omitted. This is done to eliminate as far as possible errors from
every source such as those which would be introduced by the
absorption of CO 2 by the alkali or by the alkalinity of the glass.
The saponification value may be calculated as in the following
example: 1.532 Gm. of cottonseed oil saponified with 25 cc. of
0.5 N alcoholic KOR required 11.0 cc. of 0.5 N RCI to hack
titrate the excess alkali. In the blank test 21.5 ce. of 0.5 N RCI
were required to titrate the alkali. Therefore a quantity of
.
h y d roXlde correspondmg to (21.5 - 11.0)
0.0561 X
potaSSIUm
2
1,000 = 294.5 mg. of KOR was consumed in the saponification,
the number of milligrams of KOR required to saponify; 1
Gm. of the oil is 294.5/1.532 = 192.2 mg. of KOR. The saponification value of the sample of cottonseed oil is therefore 192.2.
Since 1 cc. of 0.5 N HCI is equivalent to 1 ce. of 0.5 N KOH
~nd
or to 2
~6i~~00 =
0.02806 Gm.
~OR,
the saponification value
may be calculated more simply from the formula:

ce. 0.5 N HCI X 28.06
.fi.
I
10.5" X 28.06
wt. of sample
= sapom catlOn va ue, or
1.532
= 192.2 mg. KOH required to saponify the esters and neutralize the free fatty acids in 1 Gm. of the sample.
352
The saponification values of the official fats, waxes, resins, etc.,

are deter:rpined in the same way.
1. Define the term saponification value.
2. Why should a blank determination be run parallel with the determination of the saponification value of tho sample?
3. Consult the following table and enumerate several oils which, if used to
adulterate croton oil, might lower its saponification value.
4. A sample of chaulmoogra oil weighing 1.600 Gm. saponified with 25 cc.
of 0.4 N KOH required 9.0 cc. of 0.5 N HCl to titrate the- excess KOH.
In the blank determination 20 cc. of 0.5 N HCI was required to titrate the
alkali. Calculate the saponification value of the sample. Does the value
found correspond with the U.S.P. requirement?
6. Need the alcoholic KOH used in a saponification value determination
be of an exact known normality?
6. How is the saponification value of balsam of Peru determined in the
U.S.P.? Does the method correspond with that given in the preceding
exercise?
.
TABLE LII.-SAPONIFICATION VALUE LIMITS OF OFFICIAL SUBSTANCES
Saponification
value
Amount
us~sl,
Substance
Gm.
o
U.S.P.
Balsam of Peru ................
Castor oiL ....................
Chaulmoogra oil. .......... ...
Cod liver oil. ..................
Corn oil. .....................
Cottonseed oil. ................
Ethyl chaulmoograte ...........
Expressed oil of almond .........
Lard .........................
Linseed oil. ......... , ........
Oil of thedbroma ...............
Olive oil. ............
Prepared suet .................
Storax ........................
Tolu balsam ...................
N.F.
Croton oil ................... ~ .
Resin of ipomoea ..............
Sesame oil. ...................
o
3
1.5 to 2
1.5 to 2
1.5 to 2
1.5to2
1.5to2
1.5to2
1.5 to 2
1.5to2
1.5to2
1.5to2
1.5to2
1.5to2
1.5 to 2
1
235 to
179 to
196 to
180 to
188 to
190 to
190 to
191 to
195 to
187 to
188 to
190 to
193 to
160 to
154 to
1.5 to 2
1.5to2
1
200 to 215
170 to 190
188 to 193
238
185
213
192
193
198
196
200
203
195
195
195
200
200
220
353
Ester Number.-The ester number, or ester value, is defined

as the number of milligrams of potassium hydroxide required to
saponify the esters in,l Gm. of a fat, oil, wax, balsam, resin, or
aimilar organic substance. In those substances which do not
contain free acids, the ester number is equal to the saponification
value. When. free acids are present, however, the ester value
is given by the difference between the acid and saponification
values.
The ester value is of particular importance in the analysis
of beeswax, since it serves, in many cases, to indicate the presence
of adulterants such as paraffin. Considered in conjunction with
the acid value, the ester value may aid in the detection of such
adulterants as rosin and stearic acid in the wax.
The United States Pharmacopoeia gives the following directions for the determination of the ester value:
II Shake from l.5 to 2 Gm. of the substance, accurately weighed
in a 200 to 250 cc. tared flask, with from 20 to 30 cc. of alcohol,
add 1 cc. of phenolphthalein T.S. and titrate with half-normal
alcoholic potassium hydroxide until the free acid is neutralized.
Add exactly 25 cc. of half-normal alcoholic potassium hydroxide
and proceed as directed under" Saponification Value" beginning
with "Insert inthe neck of the flask" and omitting the further
addition of phenolphthalein T.S. The difference in the number
of cc. of half-normal hydrochloric acid consumed in the actual
test and in the blank, mUltiplied by 28.06 and divided by the
weight of the substance taken, gives the ester value."
1. Define the term ester number.
2. If a sample of beeswax is found to have an acid number of 20.4 and a
saponification value of 89.8, what would be the es~er number of the sample?
8. If a sample of white beeswax has an acid value of 18.45 and an ester
value of 74, what would be the saponification value of the sample?
TABLE LIII.-OFFICIAL SUBSTANCES WITH THEIR REQUIRED ESTEIr.
'NUMBERS
Official Ester Value

Substance
U.S.P.
White wax. . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . .. 72 to 79
YeHow wax. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 72 to 77
N.F.
Resin of ipomoea ........................... 160 to 180
354
Un saponifiable Matter.-The term unsaponifiable matter is

defined in the Pharmacopoeia as the substances present in oils
or fats that are not saponified by alkali hydroxides and are insoluble
in water.
When oils and fats are saponified, there remains a small amount
of residue which may consist of phytosterol in the vegetable oils
and fats, of cholesterol in the animal oils and fats, and in some
cases, of unsaponifiable substances added for the purpose of
adulteration. The normal unsaponifiable matter present in
most oils and fats dissolves in the alcoholic soap solution obtained
upon saponification. When a large amount of unsaponifiable
matter is present, however, it may separate out and float on the
surface of the mixture.
The determination of the amount of unsaponifiable matter
present in certain oils such as linseed oil and cod liver oil may be
indicative of the quality and purity of the oil. Thus, the addition
of petroleum oils, a common adulterant, to linseed oil would
greatly increase the unsaponifiable matter content of the latter.
The Pharmacopoeia gives the following procedure for the
determination of the amount of unsaponifiable J?}atter in a fat
or wax:
Weigh 5 Gm. of the oil or fat into a 250 cc. Erlenmey;er flask,
add a solution of 2 Gm. of potassium hydroxide in 40 cc. of
alcohol, and heat under a reflux condenser for 2 hr. keeping the
alcohol gently boiling. Evaporate the alcohol on a water bath,
dissolve the residue in 50 cc. of hot distilled water, and transfer
the solution to a separatory funnel, rinsing the flask with two
25 cc. portions of hot distilled water which are added to the
separator. Cool to room temperature, and extract with two
successive portions of 50 cc. each of ether, adding a few drops of
alcohol to facilitate the separation of the two liquids. Combine
the ether extracts in an:other separatory funnel, and wash the
ether solution first with 20 cc. of an aqueous solution of sodium
hydroxide (4 in 1,000), then with 20 ce. of an aqueous solution
of sodium hydroxide (8 in 1,000), and finally with 15 cc. portions
of distilled water until the last washing is not reddened by the
addition of 2 drops of phenolphthalein T .S. Transfer the
ethereal solution to a tared beaker, and rinse the separator with
10 cc. of ether, adding the rinsings to the beaker. Evaporate
355
the ether just to dryness on a water bath, and dry the residue for
30 min. at 100C. Cool the beaker in a desiccator for 30 min.,
and weigh the residue of unsaponifiable matter.
The Pharmacopoeia requires that both cod liver oil and linseed
oil should not contain more than 1.5 per cent and that corn oil
should contain not more than 2 per cent of unsaponifiable matter.
Iodine Value.-The iodine value, or number, is the number of
grams of iodine absorbed by 100 Gm. of oil, jat, wax, or other substance under specified conditions. This value is a quantitative
measure of the proportion of unsaturated fatty acids present,
both free and combined as esters, which have the property of
absorbing iodine.
The determination of the iodine number of fats and oils is
important, since it serves to characterize them and to indicate
whether they are pure or admixtures. The so-called drying oils,
such as linseed oil, and the fish oils, such as cod liver oil, have
very high iodine numbers, usually above 120, since they contain
a large proportion of unsaturated fatty acids; the non-drying oils,
such as olive oil and almond oil, have relatively low iodine
numbers, below 100; and the semidrying oils, such as cottonseed
oil and sesame oil, have intermediate iodine values, that is,
between 100 and 120. In the case of the animal fats, the iodine
number is not very high, usually being less than 90. The determination of the iodine number, therefore, not only serves as an
aid to the identification of known oils, but it also serves to indicate in a definite manner the class to which an unknown fat or
oil belongs. Furthermore, when the iodine number is considered
in conjunction with the saponification value of a fat or oil, it
serves as a means of detecting adulteration, and frequently it
indicates the nature of the adulterant; e.g., olive oil might be
adulterated with cottonseed oil without changing -the saponification value appreciably, but the iodine number of the olive oil
would be increased. Again, castor oil might be adulterated
with olive oil without changing the iodine number greatly, but
the saponification value of the castor oil would be increased.
Several methods have been developed for the determination
of the iodine number of fats and oils. These methods are
generally designated by the name of their originators, as, for
example, the Hubl, Hanus, and Wijs methods. The method
356
QUANTITATIVE PHARMACEUTICAL CHEMIS7'RY
given in the Pharmacopoeia is essentially that of Hanus. In this

method, the following solutions are required:
1. Iodobromide Test Solution.-This solution is prepared as
follows:
"Dissolve 13.2 Gm. of reagent iodine in 1,000 cc. of glacial acetic acid
with the aid of gentle heat if necessary. Cool the solution to 25C. and
determine the iodine content in 20 cc. by titration with tenth-normal sodium
thiosulfate, then add to the remainder of the solution a quantity of bromine
chemically equivalent to that of the iodine present. Preserve in glassstoppered bottles, protected from light."
No difficulty will be experienced in dissolving the iodine, as a

rule, if the glacial acetic acid contains less than 0.5 per cent of
water. The concentration of this solution is determined by
titration with 0.1 N sodium thiosulfate (see page 164). Thus,
if 20.5 cc. of the iodine solution requires 20 cc. of 0.1 N sodium
thiosulfate, each cubic centimeter of the iodine solution must
contain
~~5
X 0.012692 = 0.01238 Gm. iodine, and the remain-
ing 980 cc. of iodine test solution would contain 980 X 0.01238 =
12.13 Gm. of iodine. Since 126.92 Gm. of iodine-is equivalent to
79.92 Gm. of bromine, the quantity of bromine equivalent to
the iodine contained in the solution may be calculated.
X :12.13 ::79.92 :126.92
X = 7.64 Gm. bromine
Since bromine has a specific gravity of about 3.1, 7.64 Gm. of

bromine is equivalent to 7.64/3.1 = 2.5 cc. in round numbers.
Inasmuch as a slight excess of iodine does no harm, the addition
of 2.25 cc. of bromine to the iodine solution would be sufficient
in this case.
The bromine may be drawn into a measuring pipette by means
of a rubber bulb and then added to the cool iodine solution.
(Never draw bromine into a pipette by means of the mouth.)
2. Potassium iodide test solution prepared by dissolving 16.5
Gm. of potassium iodide in sufficient water to make 100 cc.
3. 0.1 N Sodium thiosulfate solution prepared and standardized
as directed on page 158.
4. Starch test solution prepared as follows:
357
Triturate 1 Gm. of arrowroot starch with 10 cc. of cold distilled water,

add sufficient boiling distilled water, with constant stirring, to make 200 cc.,
then boil the mixture until a thin translucent fluid is obtained. The solution
must be freshly prepared and filtered, if necessary.
Exercise 103
Object.-To Determine the Iodine Value of Olive Oil.

Materials Required.-About 1 Gm. of olive oil.
50 cc. of iodobromide test solution.
60 cc. of potassium iodide test solution.
About 100 cc. of 0.1 N sodium thiosulfate solution.
Starch test solution.
Procedure.-1. "Introduce about 0.8 Gm. of a solid fat or about 0.3 Gm. ~
of an oil, accurately weighed, into a glass-stoppered flask or bottle of 250-cc.
capacity, dissolve it in 10 cc. of chloroform, add 25 cc. of iodobromide T.S.,
accurately measured from a burette or pipette, stopper the vessel securely,
and allow it to stand for thirty minutes t in a cool place protected from light."
Only 0.3 Gm. of olive oil is used in this determination, as its

iodine value is comparatively high (79 to 90).
The Pharmacopoeia directs that 0.8 Gm. be taken for solid
fats, as their iodine values are low as compared with those of the
fatty oils, the iodine value of oil of theobroma, for instance, being
33 to 38.
The oil is soluble in chloroform, and since' iodine and iodine
bromide are also soluble in this solvent, it serves as a good
reaction medium.
The oil upon standing in contact with the iodobromide test
solution absorbs iodine. Iodine, itself, is absorbed very slowly
by the oil, but iodine is absorbed readily from bromine containing
solutions, probably through a reaction of iodine bromide, IBr,
wherein the bromine functions as a catalytic agtlnt. The
unsaturated acids of the oleic and linoleic series present in the
olive oil, as well as their glyceryl esters, absorb iodine to form
addition products. Thus oleic ~cid C 17H ssCOOH takes up 2
atoms of iodine and. forms the addition product di-iodo-stearic
acid C 17HssI 2COOH. The mixture, which is directed to be placed
... 0.15 to 0.18 Gm. of linseed oil, 0.18 to 0.2 Gm. of cod liver oil, 0.8 to 1.0
Gm. of oil of theobroma.
t Allow to stand for one hour in the case of castor oil, cod liver oil or
linseed oil.
358
in a glass-stoppered flask (Fig. 61) to prevent the escape of iodine

and bromine vapors, should also be protected from light to
prevent other side reactions from taking place in which iodine is
consumed. If after standing for Y2 hr. the solution is no longer
colored brown, the amount of iodine present was insufficient to
saturate the oil.
2. "Then add in the order named 30 cc. of potassium iodide T.S. and
100 cc. of distilled water, and titrate the liberated iodine with tenth-normal
sodium thiosulfate, shaking thoroughly after each addition of thiosulfate.
When the iodine color becomes quite pale, add 1 cc. of starch
T.S. and continue the titration with thiosulfate until the blue
color is discharged."
Upon the addition of the potassium iodide solution, the iodine bromide reacts with the K1,
liberating iodine until all of the bromine is used up:
K1
+ 1Br~KBr + 12
An excess of K1 is added to insure the complete

removal of the free bromine and to prevent the
precipitation of the iodine in the ~queous solution.
The sodium thiosulfate reacts with the excess iodine ~s follows:
FIG. 61.Iodine titration flask.
2Na 2S20a
+ 12~2NaI + Na2S406
3. "Carry out a blank test at the same time .with the same quantities of
chloroform and iodobromide solution, allowing it to stand for the same length
of time and titrating as directed. The difference in the number of cubic
centimeters of thiosulfate consumed by the blank test and the actual test,
multiplied by 1,269 and divided by the weight of sample taken gives the
Iodine Value.
"NOTE.-If the number of cubic centimeters of tenth-normal sodium
thiosulfate consumed in the actual test is less than 60 per cent of the quantity
consumed in the blank test, the determination must be repeated, using a
smaller amount of the material being assayed."
The blank test, when carried out under the same conditions
as the actual test, corrects for the presence of impurities in the
reagents, changes in volume at different temperatures, etc.,
and makes it unnecessary to know the exact normality Of the
iodobromide test solution. The blank test should be carried out
359
in a manner exactly duplicating that used in the actual test in so

far as possible, especially in the measurement of the iodobromide
test solution, which has a high coefficient of expansion. It is
required that the determination be repeated if the number of
cubic centimeters of 0.1 N Na 2S20a consumed in the actual test
is less than 60 per cent of the quantity consumed in the blank
test to insure the presence of an excess of iodine.
If in the blank test 25 cc. of 0.1 N Na2S20a are consumed, and
in the actual test 5 cc. are required, the calculations may be
made as follows: Since 1 cc. of 0.1 N Na 2S20a is equivalent to
10
~6i9000
, =
0.01269 Gm. iodine, then 25 - 5 = 20 cc. 0.1 N
Na 2S20a equivalent to the iodine absorbed by the oil, and the

weight of iodine which would be absorbed by 100 Gm. of the oil
.
1 t 20 X 0.01269 X 100
20 X 1.269 - 846 th . d'
IS equa 0
0.3
or
0.3
- . , e 10 me
number of the sample of olive oil.
TABLE LIV.-OFFICIAL SUBSTANCES WITH THEIR IODINE NUMBERS
Substance
Iodine Number,
Official Requirement
U.S.P.
Acid, oleic. . . . . . . . . . .
. ......... .
85 to 95
Acid, stearic ...... " . . .. . ....... . Not more than 4
Castor oiL.... . .. . .. . ............ .
83 to 88
Chaulmoogra oil. .................... .
93 to 104
Cod liver oil. ............... , . .. ..
145 to 180
128 to 180
Cod liver oil, non-destearinated. .. . .. .
Corn oiL.... . . . . . . . . . .. .... . ..... .
112 to 128
105 to 114
Cottonseed oil. . . . . .. . .............. .
Ethyl chaulmoograte ................. .
90 to 100
Expressed oil of almond .............. .
93 to 100
Lard .................... .
46 to 70
Linseed oil. . . . . . . . . . . . . . . . .. . ...... . Not less than 170
Oil of theobroma. . . . . . .. . .......... .
35 to 40
Olive oil. ........................... .
79 to 88
Prepared suet. . . . . . .. . ............. .
33 to 48
Soap, hard (acids from) .......-;-....... .
83 to 92
Soap, soft (acids from) ............... . Not less than 170
Wool fat ........................... .
18 to 36
N.F.
Croton oil. ......................... .
104 to 110
Oil of sesame ........................ .
103 to 115
360
QUANTITATIVE PHARMACEUTICAL CHGMISTRY

1. Define the term iodine value.

2. Indicate by structural formulae how iodine may adci on to an unsaturated fatty acid.
3. How maya determination of the iodine number be used to supplement
the information relative to the nature and purity of an oil gained through
a determination of the saponification value?
4. What functions do the bromine, acetic acid, chlorofo:em, and potassium
iodide serve in the determination of the iodine number of an oil?
5. Given an unknown sample of a fatty oil. What observatioIis, tests,
and determinations would you perform in order to establish the identity
and purity of the oil?
CHAPTER XXIII
Volatile oils, also known as ethereal oils or essential oils and, in
some cases, as essences, are generally complex products composed
of mixtures of compounds of widely variant chemical characteristics. The most important chemical components of the
official volatile oils are:
1. Hydrocarbons, occasionally of the aliphatic series, such as
heptane, myrcene, and various paraffins; but more often of the
aromatic series, e.g., pinene, camphene, limonene, bornylene,
fenchene, dipentene, sylvestrene, and phellandrene.
2. Alcohols, present both in the free state and in combination
with acids as esters. The alcohols most generally found in the
official volatile oils are linalool, geraniol, citronellol, terpineol,
borneol, menthol, and santalol.
3. Aldehydes, such as benzaldehyde, cinnamic aldehyde, salicyl
aldehyde, citral, and citronellal.
4. Ketones, the most important being camphor, carvone,
fenchone, thujone, and menthone.
)
5. Phenols, e.g., anethol, eugenol, carvacrol, safrol, chavicol,
and thymol.
6. Acids, sometimes present in the free state in small quantities, those occurring most commonly being acetic, propionic,
butyric, valerie, benzoic, cinnamic, and hydn>cyanic acids; but
more often occurring in combination with the alcohols mentioned under 2 as esters, e.g., linalyl acetate, bornyl acetate, and
menthyl acetate.
I
7. Sulfur compounds, such as\. the allyl thiocyanate, found in
mustard oil.
The analysis of volatile oils for the purposes of determining
their purity and value is based on the measurement of certain
physical characteristics, on the quantitative estimation of certain
components as alcohols, phenols, esters, and aldehydes, and on
361
362
the qualitative tests for the various substances which are commonly employed as adulterants. Only the official quantitative
procedures will be discussed here.
Methods of General Application.-The purity and quality of
volatile oils may be judged to some extent by their appearance,
odor, color, etc.; but the information gained from the determination of the specific gravity, rotatory power, refractive index,
solidifying point, solubility, and behavior on distillation is of
much greater importance.
Specific Gravity.-The specific gravity of a volatile oil may be
determined with the Westphal balance or pycnometer, the
latter being the more accurate method of the two, and expressed
as the ratio of the weight of the volume of oil to that of an equal
volume of pure water when both are determined at 25C. (see
page 217).
The specific gravities of the official volatile oils vary approximately between 0.84 and 1.2. Those oils which are lighter
than water are usually rich in hydrocarbons, alcohols, esters,
aldehydes, and ketones, such as oil of orange, caraway, coriander,
lemon, turpentine, and rosemary. Oils the spt;;_cific gravities of
which approach or exceed 1.0 usually contain chiefly phenols,
phenolic derivatives, or certain esters, e.g., oil of anise'lcinnamon,
clove, sassafras, and mustard.
The specific gravity of any volatile oil is not absolutely constant, since it is influenced by such factors as the matl,J.rity of the
plant from w!ich the oil is obtained, as well as the method of
preparation, purification, and age of the oil.
Rotatory Power.-The rotatory power of a volatile oil is
generally measured with a Laurent half-shadow polarimeter,
according to the procedure described on page 247, using sodium
light and a tube 10 cJ?1. long, but for highly colored oils, tubes 5
or even 2.5 cm. long may be used. The observation of the
optical activities of the official essential oils should be made at
25C. Slight deviations from this temperature do not greatly
affect the rotatory power of a volatile oil, except in the case of oil
of lemon and oil of orange.
The rotatory power of some of the volatile oils varies within
relatively wide limits. This determination should never be
omitted in their examination, however, since it frequently serves
363
as a valuable means of detecting adulteration with inactive substances, such as alcohol t or with substances of different rotatory
power from that of the oil being examined, e.g., oil of lemon
(+57 to +64) adulterated with oil of turpentine (about +25 to
-40).
Refractive Index.-The refractive index of a volatile oil is
most commonly determined by means of an Abbe refractometer,
as described on page 240. The measurement of the refractive
indices of all of the official volatile oils should be performed at
20C. The index of refraction does not vary greatly with
different official volatile oils, the values being between about
1.46 and 1.61 at 20C. In some cases, this determination may,
however, serve for the detection of extraneous matter.
Congealing Point.-The congealing point of a volatile oil is
determined by the method described on page 231. Most essential oils solidify only at low temperatures, consequently in
practice, this determination is carried out with only a few oils,
such as oil of anise and oil of fennel, which contain large amounts
of the readily crystallizable constituent anethol. The higher
the congealing point of these oils the more they are valued.
An abnormally low congealing point of an essential oil indicates
the partial removal of the characteristic constituent for which
the oil is valued or the addition of extraneous matter, such as
alcohol.
Distilling Point.-The distilling point of a volatile oil is
determined by the general method described on page 234.
Volatile oils which are composed of mixtures of hydrocarbons,
alcohols, esters, etc., do not have a fixed boiling point but boil
between certain limits of temperature, frequently separated
widely. Consequently, the official standards usually designate
the temperature or range of temperature at which a definite
percentage of the oil distils; e.g., 90 per cent of oil of turpentine
should distil between 154 and 170C., and less than 10 per cent of
oil of dwarf pine needles should aistil below 1650.
Fractional Distillation.-Fractional distillation is resorted to
occasionally in the official tests for purity of volatile oils. It
serves either to' separate the various components of volatile oils
or to detect adulteration. Thus, alcohol and petroleum ether,
both of which boil below 1000., may be separated and identified
364
in this way. The addition of oil of turpentine and other adulterants may be detected by collecting various fractions of the oil
and determining the rotary power and refractive index of
each fraction; e.g., see Oil of Orange, U.S.P., page 250.
Solubility.-The volatile oils are generally soluble in organic
solvents, such as absolute alcohol, ether, chloroform, benzene,
carbon disulfide, etc. They dissolve more or less readily in
dilute alcohol according to the nature of their components. Oils
containing a large percentage of oxygenated substances often
produce turbid solutions with petroleum ether or carbon disulfide due to the separation of water, small quantities of which
are dissolved in such oils.
Practically all of the volatile oils exhibit almost constant
solubility in 90, 80, or 70 per cent alcohol. Consequently, the
test of the solubility of a volatile oil in dilute alcohol frequently
gives valuable data relative to its purity, since the commonly
used adulterants, oil of turpentine, petroleum oils, and fatty
oils, are but slightly soluble in 80 or 70 per cent alcohol.
Assay for Ester Content.-A number of the official volatile
oils are evaluated on the basis of their ester GOntent. The esters
found are mostly the acetates of alcohols of the formula C lOH 170H,
e.g., borneol, geraniol, linalool, and terpineol; C 1QH 190H, such
as menthol or citronellol; or C 15H 2SOH, e.g., santalol. When
these esters are refiuxed with alcoholic potassium hydroxide,
they are saponified and yield the free alcohol and the potassium
salt corresponding to the acid component of the ester.
The number of milligrams of KOH required to saponify 1 Gm.
of the oil represents the ester value or ester number of the oil.
The ester number for any given oil may be subject to considerable
variation, but the minimum requirement set by the official
standards must be complied with when the oil is represented as a
product conforming to the U.S.P. or N.F. standards. The
determination of the total esters when taken in conjunction with
the official tests for purity serves to detect adulteration
and to establish the quality of those oils valued for their ester
content.
The ester values of oils which contain appreciable amounts of
aldehydes or phenols cannot be estimated accurately by saponification with an alkali, since variable amounts of the latter are
365
consumed by reaction' with the aldehydes and phenols, or their

decomposition products.
Exercise 104
Object.-To Prepare 0.5 N Alcoholic Potassium Hydroxide.

Materials Required.-2.5 Gm. of lead acetate.
1,500 cc. of alcohol.
40 Gm. of potassium hydroxide.
Procedure.-l. "Dissolve 2.5 Gm. of lead acetate in 5 cc. of distilled
water, add the solution to 1000 cc. of alcohol contained in a glass-stoppered
bottle and mix thoroughly. Dissolve 5 Gm. of potassium hydroxide in
25 cc. of warm alcohol, cool the solution, and add it slowly, without stirring,
to the alcoholic solution of lead acetate. Mter one hour shake the mixture
vigorously, allow it to stand over night, decant the clear liquid and recover
the alcohol by distillation."
The potassium hydroxide converts any acids present into nonvolatile salts and causes the polymerization of aldehydes. The
lead acetate is decomposed by the KOH with the formation of
lead oxide and potassium acetate. The lead forms insoluble and
non-volatile compounds with any sulfur compounds present.
The lead oxide carries down the polymerized aldehydes when it
is precipitated so that it is easy to decant the clear supernatant
liquid.
2. "Dissolve about 35 Gm. ot potassium hydroxide in 20 cc. of distilled
water and add sufficient aldehyde-free alcohol to make 1000 cc. Allow the
solution to stand in a tightly-stoppered bottle (using either glass or rubber
stopper) for twenty-four hours. Then quickly decant the clear supernatant liquid into a bottle provided with a well-fitting rubber-stopper and
standardize as follows:"
Traces of aldehydes remaining in the alcoholic potassium

hydroxide solution are polymerized and settle out when the
mixture is allowed to stand. The rubber stopper used should
be washed with a small quantity of the alkaline alcoholic solution
before it is inserted into the -mouth of the bottle to dissolve
sulfur and any other material which might affect the concentration of the solution.
3. "Measure accurately, from a burette, about 25 cc. of half-normal
hydrochloric acid. Dilute with 50 cc. of distilled water, add 2 drops of
phenolphthalein T.S., and titrate with the alcoholic potassium hydroxide
366
solution until a permanent, pale pink color is produced.

normality."
Calculate the
The titration is carried out as described on page 81. The

prepared solution should be preserved in tightly stoppered bottles
to prevent changes in the concentration of alkali through absorption of CO 2 Well-fitting rubber stoppers may be used. It
should also be protected from light to prevent the development
of reddish colored compounds which interfere with the correct
determination of the end point during titration. The -solution
should be restandardized whenever it is used, and it should
always be measured at the same temperature at which it was
standardized, since alcohol has a high coefficient of expansion.
Exercise 106
Object.-Assay of Oil of Peppermint for Total Esters.

Materials Required.-10 cc. of oil of peppermint.
25 cc. of 0.5 N alcoholic KOH.
About 25 ce. of 0.5 N H 2SO.
Procedure.-l. "Place 10 cc. of oil of peppermint in a tared, 125 cc.
Erlenmeyer flask, and weigh accurately. Add 25 cc. oJ-half-normal alcoholic
potassium hydroxide, connect the flask with a reflux condenser, and boil
the mixture on a water bath for one hour."
Instead of weighing the oil directly in a previously tared

Erlenmeyer flask, it may be weighed in a weighing bottle, and the
weighing bottle with its contents may then be transferred into the
Erlenmeyer flask. The esters of oil of peppermint, which consist chiefly of menthyl acetate, are saponified by the alkali when
the mixture is refluxed, forming free menthol and potassium
acetate. Alcoholic potassium hydroxide is used because the
oil is soluble in strong alcoholic solution.
A -(}--G--CH, + KOH____'~H + CH,COOK

o
'\-
'
2. "Allow the mixture to cool, disconnect the flask from the condenser,
and titrate the excess of alkali with half-normal sulfuric acid, using 10
367
drops of phenolphthalein T.S. as the indicator. Subtract the number of

cc. of half-normal sulfuric acid required for neutralization from the 25 cc.
of half-normal alcoholic potassium hydroxide taken, multiply the difference
by 9.912, and divide this product by the weight of oil of peppermint taken,
the result shows the per cent of esters calculated as menthyl acetate."
The 0.5 N acid neutralizes the excess 0.5 N alkali:
The difference between the number of cubic centimeters of 0.5 N

KOH added and the number of cubic centimeters of 0.5 N H 2S0 4
required in the titration represents the alkali consumed in the
saponification, of the esters present in the oil; e.g., if a 9 Gm.
sample of the oil refluxed with 25 cc. of 0.5 N KOH required 20.5
cc. of 0.5 N H 2S0 4 to neutralize the excess alkali, the amount of
standard alkali solution consumed in the saponification of the
esters present in the oil would be 25 - 20.5 = 4.5 cc. The percentage of total esters present in the oil calculated as menthyl
acetate is found as follows: 4.5 X 9.912 = ~.96 per cent menthyl
9
acetate. The factor 9.912 is derived from the molecular weight of
menthyl acetate, 198.24. Thus, 1,000 cc. of N KOH is equivalent
to 198.24 Gm. of menthyl acetate, and 1 cc. of 0.5 N KOH is
equivalent to 2
~8i~0400
0'.09912 Gm. menthyl acetate.
This
equivalent is multiplied by 100 so that the factor 9.912 when

multiplied by the number of cubic centimeters of 0.5 N alkali consumed in the assay and divided by the weight of the sample of
oil used will give the percentage of esters directly.
1. What effect would the presence of free acids in th!l oil have upon the
result of the determination of total esters?
2. Why is an alcoholic rather than an aqueous solution of KOR used to
saponify the esters?
\0
3. What factor would be necessary for the ester in order to obtain the
per cent of total esters directly as described above if the chief ester present
in an oil was bornyl acetate; linalyl acetate? Show how these factors are
derived.
.
4. Note the equivalents given in the following table for linalyl acetate in
oil of lavender, U.S.P., and in oil of bergamot, N.F. Which is correct?
368
TABLE LV.-OFFlCIAL VOLATILE OILS ASSAYED FOR THEIR ESTER CONTENT

Oil
Amount Equivalent of
used,
1 cc. of 0.5 N
Gm.
KOH,Gm.
per cent
U.S.P.
Methyl salicylate ......
0.07603
Methyl salicylate
C.H4(OH)CO.CH. = 98
Oil of dwarf pine

needles .............
10
0.09811
Oil of lavender ........
0.09811
Oil of peppermint .....
10
0.09912
Oil of rosemary ........
10
0.09811
Bornyl acetate
CloHI7C.H.02 =
Linalyl acetate
C IOH I7C.H.O. =
Menthyl acetate
CloHuC.HaO. =
Bornyl acetate
C IOH I7C.HaO. =
N.F.
Oil of bergamot .......
0.09808
5
30
5
2.5
Linalyl acetate
C1oH'7C.H.O. = 36
Assay for Alcohol Content.-The alcohols present in volatile

oils, such ~s menthol and borneol, occur both free and
combined as esters. Usually, the, <letermination of
the total alcohol, occurring free and combined, is
sufficient to establish the purity and valu~. of an oil
with respect to its content of alcoholic constituents.
The total alcohols present in any given oil are
determined by transforming the free alcohols into the
corresponding acetates by boiling the oil with acetic
anhydride in an acetylization flask (Fig. 62) and then
determining the saponification value of the acetylized
product. When the alcohol occurs partly free and
partly combined in the form of an ester, a correction
factor mus~ be employed. The method of determining the total alcohol content of volatile oils is
illustrated in the following exercise:
FIG. 62.Acetylization flask
wi th air
condenser.
Exercise 106
Object.-Assay of Oil of Peppermint for Total

Menthol.
Materials Required.-lO cc. of oil of peppermint.

10 cc. of acetic anhydride.
369
1 Gm. of anhydrous sodium acetate.

3 Gm. of monohydrated sodium carbonate.
About 0.2 Gm. of fused calcium chloride.
50 cc. of 0.5 N alcoholic ROll.
50 cc. of 0.5 N H 2S0 4
An acetylization flask of about 100 cc. capacity with an air condenser
about 100 cm. long.
Procedure.-l. "Place 10 cc. of Oil of Peppermint in an acetylization
flask of 100 cc. capacity, add 10 cc. of acetic anhydride and 1 Gm. of
powdered anhydrous sodium acetate. Boil the mixture gently for one hour,
cool, disconnect the flask from the condenser, transfer the mixture to a
small separator, rinsing the acetylization flask with three successive 5 cc.
portions of warm distilled water, and add the rinsings to the separator."
The acetylization flask should be supported on an asbestos

board while heating to prevent decomposition of the oil. The
acetic anhydride reacts with the menthol, forming menthyl
acetate and acetic acid:
C-CH3
OH+O
'\,
-O-C-CH 3 + CHaCOOH
G-CH s
,f'
Anhydrous sodium acetate is added to absorb any traces of

water liberated in the side reaction:
.
6
-OH
0
~
+ H0-C--CHs-
'\,
O-G-CHs + HOH
2. "When the liquids have completely separated, reject the aqueous layer,
and wash the remaining oil with successive portions of sodium carbonate
T.S., diluted with an equal volume of distilled water, until the washing is
alkaline to 2 drops of phenolphthalein T.S. Dry the resulting Oil with
anhydrous sodium sulfate (prepared by drying sodium sulfate to constant
weight at 110C. and powdering), and filter it."
370
The oil is washed with sodium carbonate test solution (12.5 Gm.
of Na2,C0 3 .H2,O ,in sufficient distilled water to make 100 ce.) to
neutralize the excess acetic acid. The washed oil is then dried
over anhydrous sodium sulfate to remove the small amounts of
alkaline sodium carbonate solution which it contains.
3. "Transfer 5 cc. of the dry acetylilled oil to a tared, 100-cc. Erlenmeyer
flask, note its exact weight, add 50 ce. of half-normal alcoholic potassium
hydroxide, connect the flask with a reflux condenser, and boil the mixture
on a water bath for one hour."
The 0.5 N KOH saponifies the menthyl acetate, forming menthol and potassium acetate in the s~me manner as in the assay for
total esters. The liberated menthol dissolves in the alcoholic
solution.
4. "Allow the mixture to" cool, disconnect the flask from the condenser,
and titrate the excess of alkali with half-normal sulfuric acid, using 10 drops
of phenolphthalein T.S. as the indicator. Calculate the per cent of menthol
by the following formula:
Per cent of total menthol in the Oil tested
!(~ ~8~~21)
_x [1 - (E X 0.0021)].
A is the result obtained by subtracting the number of cc of half-normal

sulfuric acid required in the above titration from the numb~ of cc. of halfnormal alcoholic potassium hydroxide originally taken, B is the weight of
acetylized oil taken, and E is the percentage of esters calculated as menthyl
acetate (CloH19.C2Ha02)."
The number of cubic centimeters of 0.5 N KOH consumed in

the saponification of a definite weight of the acetylized oil is
equal to the difference between the number of cubic centimeters
of 0.5 N KOH added and the number of cubic centim'eters of 0.5 N
H 2S0 4 required in the residual titration. The formula used in
the calculation is derived as follows: The molecular weight of
menthol is 156.16. One cubic centimeter of 0.5 N KOH is
equivalent to 2
~6i~0600 = 0.07808 Gm. menthol.
,In order that
the result may be read directly in terms of per cent, the_factor

0.07808 is multiplied by 100, giving 7.808. B - (A X 0.021)
corrects for the weight of acetyl radical in the sample of acetylized oil, the factor 0.021 being the amount of acetyl radical
equivalent to 1 cc. of 0.5 N KOB. The latter is obtained from
371
198.17 (moleculal' weight of menthyl acetate) - 156.16 (molecular weight of menthol) = 42.01 (molecular weight of the acetyl
radical). Therefore, 1,000 cc. of N KOH is equivalent to 42.03
Gm. of acetyl radical, and 1 cc. of 0.5 N KOB is equivalent to
2 X 4;,000
= 0.021 Gm. acetyl radical.
(A X 0.021), therefore,
represents the weight of acetyl radical to be deducted from B, the

weight of acetylized oil taken.
Since the original oil contained some menthol combined as ester
and calculated as menthyl acetate, it would be incorrect to subtract all of the weight of the acetyl radical from B. A correction
for the weight of acetyl radical in the original oil is consequently
applied by multiplying the per cent found from the formula
B
! (~ ~8g.~21) by the correction formula [1 -
(E X 0.0021)].
The factor 0.0021 is the ratio of the weight of acetyl radical to

that of menthyl acetate 42/198.17 = 0.21. If E, the per cent of
total esters determined by the preceding assay, is found to be 5,
then the formula would be [1 - (0.05 X 0.21)] or [1 - (5 X
0.0021)]. Applying this correction, the formula becomes
A X 7.808
B _ (A X 0.021) X [1 - (5 X 0.0021)] = per cent total menthol.
The importance of the correction [1 - (0.05 X 0.21)] may be
seen from the following example: A sample of oil of peppermint
found to contain 5 per cent of total esters calculated as menthyl
acetate and 50 per cent of total menthol as calculated from the
A X 7.808
formula, per cent total menthol = B _ (A X 0.021)' gave the
following percentage of total menthol when the correction was
applied: 50 X [1 - (5 X 0.0021)] = 50 X (1 - 0.0105) = 50 X
0.9895 = 49.475 per cent menthol free and as ~sters. The
correction involves about one-half of 1 per cent menthol or 50 49.475 = 0.525 per cent.
A similar correction is applied in the assay of oil of rosemary,
but none is given in the assay ~f oil of santal, since the santalol
does not occur combined in the form of esters.
Questions and
Problem~
1. Having determined the ester value and 'the percentage of total borneol
present in oil of rosemary, how could the percentage of free borneol be
found? Give an example.
;372
2. Write the reactions which take place in the assay of oil of rosemary for
total borneol.
3. If oil of peppermint containing 7.2 per cent of total esters is found to
contain 51.5 per cent of total menthol (uncorrected), what change in the
result would the correction 1 - (E X 0.0021) cause? Is this correction
important?
TABLE LVI.-OFFICIAL VOLATILE
OILS ASSAYED
FOR THEIR
ALCOHOL
CONTENT
Oil
Amount of
acetylized
oil used,
Gm.
Equivalent
of 1 cc. of
0.5 N KOH,
Gm,
Correction factor
U.S.P.
Oil of peppermint ...
0.07808
1 - (E X 0.0021)
Oil of rosemary .....
0.0771
1 - (E X 0.0021)
Oil of Ban tal. .......
0.1101
None
Menthol
ClOH"OH =50
Borneol
ClOH170H = 10
Santalol
C"H"OH = 90
Assay for Aldehyde Content.-Aldehydes, like ketones, form

addition products with certain reagents, anq,. ..this property is
utilized in their assay. When no other constituents are present which react with the reagent, the assay for aldehydes may be
performed by the sulfite method described under the assay for
ketones (page 375). Thus the cinnamic aldehyde content of the
official oil of cinnamon is determined by this method; the basic
reaction being
/'
CR CH-C
'"
NaO
0
'" /'
+2
SH
/~
Na
0
+ 2NaOH
373
The sulfite addition product dissolves in water leaving the nonaldehyde constituents as a water-insoluble layer. The volume
of this water-insoluble layer is then measured in a cassia flask,
(Fig. 63).
The cassia flask is a glass flask of about 100 cc. capacity with a
long, narrow neck graduated to 10 cc. in 0.1 cc. divisions. In the
assay for aldehydes and ketones by the sulfite method or bisulfite method, the portion of the oil which does not react to form
a water-soluble addition product rises to the surface, and when
the flask is filled, this oily layer rises into the graduated neck
of the cassia flask where its volume can be measured. The volume of the residual oily layer
subtracted from the volume of the sample of oil used
represents the volume of the aldehyde constituents
r
which formed a water-soluble addition product with
the sulfite.
I:
Those volatile oils which contain aldehydes in
very small amounts, such as oil of lemon, or which
contain other constituents that form water-soluble
addition products with sodium sulfite and bisulfite
cannot be assayed accurately by this method. In FIG. 63.Cassia
these cases, the aldehydes are generally determined flask.
by the phenylhydrazine method (see assay of
benzaldehyde, N.F.) or by the hydroxylamine method described
in the following exercise.
Exercise 107
Object.-Assay of Oil of Bitter Almond for Benzaldehyde.

Materials Required.-1 cc. of oil of bitter almond.
3.5 Gm. of hydroxylamine hydrochloride.
100 cc. of 60 per cent alcohol.
Bromphenol blue T.S.
0.5 N alcoholic KOH.
Procedure.-l. "Add 0.1 cc. of bromphenol blue T.S. to 25 cc. of a solution made by dissolving 3.5 Gm. of hydroxylamine hydrochloride in sufficient
60 per cent alcohol to make 100 cc., and titrate with half-normal alcoholic
potassium hydroxide to the production of a greenish-blue color."
Hydroxylamine hydrochloride reacts as a neutral salt when

bromphenol blue is used as the indicator. Consequently, upon
374
titration of the solution of hydroxylamine hydrochloride with

0.5 N KOH, the free Hel present in the solution is neutralized.
If an indicator which changes color on the alkaline side Of the
pH scale, such as phenolphthalein, is used, the end point will be
obtained when the free and combined acid has been neutralized
by the alkali.
2. "Pour this mixture into a flask, fitted with a glass stopper, and containing about 1 Gm. of Oil of Bitter Almond, accurately weighed. Shake
well and then titrate with half-normal alcoholic potassium hydroxide until
the yellow color changes to greenish-blue. Continue shaking and titrating
until the greenish-blue color is permanent. Each cubic centimeter of halfnormal alcoholic potassium hydroxide is equivalent to 0.05303 Gm. of
CsH. CHO."
Aldoximes are formed when a cold aqueous or hydroalcoholic

solution of hydroxylamine hydrochloride is added to an aldehyde
in the presence of an equivalent amount of KOH. The equations
for the reactioJ?s that take place may be represented as follows:
The basic hydroxylamine reacts with the benzaldehyde to form an

unstable addition product which loses water and readily forms the
oxime.
It is apparent from the equations given that the assay is based

upon the determination of the amount of Hel obtained from the
quantity of hydroxylaniine hydrochloride equivalent to the
benzaldehyde contained in the sample.
The HeN present in the oil of bitter almond does not influence
the accuracy of the assay. Hydrocyanic acid is a very weak acid.
In the presence of a strong acid, such as Hel, the common
hydrogen ion represses the dissociation of the weak acid, HeN,
so that at the end point, which in this case is at a pH of about
4.2, none of the HeN enters into the reaction.
375
TABLE LVII.-OFFICIAL VOLATILE OILS ASSAYED FOR ALDEHYDE

CONTENT
Oil
Amount
used
per cent
Method
U.S.P.
Oil of bitter almond ......... 1 Gm. Hydroxylamine
Oil of cinnamon .. 10 cc.
Sulfite
Benzaldehyde = 95
Cinnamic aldehyde = 80
N.F.
Benzaldehyde ....
1 Gm. Phenylhydrazine Benzaldehyde = 85

1. Show how the equivalent factor for benzaldehyde is derived.

2. Why is the titration carried out in a hydroalcoholic solution?
3. Look up the method of assay for benzaldehyde in the National
Formulary and write equations for 'the reactions that take place.
4. Does the presence of HCN' cause an appreciable error in the assay of
oil of bitter almond for benzaldehyde content?
Assay for Ketone Content.-Only two of the official volatile

oils are assayed for their ketone content. In each case, the
determination is made by the so-called sulfite method. This
method is based on the property of ketones to combine with
sodium sulfite to form water-soluble addition compounds.
Exercise 108
Object.-Assay of Oil of Caraway.

Materials Required.-A 100 cc. cassia flask.
10 cc. of oil of caraway.
100 cc. of saturated sodium sulfite solution (40 Gm. in 100 cc.).
100 cc. of sodium bisulfite solution (5 per cent).
Procedure.-l. "Intrqduce exactly 10 cc. of Oil of Caraway, accurately
measured, into a cassia flask, and add 50 cc. of a saturated solution of sodium
sulfite which has been carefully rendered neutral to 2 drops of phenolphthalein T.S. by means of a saturated sodium bisulfite solution. Heat the
flask in boiling water and shake it repeatedly, neutralizing the mixture from
time to time by the addition of a few drops of the saturated sodium bisulfite
solution."
Carvone, the principal ketone present in the oil, reacts with the
sodium sulfite, forming a water-soluble addition product as
follows:
376
This addition product is hydrolyzed in part by the water with the

formation of sodium hydroxide, viz.:
ONa
()<sE~
) ~
ONa
The presence ,of free alkali in the aqueous layer renders other
components of the oil, such as phenols, water-soluble and also
tends to reverse the addition reaction. To pt:event this the
sodium hydroxide is neutralized by the addition of sodium acid
sulfite:
2. "When no coloration appears upon adding a few more drops of phenolphthalein T.S. and heating for 15 minutes, cool the mixture to room temperature, and, when the liquids have separated completely, add sufficient
sodium sulfite solution to raise the lower limit of the oily layer within the
graduated portion of the neck of the flask. Note the volume of the residual
oily liquid. This volume does not exceed 5 cc., indicating the presence in
the Oil of not less than 50 per cent, by volume, of carvone."
The difference between the volume of oil used as the sample and
the volume of the oily layer which remains insoluble represents
the carvone which dissolved in the aqueous layer. If drops of
oil adhere to the walls of the fiask,-they may be made to rise into
the neck by gently tapping and rotating it. If the residual
liquid measures 4.5 cc., the percentage of carvone by volume in
the oil would be 10
~O 4.5
X 100 = 55 per cent.
The only other official oil evaluated for its ketone content is oil
of spearmint. It is assayed in exactly'the same way as is oil of
377
caraway, and it is required to contain not less than 50 per cent by

volume of carvone.
'
1. Ketones may also be estimated in volatile oils by the use of sodium
bisulfite alone. Write the reaction between carvone and sodium bisulfite.
2. What substances, if present, in the oil would cause an error in the
results of the sulfite method of assay for. ketones?
3. How could the carvone be isolated from the aqueous solution?
Assay for Phenol Content.-Volatile oils which contain phenols

when shaken with solutions of sodium or potassium hydroxide
diminish in volume due to the ready solubility of the phenol
constituents in alkali; the non-phenolic portion of the,oil remains
undissolved.
Use is made of this property of phenols in the official assay
methods. In these determinations a flask with a graduated neck
like that employt'!d in the estimation of aldehydes (see page 373)
is used. In this flask, an accurately measured volume of the oil is
shaken vigorously with a portion of the caustic alkali, and a
further quantity of alkali is then added to bring the undissolved
oil into the graduated neck of the flask. When the liquids have
separated and become clear, the volume of the oil insoluble in
alkali is read off, the volume of the phenol being determined by
difference.
Exercise 109
Object.-Assay of Oil of Clove.

Materials Required.-l0 cc. of oil of clove.
100 cc. of KOH solution (6.5 Gm. in 100 cc.).
A cassia flask.
Procedure.-l. "Place 10 cc. of Oil of Clove, measured from a pipette,
in a 100-cc. cassia flask, add 50 cc. of normal potassium hydroxide solution,
shake the mixture for five minutes, and heat it for ten minutes in a bath
containing boiling water. Remove it from the bath, and cool to room
temperature. "
The KOH reacts with the phenol, eugenol, as follows:

-OH
-OCH3
-CH 2-CH
+KOH~
CH 2
-OK
O-OCH3
-CH2-CH
+HOH
CH2
378
The mixture is heated on a water bath for 10 min. to saponify

any aceteugenol present.
2. "When the liquids have separated completely, add sufficient n~rmal
potassium hydroxide to ra.ise the lower limit of the oily layer within the
graduated portion of the neck: the volume of the oily layer does not exceed
1.8 cc., indicating the presence in the Oil of not less than 82 per cent by
volume-of eugenol (C lOH ,202)."
The potassium eugenol compound dissolves in the water leaving

undissolved hydrocarbons, ketones, etc., which rise into the
graduated neck of the flask and are measured. If stronger solutions are employed, thc results obtained are too high, since the
strong alkali exercises some solvent action on the non-phenolic
constituents, partidularly on oxygenated compounds. The volume of the undissolved liquid subtracted from the volume of oil
used as the sample gives the volume of eugenol which dissolved in
the alkali; e.g., if a 10 cc. sample of oil yields 1.6 cc. of residual
liquid, then 10 - 1.6 = 8.4 cc. eugenol dissolved, and the percentage of eugenol in the sample is
~t
X 100 = 84 per cent.

1. Is the assay of phenols, as given above, an exact' meth~d?
2. How could the eugenol be liberated from the alkaline solutidn?
3. If a part of the eugenol is present combined as esters, is that part
estimated in the determination?
I
TABLE LVnI.-OFFICIAL VOLATILE OILS ASSAYED FOR PHENOL CONTENT
Amount
used, cc.
Alkali
used
per cent
clove, , , , , .. '.' ......
10
KOH
Eugenol C lOH ,2 0 2 = 82
myrcia ..............
pimenta. , ..........
thyme ... , ..........
10
10
10
KOH
KOH
KOH
Phenols = 50 to 60
Eugenol,C ,o H 120 2 = 65
Phenols = 20
Oil
U.S.P.
Oil of
N.F.
Oil of
Oil of
Oil of
Assay for Hydrocyanic Acid Content.-Only one official volatile oil is assayed for its hydrocyanic acid content, namely, oil of
bitter almond.
379
Exercise 110
Object.-Assayof Oil of Bitter Almond for Hydrocyanic Acid.

Materials Required.--o.75 Gm. of magnesium sulfate.
Potassium chromate indicator (10 Gm. in 100 cc. of water).
0.1 N silver nitrat~.
Procedure.-l. "Dissolve 0.75 Gm. of magnesium sulfate in 45 cc. of
distilled water, add 5 cc. of half-normal sodium hydroxide and 2 drops of
potassium chromate T.S., and titrate the solution with tenth-normal silver
nitrate to the production of a permanent reddish tint."
The magnesium sulfate reacts with the sodium hydroxide,

forming magnesium hydroxide and sodium sulfate:
MgS0 4
+ 2NaOH~Mg(OH)2 + Na 2S04
The resulting mixture is titrated with 0.1 N silver nitrate to the

appearance of a permanent end point to correct for the presence
of impurities in the reagents used which would react with the
silver nitrate in the next step of the assay.
2. "Pour this mixture into a 100 cc. flask containing about 1 Gm. of
Oil of Bitter Almond, accurately weighed, mix well, and titrate again with
tenth-normal silver nitrate until a red tint, which does not disappear on
shaking, is reproduced. Conduct this titration as rapidly as possible.
Each cubic centimeter of tenth-normal silver nitrate corresponds to 0.002702
Gm. of HCN."
The freshly precipitated Mg(OH)2 splits up the cyanhydrin

present in the oil and at the same time furnishes a white background against which the end point of titration may be observed.
The reacti~n may be represented as follows:
H
20
-C-OH
"C=N
+ Mg(OH),-.
H
/
C~
0
+ Mg(CN)2 + 2HOH
380
Upon titration with 0.1 N silver nitrate, silver cyanide is form~d:

Mg(CN)2
+ 2AgNOa~2AgCN + Mg(NO a)2
When all of the alkali cyanide present in the mixture is consumed

in the reaction with the silver nitrate, the end point is shown by
the formation of the red-colored precipitate of silver chromate:
K2Cr04
+ 2AgNOa~Ag2Cr04 + 2KNOa
The flask should be shaken continuously during the titration to

bring the oil into intimate contact with the mixture. The titration is directed to be conducted as rapidly as possible, because
HCN decomposes in the presence of water and alkalies with the
formation of ammonia, formic acid, etc.
Each cubic centimeter of 0.1 N AgNO a consumed in the final
titration is equivalent to 10 ~~~ooo = 0.002702 Gm. HCN where
27.02 is the molecular weight of hydrocyanic acid.
1. Why is the magnesium sulfate added?

2. From the quantities of MgSO. and NaOH used, cal~ulate whether
any NaOH as such will be present in the reaction mixture.
3. Indicate by reactions how HeN may be hydrolyzed in the presence
of alkalies with the formation of formates and ammonia.
Assay of Oil of Chenopodium for Ascaridol Content.-The

principal constituent of oil of chenopodium from a pharmaceutical
standpoint is ascaridol, an organic peroxide, which is valued for its
anthelminthic properties. Various chemical and biological
methods of evaluating the oil have been proposed, but the official
method, which is based on the solubility of ascaridol in acetic
acid, is probably as good as any, although it should be regarded as
approximate only.
Exercise 111
Object.-Assay of Oil of Chenopodium.

Materials Required.-lO cc. of oil of chenopodium.
60 cc. of glacial acetic acid.
A cassia flask.
381
Procedure.-l. "Place 10 cc. of Oil of Chenopodium, measured from a

pipette, in a 100 cc. cassia flask, add 50 cc. of a solution of acetic acid, made
by diluting 60 cc. of glacial acetic acid with distilled water, to measure
100 cc."
The ascaridol in the oil dissolves in the 60 per cent acetic acid.
The exact nature of the reaction that occurs is not known, but it
may be represented as follows:
{D
I
~
I-O-C-CHs
+ 2CH'COOH~O + 2HOH + 0,
Any variation in the strength of the acetic acid employed causes

variable amounts of the constituents of the oil (decomposition and
reduction products of ascaridol) to dissolve in the acetic acid;
consequently, the acetic acid solution used must be of exactly
60 per cent strength.
2. "Shake the mixture continuously and vigorously for a period of five
minutes and add sufficient of the acetic acid solution to raise the lower limit
of the oily layer within the grad~ated portion of the neck. When the
liquids have completely separated so that a sharp meniscus is discernible,
note the volume of the oily layer, at 250. It measures not more than 4 cc.
and not less than 2 cc., indicating not less than 60 per cent and not more than
80 per cent, by volume, of the acetic-acid soluble fraction of the Oil."
The constituents soluble in the acetic acid solution dissolve,

leaving an oily layer composed chiefly of p-cymene and other
terpene hydrocarbons. Acetic acid solution is used to raise
the lower limit of the oily layer within the graduated portion
of the neck of the flask, because dilution with water would change
the concentration of the acetic" acid and cause some of the
ascaridol to separate out with the oily liquid.
The difference between the volume of the sample used and the
volume of the residual oily layer is considered to be ascaridol.
Thus, a 10 cc. portion of the oil, 3 cc. of which remained undissolved, would contain YLo X 100 = 70 per cent ascaridol.
382

1. Look up the reaction of organic peroxides with acids in a textbook on

organic chemistry and note whether the probable reaction indicated conforms to the information obtained.
2. Why should this assay be considered as only a close appr.oximation?
Assay for Allyl Isothiocyanate.-Volatile oil of mustard is the

only official oil assayed for its content of allyl isothiocyanate.
Black mustard, which is also official, is assayed in a similar
manner, the volatile oil being separated by distillation and its
allyl isothiocyanate content determined.
Exercise 112
Object.-Assay of Oil of Mustard.

Materials Required.-4 cc. of volatile oil of mustard.
100 cc. of alcohol.
50 cc. of 0.1 N AgNO, solution.
5 cc. of 10 per cent ammonia solution.
2 cc. of ferric ammonium indicator solution.
0.1 N NH.SCN solution.
Procedure.-l. "Dilute about 4 cc. of Volatile Oil of Mustard\ accurately
weighed, with sufficient alcohol to make exactly 100 cc. Transfer 5 cc.
of this solution by means of a pipette to a 100 cc. measuring flask, and add
50 cc. of tenth-normal silver nitrate and 5 cc. of ammonia T.S. Connect
the flask to a reflux condenser and heat it on a water bath for one hour."
The reactions which occur may be represented as follows: First,

the ammonia reacts with the allyl isothiocyanate with the formation of thiosinamine:
When the thiosinamine is heated with ammoniacal silver nitrate

solution, it breaks up quantitatively into allyl cyanamide and
hydrogen sulfide:
383
N-C sH5
~C
and the liberated hydrogen sulfide combines with the silver

oxide to form silver sulfide:
2AgNO s + 2NH40H~Ag20
Ag 20 + H2S~Ag2S + H 20
+ 2NH NOs + H 20
4
The complete reaction may, therefore, be represented as follows:

CsH.NSC
+ 3NH s + 2AgNOs~Ag2S +
CNNHCsH.
+ 2NH4NOs
2. "Allow the liquid to cool to room temperature, disconnect the flask

from the condenser, add sufficient distilled water to make the mixture measure 100 cc., mix well, and filter through a filter which has not been previously
moistened. Reject the first 10 cc. of filtrate. To 50 cc. of the subsequent
filt.rate, accurately measured, add about 5 cc. of nitric acid and 2 cc. of ferric
ammonium sulfate T.S., and titrate the excess of silver nitrate with tenthnormal ammonium thiocyanate. Each cubic centimeter of tenth-normal silver nitrate is equivalent to 0.004956 Gm. of allyl isothiocyanate."
Upon filtration, the insoluble silver sulfide is removed, while

the excess of AgNO s passes into the filtrate. This excess of 0.1
N AgNO s is then determined by titration with 0.1 N NH4SCN,
using ferric ammonium sulfate as the indicator. The reactions
which take place are:
AgNO s +
2FeNH 4(S04)2 +
NH4SCN~AgSCN
NH4NOs
6NH4SCN~2Fe(SCN)3
4(NH4)2S04
Each cubic centimeter of 0.1 N AgN0 3 consumed in the reaction with the hydrogen sulfide is equiyalent to 10 X 91~OI;O X 2
0.004947 Gm. allyl isothiocyanate, where 99.13 is the molecular
weight of the latter and 2 is its hydrogen equivalent. In calculating the percentage of aliyl isothiocyanate in the sample, it should
be noted that an aliquot portion of the filtrate corresponding to
one-half of the sample taken is represented in the titration.
384

1. Why is nitric scid used in the titration of the excess AgNO a with
KCNS?
2. Explain each step of the procedure given in the U.S.P. for the sssay
of black mustard.
3. Write equations for all of the reactions that take place in problem 2.
Assay for Volatile Oil inSpirits.-The estimation of the volatile

oil content of the certain official spirits is based upon the separation of the volatile oil by means of an immiscible solvent and
measurement of the volume of the oil as illustrated in the following exercise.
Exercise 113
Object.-Assay of Spirit of Peppermint.

Materials Required.-5 cc. of spirit of peppermint.
A Babcock bottle and centrifuge.
A 1 cc. pipette.
1 cc. of kerosene.
50 cc. of a saturated solution of calcium chloride (about 30 Gm. in sufficient water to make 50 cc.) acidulated with HCl.
Procedure.-l. "Transfer exactly 5 cc. of the Spirtt-to a Babcock bottle,
graduated to 8 per cent, add exactly 1 cc. of kerosene from a pipette calibrated to deliver that amount, and mix well. Then add sufficirnt saturated
calcium chloride solution, acidified with hydrochloric acid l to almost fill the
bulb of the flask. Rotate the flask vigorously to insure
thorough mixing, then add sufficient of the calcium chloride
solution to bring the separated oil into the neck of the
flask."
The acidified calcium chloride solution, when

added to the alcoholic spirit, throws the volatile oil
.~'
out of solution and the oil dissolves in the kerosene
FIG. 64.- when the mixture is shaken. The calcium chloride
Babcock bottIe. (Cour- serves to' "salt out" any volatile oil from the diluted
te8Y
Fi8her alcoholic mixture. The acid insures the absence of
Scientifio
alkali which would render acids, traces of aldehydes,
Company.)
etc., water-soluble. The determination is carried
out in a Babcock bottle (Fig. 64), the neck of which is calibrated
into eight main divisions of 0.2 cc. each.
2. "Centrifuge for five minutes at about 1500 revolutions per minute
and then read the volume of oil in the stem. Subtract 5 divisions for the
385
kerosene added and multiply the remaining number of divisions by 4.2 to

obtain the volume of oil of peppermint in 100 cc. of the Spirit."
The oil-kerosene layer being lighter than the aqueous layer

rises into the neck of the flask when the mixture is centrifuged.
The length of the oil-kerosene layer supplies the information
needed for the calculation of the per cent of oil in the spirit,
e.g., if 7.4 divisions represents the length of the column of oilkerosene in the graduated neck of the flask, the oil measures
7.4 - 5.0 = 2.4 divisions because each 5 divisions are equal to
1 cc. (5 X 0.2 cc.) and .1 cc. of kerosene was added. The 2.4
divisions each of which represent 0.2 cc. are equal to 0.48 CC.
of oil. Since a 5 cc. sample is employed, 100 cc. of the spirit
yielded 0.48 X 20 = 9.60 cc. of oil. The procedure is subject
to slight error which has been experimentally determined.
Consequently, the volume of oil in scale divisions is multiplied
by 4.2 (20 X 0.21 cc.) to correct for the error due to solubility
of the oil-kerosene layer in the aqueous layer and also due
to contraction of volume of the immiscible solvents when mixed.
Thus in the example given, 2.4 X 4.2 = 10.08 cc. of oil in 100 cc.
of the spirit.
Similar assay procedures are applied to a number of the official
spirits in the U. S. Pharmacopoeia as indicated in the following
table:
TABLE LIX.-OFFICIAL SPIRITS ASSAYED FOR VOLATILE OIL CONTENT
Spirit
Anise ......... ..................

Cinnamon ........ .............. .
Lavender ......... " . . ..........
Orange, compound .. ............. .
Peppermint .......................
Spearmint .............. ........ .
Amount
used,
cc.
5
5
10
2
5
~5
Factor
4.2
4.2
2.2
10.5
4.2
4.2
Official
requirement,
per cent
9 to
9 to
4 to
25 to
9 to
9 to
11 V/V
11 V/V
6V/V
30V/V
11 V/V
llV/V
CHAPTER XXIV
ALKALOIDAL ASSAYING ..
Proximate Assays.-Alkaloidal assays are commonly referred
to as proximate assays. When such substances as the alkaloids
and glycosides were first isolated from. vegetable matter, they
were regarded as plant principles which still retained their
vegetable character, which entered immediately into the composition of the plant, and which had not been: altered in composition.
Consequently, they were called proximate principles is opposed
to the ultimate principles, such as water, carbon, acetic acid, and
methyl alcohol which had previously been obtained more ot less
from all plant products by the methods of pyroanalysis (destructive distillation).
General Principles.-The alkaloidal drugs and preparations
derived from them constitute a relatively large1>roportion of the
official substances which are employed frequently in modern
therapy. As a class of medicinal agents, they ar~ ch~racterized
by their high potency. A slight deficiency of alkaloid in a
preparation may cause a marked decrease in physiological
effect; on the other hand, a slight excess may cause toxic effects
when the preparation is administered. It therefore follows that
the accurate estimation of the quantity of alkaloids present in a
medicinal substance is an important subdivision of pharmaceutical analysis.
"The assay of alkaloidal drugs and preparations is generally
performed for purposes of standardization, proof of purity,
commercial evaluation, or pharmaco-Iegal purposes. Methods of
various types have been developed for the quantitative estimation.
of these principles, e.g., gravimetric, volumetric, colorimetric,
potentiometric, and physiological. The official assays are limited
to the gravimetric, volumetric, and physiological methods, only
the first two of which come within the scope of this work.
The amounts of alkaloids which occur in crude drugs are subject
to considerable variation in different samples of the same drug.
386
ALKALOIDAL ASSAYING
387
The variations may be caused by several factors, i.e.: (1) the age
of the plant when it is collected; (2) the season of the year when
the drug is harvested; (3) the soil and climate in which the drug is
grown; (4) the conditions under which the drug is collected,
dried, and stored. The quantity of alkaloid present in galenical
preparations is alscl ,subject to variation due to a number of
factors, some of'--which are: (a) the quality of drug employed;
(b) the menstruum used in the extraction of the alkaloid; (c)
the ltmount of decomposition of the alkaloid during the process
of extraction and during the period of storage.
Many alkaloidal preparations deteriorate comparatively
rapidly, the rate of deterioration being markedly affected by the
nature of the alkaloid, the pH value of the preparation, heat, and
light. Frequent restandardization of the alkaloidal drugs and
their preparations is therefore essential.
In view of the fact that the alkaloids may comprise only a fraction of 1 per cent of the substance assayed and that this small
amount must be separated from numerous other constituents
present in the crude drug or preparation, such as resins, volatile
and fatty oils, coloring matter, glycosides, fatty acids, gums, and
proteins, it is evident tnll1t the exact technique involved in any
given method must be carefully adhered to in order to estimate
the variations in alkaloidal content. It is this special technique
that characterizes the chemicai assay of alkaloidal drugs rather
than the gravimetric or volumetric nature of the procedure
employed.
The principles employed in the quantitative determination of
the alkaloids by chemical methods are based upon certain
characteristic properties of these substances. The following
general properties are possessed by most of the members of this
class of compounds: (1) Alkaloids are usually sparingly soluble
in water but readily soluble in certain organic solvents which are
immiscible with water, such as chloroform, ether, amyl alcohol,
benzene, petroleum benzin, or mixtures of these solvents. (2)
Alkaloids combine directly with acids to form salts which are
usually soluble in water but insoluble in certain organic solvents
such as chloroform and ether. (3) Alkaloids are liberated and
usually precipitated from aqueous solutions of their salts by
alkalies. (4) Alkaloids form highly insoluble precipitates with a
388
considerable number of reagents, especially with the salts of some

of the heavy metals such as mercury, gold, and platinum.
In general, the methods employed to separate the alkaloids
from the other constituents present in a drug involve the use of
the so-called immiscible solvents such as water with ether, water
with chloroform, or water with ether-chloroform mixture. The
process is generally carried out as follows, the separation of the
immiscible solvents being accomplished by means of a separatory
funnel (Fig. 65): An accurately weighed portion of the drug is
treated with a suitable alkali, whereupon the alkaloids,_ which
usually occur in the drug in combination with acids as salts,
are set free. An organic solvent such as ether is
then added, which dissolves the free alkaloids. An
aliquot portion or all of the ethereal solution
corresponding to a definite weight of the sample
taken is separated and shaken successively with
several portions of acidulated water. The complete
extraction of the alkaloids from the ethereal solution
is determined by testing a portion of the latter with
a suitable alkaloidal reagent. The_...alkaloidal salts
formed with the acid dissolve in the water, leaving
most of the resins, coloring matter, fatty SUbstances,
etc., in the ethereal portion. The add extractions
FIG. 65.- obtained are combined, made alkaline, and shaken
Separatory
h
funnel.
with successive portions of et er. The alkaloids
liberated from their salts by the alkali pass into
solution in the ether, leaving water-soluble impurities behind.
The separated ethereal extractives may then be combined,
evaporated to dryness, and weighed; or the residual alkaloid
may be dissolved in an excess of acid of known normality,
and the excess of acid may be determined by titration with a
standard solution of alkali, using a suitable indicator to show
when the end point is reached.
When the above general procedures are applied in assaying,
numerous difficulties may be encountered which will lead to
serious error unless proper precautions are observed, e.g.:
1. The organic solvents employed are sometimes appreciably
soluble in water; thus ether is soluble in twelve times its volume
of water at 25C. Consequently, when an organic solvent
ALKALOIDAL ASSAYING
389
partially soluble in water, such as ether, is employed to extract

the alkaloid from an aqueous solution, some of the alkaloid will
remain in the aqueous solution. This difficulty may be overcome
by extraction with small successive portions of the immiscible
solvent.
2. Resins, fats, and many other organic substances remain in
solution in. the organic solvent if the aqueous liquid employed in
the shaking-out process is acid, but they dissolve in the aqueous
liquid if it is alkaline, forming soaps, resinates, etc. Care should
be exercised, therefore, to have sufficient acid in the aqueous
solution used to react with the alkaloid from the organic solvent
and also to neutralize all of the alkali and leave a slight excess of
acid.
3. When the liquids are shaken in the separatory funnel,
emulsification may occur between the organic solvent and the
aqueous solution, making it exceedingly difficult or impossible to
separate the liquids quantitatively. The formation of emulsions
may frequently be avoided by rotating the separatory funnel
containing the liquids slowly and occasionally inverting it in
place of shaking vigorously. Since the rapidity with which the
alkaloids are extracted from a liquid in which they are insoluble
by an immiscible liquid in which they are soluble is largely
dependent on the rate of change of the intersurface between the
liquids, it is necessary that gentle agitation be continued longer
than is required when strong agitation is employed. If emulsification occurs, it is often possible to break the emulsion by some
one or more of the following procedures: (a) addition of more of
the aqueous or organic solvent; (b) gentle stirring with a glass rod;
(c) drawing off the separated solvent as rapidly as it separates;
(d) addition of a few drops of alcohol; (e) addition of a few cubic
centimeters of salt solution; (f) if alkaline, make add; (g) if acid,
make alkaline; (h) filtration through a pledget of cotton; (i)
drawing off the emulsion and treating it with anhydrous sodium
sulfate. If none of these methods su'ffices to break the emulsion,
it may be necessary to start the assay over. Time is always
saved by taking every precaution to prevent the formation of
emulsions. Emulsions formed in the assay of drugs containing
a large proportion of fatty matter should be treated as
follows:
390
" Add sufficient N sulfuric acid to assure acidity, and evaporate

the volatile solvent, while stirring with a rubber-tipped glass rod.
When the resinous and fatty matter has been agglutinated, cool
the acid solution and filter it through a small, wetted filter into a
separator. Redissolve the residue in 15 cc. of ether, add 5 cc. of
0.1 N acid, evaporate the ether as before, with continued stirring,
and pour the acid solution through the filter into the separator.
Repeat the extraction of the fatty residue with dilute acid two or
three times and finally wash the filter free from alkaloids."
4. When an acid solution contained in a separatory funnel to
which ether has been added is made alkaline, sufficient heat is
frequently generated to volatilize some of the ether and cause
considerable pressure in the funnel. Often the pressure generated
is sufficient to force the stopper out of the funnel with loss
of some of its contents, particularly when the funnel is shaken.
For this reason, it is advisable to permit the mixture to stand and
cool for a short time after adding the alkali and to agitate it
gently at first.
Sources of Error.-Numerous possibilities of error are encountered in alkaloidal assaying other than those_,eommon to all
analytical work, such as those caused by inaccurate weighing,
faulty calibration and reading of measuring apparatus; incorrect
strength of volumetric solutions, dirty apparatus, etc. Some of
the more common errors are caused by: (1) failure to secure
complete extraction of the alkaloids from the drug or from solution; (2) loss of volatile solvent during maceration of the drug
before the aliquot portion is taken; (3) imperfect separation of the
immiscible liquids; (4) decomposition of the alkaloids, especially
those which have an ester structure, upon prolonged contact with
strong acids and alkalies; (5) the use of the wrong indicator.
Other sources of error, such as variation in the moisture content
of the sample, frequently cause different analysts to obtain nonconcordant results in the assay of the same drug. The most
common cause of variations in results obtained by different workers, however, is referable directly to a lack of familiarity with
the procedures involved, since considerable experience in making
assays of this type is essential to acquire proficiency.
Theory of Distribution Coefficient.-According to Nernst's
law, if two practically immiscible solvents are in contact and a
ALKALOIDAL ASSAYING
391
substance which IS'soluble in both liquids is added to them, the

substance will dist,ibute itself between the liquids in such a
way that the ratio OJ the concentrations of the two solutions is a
constant irrespective of the quantity of solute. This constant is
variously termed tfle distribution ratio, distribution constant,
distribution coefficieni, or partition coefficient.
If C1 is used to designate the concentration of an alkaloid in
ether, and C2 to designate the concentration of the same alkaloid
in an equal volume bf an aqueous acid solution, Nernst's law may
be formulated Cda1 = K, where K is the distribution constant
or coefficient. If" ~here are several solutes such as alkaloids,
resins, etc., present in the ether when it is shaken with the acid
solution, the distqbution of each solute is the same as if it were
present alone.
f
Measurable ~E)viations from Nernst's law take place in alkaloidal af!,saying <\he to molccular association and dissociation,
ionization, che:rpical reaction, etc., but the fundamental principle
which it exprpsses may be used to explain why it is better to
extract tM .alkaloid from a solvent with several small portions
instea~ of olle large portion of an immiscible solvent; e.g., assume
that 1 Gmiof atropine in 100 cc. of ether solution when shaken
with an equal volume of dilute sulfuric acid distributes itself so
that O.lrm. remains in the ether layer and 0.9 Gm. in the acid
layer of the solvents, Cacid/Cether = 9 (the distribution coefficient)1 Obviously, only 90 per cent of the atropine present would
be extracted from the ethereal solution by extracting it once with
100 ~c. of dilute sulfuric acid solution. If the extraction is
performed with 200 cc. of the acid solution, the amount of atropine extracted should be x /200 = 9 and x
-x
= 0.947 Gm. atropine.
100
Where x = the amount of atropine extracted from the ether

bt the acid solutio?,
C acid
= 2~~ and
Ce>her
= 11~0 x. If the
original solution is extracted with several small portions of acid

solution, i.e., 50 cc., the amount of alkaloid extracted will be
x
.
1- x
.
x/50
Cacid = 50. and Cether = 100 and smce 1 _ x' == 9, x = 0.818
100
Gm. atropine removed by the first extraction with acid, leaving
392
0.182 Gm. in the ethereal layer.
A second extraction with a
50 cc. portion of acid solution will remove
0.1~15~ Xl
= 9 and
100
= 0.1489 Gm. atropine where Xl equals the amount of atropine
Xl
which passes into the acid solution. Thu~, in two extractions,

0.818 + 0.1489 = 0.9669 Gm. atropine are removed from the
ethereal solution by 100 cc. of acid solution, whereas a single
extraction with ZOO cc. of acid solution removed only 0.947 Gm.
of the alkaloid.
Since economy in the use of solvents is an important consideration and large volumes are difficult to handle in shaking out the
alkaloids, it is more practical to use several small portions of
immiscible solvent than one large portion. In the official assay
processes, 10 to 20 cc. portions of the immiscible solvent are used,
because it has been found that these quantiti,es serve best to
extract the alkaloids quickly and economically.
Choice of Indicators for Alkaloidal Titrations.-In the official
alkaloidal assays by volumetric methods, the use of methyl red or
cochineal indicator solution is recommended ip all of the titrations. The same lot of indicator used in the standardization of
the volumetric solutions should be employed in Ftrating the
alkaloids. Numerous investigators have found that III some cases
other indicators than those employed in the official assays give
more accurate results. The accompanying tabll'l, page 393,
showing the indicators best suited to the titration of some of
the more important alkaloids, is taken with slight modification
from a paper by H. Wales. l
Materials Required.-In the statement of the materials
required under the following respective exercises, it is assumed
that the student's locker is supplied with four separatory funnels
necessary to conduct alkaloidal assays in duplicate, as well as
measuring cylinders, burettes, flasks, etc. It is also assumed
that the following substances are available in the laboratory;
sulfuric acid, hydrochloric acid, ammonia water, distilled water,
alcohol, mercuric potassium iodide test solution, iodine test
solution, and methyl red and cochineal indicator solutions. The
quantities of immiscible organic solvent required are approximate.
1
WALES,
H., Jour. Ind. Eng. Chem., 1926, 18,390.
393
ALKALOIDAL ASSAYING
TABLE LX.-INDICATORS FOR ALKALOIDAL TITRATIONS
Salt of
Average
pH
pH range
for
indicator
Aconitine ................. .
Arecoline ................. .
Atropine .... '" .......... .
Brucine .................. .
Cephaeline. . . . . . . . . . . .. '"
Cinchonine ............... .
Cinchonidine .............. .
COcaine .................. .
Codeine .................. .
Cotarnine ................ .
Diacetyl morphine. .. . .... .
Emetine .................. .
Ethyl hydrocupreine ....... .
Ethyl morphine ........... .
Homa,tropine .............. .
5.04
4.81
5.56
4.85
4.81
6.02
5.90
5.20
4.86
5.97
4.89
4.90
6.33
4.99
5.'14
4.2 to 5.8
3:8 to 5.8
3.8 to'7.2
3.9 to 6.0
4.2 to 5.4
5.5 to 6.5
5.4 to 6.4
4.0 to 6.5
3.6 to 6.3
4.9 to 7.0
4.2 to 5.7
4.2 to 5.6
5.6 to 7.0
4.2 to 5.8
3.9 to 7.6
Hydrastine ............... .
4.45
3.8 to 5.0
Hyoscine ................ "

Hyoscyamine ............. .
4.83
5.83
3.6 to 5.6
3.8 to 7.6
Morphine ................. .
Narcotine ................ .
Nicotijle .................. .
Papaverine ............... .
4.68
4.43
5.26
4.23
4.0 to
3.9 to
4.4 to
3.8 to
Physostigmine ............ .
Pilocarpine ............... .
Quinine .................. .
Quinidine ................. .
Strychnine ................ .
Thebaine ................. .
Yohimbine ................ '1
4.85
4.31
6.12
6.10
4.81
5.08
4.72
3.8 to 6.0
3.6 to 5.0
5.5 to 6.5
5.5 to 6.5
3.8 to 6.0
4.0 to 6.2
4.0to5.3
5.2
4.9
6.1
4.6
Indicator
Methyl red
Methyl red
Methylred
Methyl red
Methyl red
Bromcresol purple
Bromcresol purple
Methyl red
Methyl red
Bromcresol purple
Methyl red
Methyl red
Bromcresol purple
Methyl red
Methyl red, propyl red,
bromcresol purple
No end poin t wi th
methyl red or bromphenol blue
Bromphenol blue
Methyl red, propyl red,
bromcresol purple
Methyl red
Bromphenol blue
Methyl red
Indistinct, cannot be
titrated
Methyl red
Bromphenol blue
Bromcresol purple
Bromcresol purple
Methyl red
Methyl red
Methylred
Alkaloidal Test Solutions.-A number of reagents form insoluble compoubds with many of the alkaloids precipitating the
latter from aqUeous solution. In many cases, the presence of a
mere trace of alkaloid can be detected by adding a suitable
394
QUANTITATIVE PHARMACEUTICAL VHEMISTRY
reagent to its solution. Some of the frequerltly used reagents are

auric chloride, platinic chloride, mercuric chloride, gallotannic
acid, picric acid, phosphomolybdic acid, potassium mercuric
iodide, and iodine in potassium iodide. . While most of these
reagents are employed in qualitative tEljSts for the alkaloids,
only the last two are of importance in the ofl1cial assays, since they
are employed to test for the complete extraction of the alkaloids
by immiscible solvents.
Mercuric potassium iodide test solution, ~ommonly known as
Mayer's reagent, is prepared by dissolving 1.358 Gm. of mercuric
chloride in 60 cc. of distilled water and 5 Gm. of potassium iodide
in 10 cc. of distilled water, mixing the two sblutions and diluting
the product to 100 cc. with distilled watei:. Mayer's reagent
forms curdy yellowish or white precipitates \"ith minute traces
of many of the alkaloids when it is added ip their acidulated
aqueous solutions. The composition of the pr~cipitates usually
varies somewhat depending upon the conditionslunder which the
precipitation is conducted. Mercuric potassium~odide is a very
sensitive reagent for strychnine and quinine; it gives a much less
delicate reaction with atropine, coniine, and .morphine; and colchicine, caffeine, and theobromine are not precipita.ted by it at
all.
\
Iodine test solution, commonly known as Wagner's "re,{(Jent, is a
solution containing approximately 1.2692 Gm. of iodineflnd 1.8
Gm. of potassium iodide dissolved in sufficient distilled "later to
make 100 cc.; 0.1 N iodine solution may be employed. W ~gner's
reagent yields reddish or red-brown precipitates of complex
composition with nearly all of the alkaloids, even in dilute, solutions, when it is added to an aqueous solution slightly acitlified
with sulfuric acid. The precipitates are usually amorphous.
The quantity of the reagent used should not be sufficient to color
the solution more tlian a faint yellow.
General Procedures. 1. Selec,tion of the Sample.-The sam~le
of a crude drug taken for assay must represent the entire drug
and be powdered to a specified fineness. Usually the drug must
be ground to a fine powder (No. 60), but finer powders may be
employed. When the whole crude drug such as leaves or
seeds is powdered for assay, care must be exercised n6t to
exclude leaf stems, seed coats, etc., since the alkaloid may be
ALKAWIDAL ASSAYING
395
localized in amounts varying in the different morphological parts

of the drug. Care should be taken to avoid the loss of water
during the powdering of the drug. If it is impossible to avoid this
loss; the drug should be dried at a low temperature before
powdering, the loss of water noted, and a correction made in the
final calculations.
2. Weighing of Portions for Assay.-In the preparation of
drugs for assay, all portions directed to be weighed should be
weighed accurately, but with crude drugs, accuracy to the second
decimal point is sufficient. Portions of pilular extracts may be
weighed on a piece of waxed or parchmentized paper, the surplus
paper cut away, and the extract and adhering paper dropped into
a separator, beaker, or dish containing the solvent, and the
extract dissolved. In transferring weighed portions to a separator, the dish containing the material to be assayed should be
thoroughly rinsed and the rinsings added to the separator.
3. Methods of Extraction.-The alkaloids are extracted from
drugs by the process of maceration, by the process of percolation,
and by the process of continuous extraction. The maceration
process is adapted to those drugs which contain a relatively
large amount of alkaloids and which, because of their nature or
the nature of the drug, are difficult to extract completely by
the process of percolation. In the maceration process, it is
assumed that all of the alkaloids are dissolved in the organic
solvent and that an aliquot part of the solvent contains an
amount of the extracted alkaloids in definite proportion to
the weight of drug taken as sample. Many analysts believe that
the aliquot portion of solvent obtained by the maceration
process does not contain exactly the assumed proportion of the
total alkaloids. Consequently, methods for the total extraction
of the alkaloids are regarded as more satisfactory.
A. By Maceration.-An accurately weighed portion of the
ground drug is treated with the specified solvent or mixture of
solvents made alkaline with ammonia T.S. and thoroughly
mixed and allowed to macerate for from 12 to 24 hr. with OCCasiona,l agitation, or for a shorter period with continuous agitation.
At the end of this period, the drug is allowed to settle and then
an aliquot portion of the solvent is decanted and used for the
extraction of alkaloids.
396
QUANTITATIVE PHARMACEUTICAL CHEMISTRY;
The organic solvent, accurately measured from a calibrated

cylindrical graduate, is added and allowed to stand in contact
with the powdered drug so that the small particles of the drug
will be impregnated with the organic solvent before the ammonia
is added. If the ammonia test solution is added first, the drug
particles become wetted by the water with which the organic
solvent is immiscible, and the latter will not permeate the drug
cells to extract the alkaloids completely. The alkaloids generally
occur in combination with various acids as salts in the crude drug.
The ammonia reacts with these alkaloidal salts, forming the free
alkaloids and the ammonium salts of the acids, and the liberated
alkaloids dissolve in the organic solvent. The process of extraction of the alkaloid from the cellular structure of the drug is
probably of an osmotic nature which proceeds slowly. Consequently, a period of maceration is required to permit the complete transference of the alkaloids from the drug cells to the
organic solvent. The rate of extraction is accelerated by grinding
the drug to a fine powder, thereby breaking down its cellular
structure, and by frequent agitation of the drug-solvent mixture
so that the solvent in contact with the pa_rticles is changed
frequently. The small pa.rticles of drug which sometimes remain
in suspension in the organic solvent become hydratrd when the
latter is shaken with a little water and settle out. The flask in
which the maceration and agitation of the drug with the solvents are carried out should be tightly stoppered throughout the
entire period of maceration to prevent the escape of volatile
organic solvent, since the volatilization of some of the solvent
might cause a considerable error; e.g., if 100 cc. of solvent is taken
for the extraction of the alkaloids from a drug and 10 cc. is
lost by volatilization, and an aliquot portion of 50 cc. representing
one-half of the weight of the sample is measured for assay, the
aliquot portion will Mntain five-ninths instead of one-half of the
alkaloids contained in the drug, and an error of approximately
5 per cent will be caused in the assay result.
An aliquot portion of the supernatant organic solvent is
poured off from the mixture and measured accurately in a 9alibrated cylinder; e.g., if 4 Gm. of a drug is macerated with 100 cc.
of solvent and a 50 cc. portion of the solvent is decanted, the
50 cc. portion of solvent will contain an amount of extractive
ALKALOIDAL ASSAYING
397
equivalent to 59100 X 4 = 2 Gm. of the drug, provided that none

of the solvent originall;v added is lost by volatilization. The
aliquot portion of solvent should be measured at the same temperature at which the original solvent was measured, since the
volume of liquids varies somewhat with changes in temperature.
B. By Percolation.-An accurately weighed quantity of the
ground drug is placed in a suitable container and completely
saturated with the specified solvent or mixture of solvents and
allowed to stand for 5 min. A sufficient quantity of ammonia
T.S. to make the drug alkaline is added and thoroughly mixed
with the drug. The moistened drug is transferred to a cylindrical
percolator, previously prepared by packing the outlet with
purified cotton. A small amount of the solvent may be used to
rinse the container and the rinsing added to the percolator.
The drug is allowed to macerate for a suitable period of time (from
1 to 12 hr. or over night, depending upon the drug to be assayed),
then the drug is firmly packed, a pledget of purified cotton placed
above it, and percolated slowly until the drug is completely
extracted. Complete extraction of the alkaloid is determined by
evaporating about 4 cc. of the last percolate to dryness, dissolving
the residue in dilute acid, and adding a drop of mercuric potassium iodide T.S. or, when testing for caffeine or colchicine, a drop
of iodine T.S. There should be no turbidity produced by these
reagents. The percolate is then treated for the extraction of the
alkaloids.
The extraction of the alkaloids from a drug by cold percolation
has the disadvantage that it requires a relatively large amount
of solvent to effect complete extraction. The cold. percolation
process of extraction is best suited to the extraction of drugs
which contain unstable alkaloids or which yield a large amount
of non-alkaloidal extractive by hot percolation. 'The cold
percolation process is well suited to the extraction of those drugs
which contain a comparatively small percentage of alkaloids
and which can be readily extracted by percolation. The importance of the total extraction of the alkaloid from a drug such as
hyoscyamus becomes apparent when it is recalled that the official
drug is required to contain only 0.040 per cent of alkaloids.
The percolator employed in the extraction of the drug should
be of a special type similar to that illustrated in Fig. 66. The
398
barrel of the percolator should be about 15 or 16 cm. long and

about 3 or 4 cm. in diameter. A percolator of these dimensions
will accommodate from 10 to 25 Gm. of drug, and, because of
the small diameter, a long column of the drug will be provided
through which the menstruum must pass. The glass stopcock
is necessary to eliminate the use of rubber connections, since
rubber is dissolved to some extent by chloroform and ether.
The alkaloids are liberated by the ammonia and they dissolve
in the organic solvent as described under A.
f--05
-~ ~
C. By Continuous Extraction.-An accurately
i
weighed portion of the ground drug is placed in
i
an extraction thimble and the thimble transferred to a suitable extractor (a Soxhlet extractor
of appropriate size is satisfactory). The drug is
:
I
!
i
moistened with the specified solvent and mixed
2:
by means of a stirring rod and allowed to stand
:
~1
about 5 min. It is then made alkaline with
~18~
~ the specified quantity of ammonia T.S. and
-'--. \. ,I.~" thoroughly mixed.
The stirring rod is rinsed
~ .7;. 10
with a small portion of th~ solvent and the drug
- r... :.
macerated for from 6 to 12 hr. or over night.
The drug is covered wi~h a pledget of purified
FIG. 66.-Per- cotton and packed in the thimble, a sufficient
cola tor
recom- quantity of solvent is added and the drug
mended for type
process B (meas- extracted for a specified period of time. The
urements are given solvent remaining in the extraction chamber is
in millimeters).
transferred to the receiving flask and the liquid
extract treated for the extraction of alkaloids.
An extraction apparatus of the type illustrated in Fig. 60
and described on page 337 is very satisfactory. The continuous
extraction process requires a small amount of the organic solvent
and, in most cases; complete extraction of the alkaloids is obtained
in from 3 to 5 hr.
4. Extraction of the Alkaloids from Organic Solvents with Acid.The alkaloids are extracted from ethtlreal or chloroformic
solutions by shaking with dilute acids. The concentration and
quantity of the acid employed are left to the discretion of the
analyst, but the use of 10 cc. of a normal or 5 per cent acid or
sufficient to render the mixture distinctly acid for the first extrac-
1-
I
oil
~
1
!
Le.
ALKALOIDAL ASSAYING
399
tion and of 10 cc. portions of 0.5 N or 2 per cent acid for succeeding extractions will be found to be practical and convenient. The
shaking of the immiscible solvents should not be violent, but it
should be sufficient to mix them thoroughly without emulsification. Repeated successive extractions with small portions of the
acid must be continued until the alkaloid is completely removed
from the ethereal or chloroformic liquid as shown by tests with
the designated reagents. Each portion of the acid solution
should be carefully separated from the organic solvent and run
directly into another separatoryfunnel. In all assays, the extraction should be continued until 0.5 cc. of the last acid washing
shows no turbidity on the addition of a drop of mercuric
potassium iodide T.S., or, in the case of caffeine and colchicine,
on the addition of a drop of iodine T.S. If traces of organic
solvent are drawn off with the acid solution, they may be
removed by washing the combined acid extractives with 2 to 3
cc. of ether or chloroform which is separated and washed in turn
with about 5 cc. of distilled water, the aqueous washings being
separated and added to the acid solution.
If sulfuric acid is used, the acid converts the free alkaloids dissolved in the organic solvent into their sulfates which pass into
solution in the aqueous acid layer of the immiscible solvents when
the mixture is shaken, leaving chlorophyll, resins, fixed and
volatile oils, and other impuritIes dissolved in the organic solvent.
The rate at which the alkaloids pass from solution in the organic
solvent into the acid solution is increased greatly by shaking the
mixture, since a greater area of contact surface between the
liquids is produced. Sulfuric acid solution is employed to extract
the alkaloids, because the ammonium sulfate formed in the next
step in the procedure is not appreciably solub~e in the organic
solvent used for the final extraction of the alkaloid.
5. Extraction of the Alkaloids from Acid Solution with Organic
Solvents.-The acid solution of the alkaloids contained in a
separatory funnel is made alkaline, in most cases with ammonia
T.S., and then shaken with several successive portions of organic
solvent, such as ether, chloroform or ether-chloroform mixture.
The volume of organic solvent used in each extraction should be
at least one-half that of the aque<;ms solution. The extraction
with portions of organic solvent must be repeated as long as any
400
alkaloid is extracted by the solvent. To determine the completion of extraction, evaporate 1 cc. of the last washing and dissolve
the residue in a few drops of diluted hydrochloric acid; the resulting solution should show no turbidity on the addition of a drop
of mercuric potassium iodide T.S., or, in the case of caffeine and
colchicine, on the addition of a drop of iodine T.S. The number
of extractions required depends largely on the character of the
alkaloid. With most alkaloids it is advisable to extract three or
four times before testing. Physostigmine and pilocarpine
require about twice as many extractions as other alkaloids.
The ammonia reacts with the alkaloidal sulfates contained in
the aqueous solution to form the free alkaloids and ammonium
sulfate. If considerable quantities of alkaloid are contained in
the acid solution, the alkaloids may be precipitated when the
alkali is added. Consequently, the first portion of organic
solvent is added prior to the addition of the ammonia water to
dissolve any alkaloidal precipitate formed. If this procedure is
not followed, the precipitate frequently agglomerates into large
particles which dissolve slowly when the organic solvent is
added.
6. Evaporation of Organic Solvents.-The evaporation of the
final alkaloidal solution in organic solvents may be perfllrmed by
distillation with recovery of the solvent, by evaporation on a
steam bath or sand bath, or in a current of warm air. When the
evaporation is conducted on a steam bath, precautions must be
observed to prevent the ignition of the vaporized solvent. The
operation may be performed by heating a prepared water bath
until the water boils, turning out the flame, and placing the
alkaloidal solution contained in a suitable container on the bath.
If a sand bath is employed, it should preferably be of the steamheated type and the temperature of the bath should be about
60C.
Chloroform tends to form unstable compounds with many
alkaloids; consequently, when chloroformic solutions of the
alkaloids are evaporated, the residue may retain traces of
chloroform rather tenaciously. These traces of chloroform are
best removed by treating the alkaloidal residue with 2 to 3 cc.
of ether or alcohol and evaporating the solution to dryness again.
This is a wise pr~caution to employ in the evaporation of all
ALKALOIDAL ASSAYING
401
solutions of alkaloids in organic solvents when the final determination is made by volumetric methods in order to remove small
traces of ammonia which may be occluded in the alkaloidal residue, particularly if the latter is gummy in nature.
Those alkaloids which have the structure of esters such as
atropine, hyoscyamine, and cocaine and some others are decomposed more or. less by heat and by prolonged contact with water
and strong acids and alkalies. Consequently, their solutions
should be evaporated at a low temperature, usually at about
60C.; and, if it is necessary to allow the alkaloid in solution to
stand from one laboratory period to the next before the assay can
be completed, it should be shaken out of acid or alkali solutions
into an organic solvent and kept in a tightly stoppered flask.
7. Gravimetric Determinations of Alkaloids.-If the alkaloid is to
be estimated gravimetrically, the solvent is evaporated in the
usual manner from a previously weighed evaporating dish,
wide-mouth flask, or beaker, arid the residue is dried to constant
weight in an electric or hot-air oven and weighed. The same
procedure should be followed in drying the flask or beaker used
as is employed in drying the container with its residue. When
the residue is to be weighed, if the final solvent has been chloroform, the last traces of that solvent should be removed by
the addition of a little neutral ether or alcohol, followed by
evaporation. Care must be taken to avoid loss by decrepitation,
especially when evaporating chloroformic solutions of nux
vomica or cinchona alkaloids. Decrepitation may usually be
prevented by the addition of a little alcohol after the solution
has been reduced to a volume of 1 or 2 cc., evaporating at a low
temperature, and rotating the container during the process.
8. Volumetric Determinations of Alkaloids.-If tlie alkaloid is
directed to be assayed volumetrically, the residue should be
softened by the addition of about 1 cc. of neutral alcohol or ether,
the required amount of standard acid added, and the mixture
gently warmed to insure the complt'~te solution of the alkaloid.
If preferred, the alkaloidal residue may be dissolved in chloroform, the standard acid added, and the chloroform removed by
evaporation. Before titrating add a sufficient quantity of distilled water to make the volume of the mixture measure about
25 cc. The amount of acid in excess of that which combines
402
.QUANTITATIVE PHARMACEUTICAL CHEMISTRY
with the alkaloid is then determined at once by titration with

standard alkali.
As a rule the volumetric method of determining the quantity
of alkaloid obtained in a given assay is more accurate than the
gravimetric method, since many impurities which cannot be
removed are present in the alkaloidal residue and are computed as
alkaloid when the residue is weighed. If the residue is titrated,
however, only those impurities which are acidic or basic affect
the result.
Adsorbents.-In assaying fiuidextracts, tinctures, and other
preparations of alkaloid-bearing drugs, it is often necessary to
evaporate these to dryness and, to avoid loss and to aid in the
evaporation, they are usually added to some adsorbent material.
Paper pulp or asbestos fiber should be used for this purpose.
Such adsorbent material must be acid and alkali washed and then
rendered neutral by washing with distilled water and dried before
being used.
Apparatus for Proximate Assays.-When a container of definite
size and shape is recommended in a proximate assay process, it
is understood that this is advisory and not ,Poligatory, except
when volumetric flasks, measuring burettes; or other exact
measuring apparatus are specified.
CHAPTER XXV
OFFICIAL TYPE METHODS
The official alkaloidal drugs and the galenical preparations
containing alkaloids are assayed by two type methods and
by special methods. The type methods are (1) the aliquot part
method for drugs and galenical preparations and (2) the total
extraction method for drugs and galenical preparations. The
procedures in the following assays are taken almost verbatim from
the U.S.P. and N.F., slight changes being introduced where
necessary to include more specific details.
Gravimetric Assays by the Aliquot-part Method
Exercise 114
Object.-Assay of Hydrastis for Ether-soluble Alkaloids.

Materials Required.-10 Gm. of hydrastis in No. 60 powder.
200 cc. of ether.
Procedure.-l. "Place 10 Gm. of Hydrastis, in fine powder and accurately
weighed, into a suitable container, add 100 cc. of ether, allow the mixture to
stand about five minutes, and add 10 cc. of ammonia water. Shake the
mixture fOT one hour in a mechanical shaker or during two hours intermittently, and then set it aside overnight. Again shake it during one half hour,
allow the drug to settle, and decant 50 cc. of the clear ethereal solution."
(See General Procedure 3A, page 395.)
The alkaloids found in the rhizomes and roots (If hydrastis consist chiefly of hydrastine and berberine in varying proportion.
The latter is almost inactive, while the hydrastine is very active
physiologically. Consequently, a determination of the total
alkaloids of hydrastis would not ~be a correct measure of the
therapeutic activity of the drug. Ether is used, therefore, as the
organic solvent in the extraction of the alkaloids, because hydrastine is readily soluble and berberine is practically insoluble in this
solvent. The aliquot portion of 50 cc. of the ethereal solution is
equivalent to 59100 X weight in grams of the sample of hydrastis.
403
404
2. "Extract the alkaloids completely from the ethereal solution by shaking with successive small portions of dilute sulfuric acid (about 1 per cent).
Combine the acid solutions in a separator, add about an equal volume of
ether, then add a slight excess of ammonia water, and immediately shake to
extract the alkaloids." (See General Procedure 4, page 398.)
The sulfuric acid converts the hydrastine into hydrastine

sulfate which is soluble in the acid solution.
3. "Separate the ethereal layer, and complete the extraction of the
alkaloids from the alkaline aqueous layer by shaking with successive portions
of ether." (See General Procedure 5, page 39,9.)
The hydrastine liberated from its sulfate is practically insoluble

in water, but it dissolves in ether to the extent of 1 Gm. in
175 cc. at 25C. Traces of berberine carried over into the acid
solution are eliminated by using ether as the immiscible solvent,
since berberine is soluble in water (1 Gm. in 4.5 cc. at 22C.)
and insoluble in ether.
4. "Filter the ethereal solutions and evaporate the ether. Dry the residue to constant weight at 100C.
"The weight obtained represents the yield of anhydrous ether-soluble
alkaloids from 5 Gm. of Hydrastis." (See General "procedures 6 and 7,
page 400.)
The percentage of ether-soluble alkaloids contained in the root

is found by simple calculation as follows:
wt. of alkaloidal residue X 100 t
0.5 X wt. of hydrastis
- per cen .
The Pharmacopoeia requires that hydrastis contain not less
than 2.5 per cent of ether-soluble alkaloids. Compare the percentage of alkaloids found in the sample assayed with the official
requirement.
The extract, fluidextract, and tincture of hydrastis are assayed
by procedures 'similar to that employed in the assay of hydrastis.
1. Why is hydrastis assayed for its content of ether-soluble alkaloids?

2. Explain in detail how to test fOf hydrastine in the ethereal solution
which has been shaken out with acid.
3. Can hydrastine be estimated volumetrically (see table of indicators,
page 286)?
4. What is an aliquot portion of a solution?
405
5. Why is it more practical to use a number of small portions of immiscible

solvent in shaking out the alkaloids than to use one or two large portions?
Exercise 115
Object.-Assay of Cinchona for Total Alkaloids.

Materials Required.-5 Gm. of cincona in No. 60 powder.
200 cc. of ether-chloroform mixture.
Procedure.-l. "Place 5 Gm. of Cinchona, in fine powder, and 15 cc. of
3 per cent hydrochloric acid in a 500-cc. flask and heat the mixture on a
water bath for one hour. Cool and add 200 cc. of ether-chloroformic solution (ether, 3 volumes, chloroform, 1 volume) and 10 cc. of stronger ammonia
T.S. Stopper the flask tightly and shake it for one hour in a mechanical
shaker. Allow the mixture to stand over night, again shake it for one-half
hour, and then allow the drug to settle. (If the supernatant liquid is not
clear, add a few cc. of distilled water, again shake the contents of the flask
vigorously, and allow the drug to settle.)" (See General Procedure 3A,
page 395.)
Cinchona bark contains about 20 alkaloids the most important

of which are quinine, quinidine, cinchonine, and cinchonidine.
The alkaloids occur as salts in combination with quinic and cinchotannic acids. These salts are decomposed when boiled with
hydrochloric acid with the formation of the corresponding
alkaloidal hydrochlorides which are soluble in the water. When
the aqueous solution is made alkaline with ammonia, the liberated
alkaloids pass into solution in the ether-chloroform mixture
almost completely, due to the large volume of the latter
employed.
2. "Quickly decant 160 cc. of the clear, ether-chloroformic solution, measured at approximately the same temperature as the original ether=chloroformic solution and representing 4 Gm. of drug." (See General Procedure 3,
page 395.)
The aliquot portion is equivalent to

the sample taken.
16%00
X the weight of
3. "Transfer the solution to a separator, rinse the measuring vessel with

a small quantity of the original menstruum and add the rinsings to the
separator. Completely extract the alkaloids with approximately 5 per cent
sulfuric acid and collect the acid solution of tIle alkaloids'in a second separator." (See General Procedure 4, page 398.)
The alkaloidal sulfates are formed and dissolve in the acid

solution.
406
4. "Make the acid solution strongly alkaline with ammonia T.S. and
completely extract the alkaloids with chloroform. Evaporate or distil the
chloroform in a tared beaker or flask and dry the alkaloidal residue to constant weight at 100C. The weight obtained, multiplied by 25, represents
the per cent of the alkaloids of cinchona in the drug." (See General Procedures 5, 6, and 7, page 399.)
The ammonia water reacts with the alkaloidal sulfates,

setting the alkaloids free and forming ammonium sulfate.
Only a slight excess of ammonia should be used to make t~e
solution alkaline, since the cinchona alkaloids are appreciably
soluble in strong ammonia solution. The alkaloids set free by
the alkali dissolve in the chloroforI,n.
It has been reported that the cinchona alkaloids form addition
products with chloroform which are not entirely broken down
when the residue from the chloroformic solution of the alkaloids
is dried at 100C. The error in the assay results caused by the
retention of chloroform by the alkaloidal residue when the latter
is dried as directed, however, is negligible.
The gravimetric method is employed in the determination
of the alkaloids in the residue, because the indicator end point
is not sharp when the mixture of alkaloids is titrated.
The U.S.P. requires that cinchona contain not, less than 5 per
cent total alkaloids when assayed by the above method. Calculate the percentage of alkaloids found in the sample assayed
and compare the results with the official requirement.
1. Why is hydrochloric acid used in the initial menstruum to extract the
alkaloids from cinchona?
2. Does the ether-chloroform mixture extract practically all of the alkaloid
present in the sample in the first step?
3. Why should the addition of a large excess of ammonia to the acid
solution of the alkaJoids previous to the shaking out with chloroform be
avoided?
Exercise 116
Object.-Assay of Compound Tincture of Cinchona.

Materials Required.-50 cc. of compound tincture of cinchona.
200 cc. of ether.
Procedure.-l. "Accurately measure 50 cc. of Compound Tincture of
Cinchona and evaporate it, at a temperature not exceeding 100C., to a
407
volume of about 10 cc. Add sufficient asbestos fiber or paper pulp to

adsorb the liquid, and continue the evaporation to dryness. Transfer the
residue to a flask or bottle, add 200 cc., accurately measured, at room temperature, of ether-chloroform mixture (ether 4 volumes, chloroform 1 volume) and sufficient ammonia T.S. (which may be used to rinse out the
adhering portions of the Tincture from the evaporating dish) to render the
mixture strongly alkaline. Securely stopper the container and shake it
mechanically during one hour, or intermittently during two hours, and then
allow the mixt,ure to stand over night."
The tincture is evaporated to drive off the alcohol which is

miscible with ether and chloroform. An adsorbent such as
asbestos fiber or paper pulp (filter paper torn into small pieces)
is added so that the non-volatile residual matter consisting
largely of alkaloidal hydrochlorides, glycerin, and cinchona
red will be deposited as a thin film when the tincture is evaporated. If some adsorbent is not used, a resinous, varnish-like
residue usually results from which the alkaloids cannot be
extracted readily. The addition of ammonia T.S. liberates the
alkaloids which dissolve in the ether-chloroform mixture along
with the glycerin, cinchona red, and considerable other extractive
matter.
2. "Again shake the mixture intermittently for half an hour, allow it to
settle, quickly decant 160 cc. (representing 40 cc. of the Tincture) of the
approximately clear liquid. Filter this into a separator and wash the
measuring vessel with sufficient of the ether-chloroform mixture, adding
the rinsings to the filter. Extract the alkaloids from the clear liquid with
acidulated water, using sufficient dilute sulfuric acid to render the contents
of the separator and each extract distinctly acid to litmus paper. Pass the
acid extracts in succession through a wetted, double filter into a second
separator. Render the combined liquids distinctly alkaline ,with stronger
ammonia T.S., and extract with chloroform. Pass the chloroformic
extracts through a double filter, which is kept saturated with chloroform,
into a suitable, tared receptacle. Evaporate the chloroform on a water
bath, dry the residue to constant weight at 100C., and weigh. The weight
multiplied by 2.5 indicates the weight of alkaloids in 100 cc. of the Compound Tincture of Cinchona." (See Assay of Cinchona, Procedures 2, 3,
and 4, page 405 for explanation.) "
Volumetric Assays by the Aliquot-part Method

Exercise 117
Object.-Assay of Ipecac for Ether-soluble Alkaloids.

Materials Required.-lO Gm. of ipecac in No. 60 powder.
200 cc. of ether.
408
Procedure.-l. "Place 10 Gm. of Ipecac in fine powder in a dry, 250-ec.

flask. Add 100 ee. of ether, which is free from peroxide, stopper the flask,
shake the mixture thoroughly, and allow it to stand for five minutes; then
add 10 ee. of ammonia T.S. Again stopper the flask tightly and shake it for
one hour in a mechanical shaker, or intermittently during two hours.
Allow the mixture to stand over night, again shake it intermittently during
one-half hour and then allow the drug to settle. Decant into a separator
50 cc., representing 5 Gm. of Ipecac, aocurately measured, of the clear,
supernatant liquid, rinsing the measuring vessel with a small. quantity of
ether and adding the rinsings to the solution in the separator." (See General Procedure 3A, page 395.)
Ipecac contains a number of alkaloids, the most important of

which are emetine, cephaeline, and psychotrine. These three
alkaloids occur in varying proportions in different varieties of
the root. Psychotrine constitutes a relatively small proportion
of the total alkaloids and is not known to be of importance
therapeutically. Consequently, this alkaloid is eliminated in
the assay process, since it is practically insoluble in ether, while
the liberated emetine and cephaeline, being soluble in ether,
are determined. The ipecac must be finely powdered, since it
is not penetrated readily by the solvent due to its high starch
content.
~
The aliquot portion of the ethereal solution cOIftains an
amount of emetine and cephaeline equivalent to one-half the
weight of sample taken.
2. "Completely extract the alkaloids from this ethereal solution with
approximately normal sulfuric acid, preferably using 15 cc. the first time
and 10 cc. on each succeeding extraction, and filtering each portion into a
second separator. Continue the extraction until no reaction can be detected
in the sulfuric acid solution when tested." (See General Procedure 4,
page 398.)
The sulfates of emetine and cephaeline are formed and dissolve

in the aqueous solution.
3. "To the combined acid solutions add about an equal volume of peroxide-free ether, render the mixture alkaline by the addition of ammonia ~
T.S., and extract with successive portions of the ether until nq visible reaction takes place when tested as directed above. Filter each portion of the
ethereal extract int~ a 200-cc. flask or beaker, and carefully evaporate the
combined ethere~l solutions on a steam bath until nearly but not quite dry.
Add 5 cc. of the peroxide-free ether and again evaporate nearly to dryness.
Add 10 cc. of tenth-normal sulfuric acid and heat on a steam bath to effect
409
complete solution and to remove all of the ether. Cool, and titrate the
excess of acid with tenth-normal sodium hydroxide, using methyl red T.S.
as the indicator. Each cubic centimeter of tenth-normal sulfuric acid is
equivalent to 0.0240 Gm. of the ether-soluble alkaloids of Ipecac." (See
General Procedures 5, 6, !ltnd 7, page 399.)
The ammonia sets the emetine and cephaeline free, and the
liberated alkaloids dissolve in the ether.
The residue consists almost entirely of emetine and cephaeline
which dissolve in the standard acid solution, forming emetine
and cephaeline sulfates, as illustrated by the following reaction
for emetine:
2C16H2202N
+ H2S0C~(C15H2202N)2.H2S04
Each cubic centimeter of 0.1 N H 2 S0 4 consumed corresponds to

0.024 Gm. of the ether-soluble alkaloids of ipecac.
The Pharmacopoeia requires that ipecac contain not less than
1.75 per cent of ether-soluble alkaloids. Calculate the percentage
of ether-soluble alkaloids contained, in the sample assayed and
compare the results with the official requirement.
Fluidextract of Ipecac, U.S.P., and Tincture of Ipecac, N.F., are
assayed for total ether-soluble alkaloids by procedures which
involve the same principles as the assay of ipecac.
1. Why is ipecac assayed for its content of ether-soluble alkaloids?
2. Ascertain the molecular weights of emetine and cephaeline from the
table, U.S.P. page 587, and show how the alkaloidal equivalent of each
cubic centimeter of 0.1 N H 2S0 4 is derived.
8. Would the error in the assay result be very considerable if the ethersoluble alkaloids of the ipecac consisted almost entirely of emetine?
4. Enumerate several possible sources of error which might be encountered
in the assay of ipecac.
Exercise 118
Object.-Assay of Areca.
Materials Required.-15 Gm. of a'teca.
250 cc. of ether.
15 of a saturated solution of sodium bicarbonate (about 1 in 10).
About 3 Gm. of sodium bicarbonate.
Procedure.-l. "Place 15 Gm. of Areca, in moderately fine powde!' and
accurately weighed, into a suitable flask, add 150 cc. of ether, allow to stand
about five minutes, then add 15 ce. of a saturated solution of sodium bicar-
410
bonate. Agitate the mixture in a mechanical shaker for one hour, or intermittently during two hours, set it aside overnight, and again agitate in a
mechanical shaker for one hour. Separate 100 cc. of the clear ethereal
liquid." (See General Procedure 3A, page 395.)
The most important alkaloid occurring in areca is arecoline.

A number of other alkaloids, namely, arecaidine, ethyl arecaidine,
guvacine, iso-guvacine, guvacoline, and arecolidine, have also
been found to be present in the drug in small amounts. The
alkaloids which are largely combined as salts in the drug are
liberated by the sodium bicarbonate. The arecoline and
guvacoline dissolve in the organic solvent while a large proportion
of the other alkaloids, e.g., guvacine and arecaidine, remain in
the aqueous alkaline liquid because they contain carboxy groups
and form water-soluble salts with alkalies. Since arecoline is
the methyl ester of arecaidine and guvacoline is the methyl ester
of guvacine, the carboxy groups are blocked, and these alkaloids
do not form salts with alkalies.
2. "Extract the alkaloid completely from the ethereal solution with successive portions of dilute sulfuric acid (about 1 per cent). Render the
combined acid solutions alkaline with sodium bicarbonate, cautiously
added, to avoid loss by effervescence." (See General Procedure 4, page
398.)
The arecoline and guvacoline which are soluble in \ all proportions in water and in ether form salts of the acid which are soluble
in water but very slightly soluble in ether.
3. "Extract the alkaloid completely from the alkaline liquid with successive portions of ether. Filter the ethereal solutions successively into 10 cc.
of tenth-normal sulfuric acid, rotating after each addition. Evaporate the
ether from this mixture, and determine the alkaloid by titration of the
excess acid with fiftieth-normal sodium hydroxide, using cochineal T.S. or
methyl red T.S. as the indicator.
"Each cubic centimet!-lr of tenth-normal sulfuric acid represents 0.0155
Gm. of arecoline, CSH1302N." (See General Procedures 5, 6, and 7,
page 399.)
The free alkaloids dissolve in the ether. When the ethereal

solution is added to the acid and evaporated, the alkaloids form
salts with a part of the acid and the amount of acid in excess is
determined by titration.
2CsH 130 2N
+ H2S04-(CsH1302N)2.H2S04
411
The National Formulary requires that areca contain not less

than 0.2 per cent of alkaloids calculated as arecoline. Calculate
the per cent of alkaloids contained in the sample assayed.
TABLE LXI.-DRUGS AND PREPARATIONS ASSAYED BY THE ALIQUOT-PART
METHOD
Substance
Amount
used,
Gm.
or cc.
Assayed
for
Official
requirement,
per cent
U.S.P.
Cinchona ...............
Cinchona, compound tincture of ...............
Ipecac .................
Total alkaloids
(n.l.t.) 5
50
10
0.4 to O.5W jV
(n.Lt.) 2
Ipecac, fluidextract of. ...
10
Total alkaloids
Ether-soluble
alkaloids
Ether-soluble
alkaloids
N.F.
Areca ..................
Hy\lrastis ...............
15
10
Arecoline
Ether-soluble
alkaloids
Ether-soluble
alkaloids
Ether-soluble
alkaloids
Ether-soluble
alkaloids
Ether-soluble
alkaloids
(n.l.t.) 0.2
(n.l.t.) 2.5
Hydrastis, extract of .....
2*
Hydrastis, fluidextract of
5*
Hydrastis, tincture of ....
25*
Ipecac, tincture of .......
50*
1. 8 to 2. 2W jV
to 11
2.25 to 2. 75W jV
0.45 to 0.55WjV
0.18 to 0.22W jV
(n.Lt.) = not less than.

* No aliquot part taken.
Alkaloidal Assays by the Total-extraction Method

Exercise 119
Object.-Assay of Hyoscyamus for Total Alkaloids.

Materials Required.-25 Gm. of hyoscyamus in No. 60 powder.
10 cc. of alcohol.
250 cc. of ether.
Procedure.-l. "Place 25 Gm. of Hyoscyamus, in fine powder, in an
extraction thimble, insert the thimble in a Soxhlet extractor, moisten the
412
drug with a mixture of 8 cc. of stronger ammonia T.S., 10 cc. of alcohol and
20 cc. of ether, and mix thoroughly. Macerate the mixture over night, then
extract it for not less than three hours on a water bath, using 'ether as the
solvent. The following alternative process may be used: Moisten 25 Gm.
of Hyoscyamus, in fine powder, with a mixture of 8 cc. of stronger ammonia
T.S., 20 cc. of ether and 10 cc. of chloroform, in a small Ilercolator, previously
prepared by packing the outlet with a pledget of purified cotton. Macerate
the mixture over night, paek it in the percolator and extract by slowly percolating with a mixture of 3 parts of ether and 1 part of chloroform, by
volume. Continue the percolation until the 3 or 4 cc. of percolate last
passed, when evaporated to dryness and the residue dissolved in dilute
sulfuric acid, fails to become turbid when treated with mercuric potassium
iodide T.S." (See General Procedure 3B and C, page 397.)
Hyoscyamine, the chief alkaloid of hyoscyamus, and small

quantities of other alkaloids' are set free from the acids, with
which they are combined in the crude drug by the ammonia, and
dissolve in the organic solvent. The laevorotatory hyoscyamine
is largely converted into its racemic isomer, atropine, in the
ammoniacal solution.
2. "Evaporate the extractive, obtained by either method, to about 15 ce.,
then add 10 cc. of approximately tenth-normal sulfuric acid and 10 cc. of
distilled water, and continue the evaporation until the -:Olatile solvents are
removed. Filter this mixture, collecting the filtrate in a separato:r, dissolve
the chlorophyll residue in chloroform, add acidulated water, 'evaporate on a
water bath until thc chloroform is removed, and filter into the same separator
through the filter previously used."
The large volume of liquid obtained, especially by cold percolation, is concentrated to a small volume to secure a convenient
amount. It should not be evaporated to dryness because the
chlorophyll and other non-alkaloidal residue make it difficult
to completely dissolve the alkaloids in the acid. The alkaloids,
hyoscamine and atropine, being esters undergo hydrolysis readily
in aqueous acid and alkaline solution especially when heated.
Consequently, the evaporation of the organic solvent should be
conducted at a low temperature, preferably not exceeding 40C.,
after the addition of the acid. The evaporation may be carried
out advantageously in a vacuum desiccator under reduced pressure. Upon filtration, the non-alkaloidal residue consisting
mostly of chloropyll is retained on the filter. This residue is
dissolved in chloroform, treated with acid, and evaporated to
413
completely extract the alkaloids. This method of obtaining

the alkaloids in acid solution from the organic solvent avoids
the formation of the stable emulsions frequently obtained by the
shaking out process. -Quite stable emulsions are particularly
liable to be encountered in shaking out alkaloids in the assay of
hyoscyamus, belladonna, and stramonium owing to the' presence
of small amounts of saponins which act as emulsifying agents.
The sulfates of the alkaloids are formed and dissolved in the
acid solution leaving most of the coloring matter! etc., in the acid
insoluble residue
3. "Render the mixed filtrates alkaline with ammonia T.S. and remove
the alkaloids by extracting with chloroform, testing for the complete extraction of the alkaloids." (See General Procedure 5, page 399.)
The alkaloids are set free by the ammonia water and dissolve
in the chloroform.
4. "Evaporate or distil the chloroform from the combined extractions
until reduced to a small volume, then evaporate to dryness on a water bath,
and keep at this temperature for fifteen minutes. Dissolve the residue in
chloroform, evaporate to dryness on a water bath, and continue the heating
for fifteen minutes. Repeat this treatment for the third time."
The solution of the alkaloids in organic solvent is evaporated

to drive off the solvent and ammonia. The residue is repeatedly
dissolved, evaporated, and dri,ed to drive off small amounts of
volatile amines which are basic and, unless removed, would
neutralize some of the acid in the next step of the procedure with
subsequent high results.
5. "Dissolve the resulting residue in ch\oroform, add 15 cc. of fiftiethnormal sulfuric acid, remove the chloroform by evaporation, and titrate the
excess acid with fiftieth-normal sodium hydroxide, using methyl red as the
indicator. Each cubic centimeter of fiftieth-normal acid is equivalent to
0.00578 Gm. of the alkaloids of Hyoscyamus."
The residual alkaloids, ~hich consist chiefly of the isomers

atropine and hyoscyamine, react with some of the 'sulfuric
acid as follows:
2C 17H 23 0 3N
289.3
+ H2S04~(C17H2303N2).H2S04
98
Since 98 Gm. of H 2S0 4 is equivalent to 2 X 289.19 Gm. of

atropine or hyoscyamine, each cubic centimeter of 0.1 N H 2S0 4
414
is equivalent to 2
~ ~ ~9i~000 =
0.02893 Gm. of the alkaloids
of belladonna. If exactly 10 Gm. of the drug is used as the

sample for assay, the percentage of alkaloids in the belladonna
cc. 0.1 N H 2S0 4 X 0.02893 X 100
may b e ca1cu1a t ed as f 0 11ows:
10
or cc. 0.1 N H 2S0 4 X 0.2893 = percentage of alkaloids in the
sample.
The U.S.P. requires that belladonna leaves contain not less
than 0.3 per cent of alkaloids. Calculate the percentage of
alkaloids contained in the sample assayed and compare the
results with the official requirements.
Belladonna leaf, belladonna root, and stramonium are assayed
by the same method as hyoscyamus with the exception that
smaller samplcs are used because these drugs contain larger
amounts of alkaloids.
Assay of Preparations of Hyoscyamus, Belladonna, and
Stramonium
Tinctures.-The tinctures of hyoscyamus, belladonna (leaf),
and stramonium of the U.S. Pharmacopoeia are all assayed by
the same method except that the amounts"'-of the tinctures
employed as samples are designated so that ap:proximately
0.030 Gm. of the alkaloids will be obtained in the assay residue.
The assay method is the same as that for hyoscyamus except
that the extraction of the alkaloids is omitted since they are
already in solution. The assay process is illustrated in the
following exercise.
Exercise 120
Object.-Assay of Tincture of Belladonna.

Materials Required.-100 cc. of tincture of belladonna.
250 cc. of ether.
Procedure.-Accurately measure 100 cc. of tincture of belladonna and
evaporate it, at a temperature not exceeding 80 a C., to a volume of about
10 cc. Add 10 cc. of approximately 0.1 N sulfuric acid and 10 cc. of distilled water. Filterthis mixture, collecting the filtrate in a separator, dissolve the chlorophyll residue in chloroform, add acidulated water, evaporate
on a water bath until the chloroform is removed, and filter into the same
separator through the filter previously used. Proceed as in Step 3 of the
Assay of Hyoscyamus (page 413).
415
Pilular and Powdered Extracts.-The pilular and powdered

extracts of hyoscyamus, belladonna, and stramonium are assayed
by the same method ~ith the exception that different amounts
of the extracts are used. The following procedures illustrate
the methods: assay of pilular extract of hyoscyamus.
"Dissolve approximately 5 Gm. of Pilular Extract of Hyoscyamus, accurately weighed (see Proximate Assays, page 395, for method of weighing),
in 10 cc. of chloroform, add 10 cc. of approximately tenth-normal sulfuric
acid and 40 cc. of distilled water, and evaporate the mixture on a water bath
until the chloroform is removed. Complete the assay as directed under
Hyoscyamus, page 412, beginning with the words 'Filter this mixture,
collecting the filtrate in a separator.' Each cubic 'centimeter of fiftiethnormal acid is equivalent to 0.00578 Gm. of the alkaloids of hyoscyamus."
Assay of powdered extract of
hyoscya~us.
"Digest approximately 5. Gm. of Powdered Extract of Hyoscyamus,

accurately weighed, in 10 cc. of chloroform, add 10 cc. of approximately
tenth-normal sulfuric acid and 40 cc. of distilled water. Mix well, filter
through purified cotton wetted with weak sulfuric acid, wash the cotton and
the residue with acidulated distilled water until free from alkaloids, discard
the cotton and the residue, and evaporate the filtrate on a water bath
until the chloroform is removed. Complete the assay as directed under
Hyoscyamus, page 412, beginning with the words 'Filter this mixture,
collecting the filtrate in a separator.' Each cubic centimeter of fiftiethnormal sulfuric acid is equivalent to 0.00578 Gm. of the alkaloids of
hyoscyamus. "
Fluidextracts.-Fluidextract of belladonna root of the U.S.

Pharmacopoeia is assayed by a method similar to that for
hyoscyamus as indicated by the following procedure:
"To 10 cc. of Fluidextract of Belladonna Root add 10 cc. of approximately
tenth-normal sulfuric acid and 10 cc. of distilled water, and evaporate on a
water bath to about 10 cc. Then add 10 cc. of distilled water and proceed
as directed under the Assay of Hyoscyamus, page 412, beginning with the
words 'Filter this mixture, collecting the filtrate in a separator.' Each cubic
centimeter of fiftieth-normal acid is equivalent to 0.00578 Gm. of the alkaloids of belladonna."
Fluidextracts of belladonna leaf, hyoscyamus, and stramonium

of the National Formular.y are all assayed by the same method.
The method described in the National Formulary differs somewhat from the method of the Pharmacopoeia as is shown in the
following exercise.
416

Exercise 121
Object.-Assay of Fluidextract of Belladonna Leaf.

Materials Required.-l0 cc. of fluidextract of belladonna leaf.
Procedure.-"To 20 cc. of chloroform in a separatory funnel, add 10 cc.
of Fluidextract of Belladonna Leaf, accurately measured, 10 ce. of water, and
about 1 cc. of ammonia water, and shake the mixture vigorously for one
minute. Separate the chloroformic layer, and complete the extraction of
the alkaloids with successive portions of chloroform. Extract the alkaloids
completely from the combined chloroformic solutions by shaking with successive small portions of dilute sulfuric Mid (about 1 per cent). Filter the
acid solutions successively through a small filter or pledget of cotton into a
separatory funnel. Add a slight excess of ammonia water, and completely
extract the alkaloids from the aqueous layer by shaking with successive'
TABLE LXII.-ALKALOIDAL ASSAYS BY THE TOTAL EXTRACTION PROCESS
Substance
U.S.P.
Belladonna leaf ................... .
Belladonna ointment .............. .
Belladonna, pilular extract of ...... .
Belladonna plaster ................ .
Belladonna, powdered extract of ... .
Belladonna root ................... .
Belladonna root, fluidextract of .... .
Belladonna, tincture of ............ .
Hyoscyamus ...................... .
Hyoscyamus, pilular extract of ..... .
Hyoscyamus, powdered extract of .. .
Hyoscyamus, tincture of ........... .
Stramonium ....... '............... .
Stramonium, pilular extract of ..... .
Stramonium, powdered extract of ...'
Stramonium, tincture of ........... .
N.F.
Belladonna leaf, fluidextract of ..... .
Hyoscyamus, fluidextract of . ...... .
Stramonium, fluidextract of ........ .
(n.l.t.)
not less than.
Amount
used,
Gm.or
cc.
Official
requirement,
per cent
10
25
3
10
2
10
10
100
25
5
5
250
10
3
2
100
(n.l.t.) l3
O. 118 0 O. 132
1.18 to 1.32
0.25 to 0.30
1.18 to 1.32
(n.l. t.) 0.45
0.405 to 0.495WjV
0.027 to 0.033WjV
(n.l.t.) 0.040
0.135 to 0.175
0.135 to 0.175
0.0034 to 0.0046WjV
(n.l.t.) 0.30
1.10 to 1.30
1.10 to 1.30
0.027 to 0.033WjV
10
25
10
0.27 toO.33W/V
0.035 to 0.045WjV
0.25 to 0.35WjV
417
portions of chloroform. Evaporate the combined chloroformic solutions to

dryness and heat the dry residue on a water bath for 15 minutes. Then &dd
about 1 cc. of neutral alcohol, again evaporate to dryness, and heat the residue on a water bath for 15 minutes. Mix the residue with 2 cc. of neutral
alcohol, add 15 cc. of fiftieth-normal sulfuric acid, and warm the mixture to
dissolve the alkaloids. Cool, and titrate the excess acid with fiftieth-normal
sodium hydroxide, using methyl red T.S. or cochineal T.S. as the indicator.
"Each cubic centimeter of fiftieth-normal sulfuric acid is equivalent to
0.005784 Gm. of the alkaloids of Belladonna Leaf."
Ointment of Belladonna and Belladonna Plaster.-Ointment

of belladonna and belladonna plaster of the U.S. Pharmacopoeia
are assayed by special methods designed to separate the alkaloids
from the ointment or plaster base. T):le principles involved are
similar to those aJ.ready considered.
1. What isomeric change does the chief alkaloid of belladonna undergo
in the assay process?
2. The alkaloids of belladonna have the structure of esters. What type
of decomposition might they undergo if allowed to stand in contact for a
long time with a strong alkali?
3. How much 0.1015 N H 2S0 4 will be required to form the sulfates of
the alkaloids from 25 Gm. of hyoscyamus leaves which contain 0.070 per
cent alkaloids?
CHAPTER XXVI
ALKALOIDAL ASSAYS BY SPECIAL METHODS
A number of the official alkaloidal drugs and their preparations

cannot be assayed by any of the type methods previously discussed because of the nature of the drug or of the properties of
the a1kaloids to be determined. Drugs of this type are assayed
by special methods.
The assays of opium and its preparations differ from those of
the other alkaloidal drugs, due to the fact that morphine exhibits
exceptional solubility properties; i.e., it is practically insoluble
in most organic solvents.
Exercise 122
Object.-Assay of Opium.
Materials Required.-6 Gm. of opium.
3 Gm. of freshly slaked lime.
1 G~. of ammonium chloride.
100 ce. of ether.
25 cc. of water saturated with morphine.
Procedure.-l. "Triturate in a mortar 6 Gm. of the Opium to be assayed
which, if fresh, should be in very small pieces, and if dry, in fine powder, with
about 40 cc. of distilled water for fifteen minutes. Transfer the mixture
completely to a flask with the aid of 30 cc. of warm distilled water, stopper
the flask, and shake it every ten minutes, or continuously in a mechanical
agitator, during one hour. Pour the contents as evenly as possible upon a
wetted filter of from 10 to 11 cm. in diameter or, preferably, upon a sintered
glass funnel attached to a suction flask. When the liquid has drained off,
wash the residue with abollt 20 cc. of distilled water, carefully dropped upon
the edges of the filter and its contents. Transfer the moist residue to a
mortar, rub it to a smooth paste, then rinse it into the original flask with
40 cc. of distilled water, agitate it thoroughly during ten minutes, and return
the entire mixture to the filter. When the liquid has drained off, wash the
residue with small portions of distilled water until the washings are nearly
colorless. "
The alkaloids of opium, about 25 of which are known, occur in

the drug as meconates and sulfates. When the opium is agitated
418
419
and macerated with distilled water, most of the meconates and

sulfates of the alkaloids are dissolved. Complete extraction
of the alkaloids is obtained when the residual opium is treated
with a second portion of distilled water. By rubbing to a smooth
paste, any lumps are broken up so that the water can come in
contact with all ~f the particles of the drug. The residue is
washed with distilled water after each extraction to displace all
of the solvent absorbed by the opium.
2. "Evaporate the filtrates and washings, in a tared dish, to about SO Gm.
and allow to cool. Now add 3 Gm. of freshly slaked lime, triturate the
mixture for fifteen minutes, and transfer it completely to a tared flask with
the aid of small portions of dii:ltilled water. Then add sufficient distilled
water to make the mixture weigh 54 Gm. and mix well. Filter the mixture
using a filter from 10 to 11 em. in diameter, which has not been previously
moistened, collecting the filtrate in a dry cylinder or small flask. During
the filtration keep the funnel-covered with a glass plate and its stem well
within the neck of the receiving vessel."
The combined aqueous filtrates are evaporated on a water

bath to a volume of about 30 cc., since the alkaloids which are
quite stable at 100C. might be decomposed if the evaporation
was carried out over a direct flame. The purpose of concentrating the aqueous alkaloidal extractive is to reduce the volume to
a convenient working quantity. The concentrated extract is
made up to a weight of exactly 54 Gm., so that an aliquot portion
representing a definite weight of opium may be taken. When
'the aqueous opium extract is triturated with freshly slaked
lime (prepared by adding dropwise 1 cc. of water to 3 Gm. of
quicklime, allowing the product to become pulverulent, and cool),
the alkaloids are liberated from their salts and the morphine,
codeine, and narcotine are dissolved in the alkaline calcium
hydroxide solution, most of the other alkaloids of opium remaining as insoluble matter along with the calcium meconate, calcium
sulfate, resins, etc. The mixture may be transferred to a flask
and shaken for 15 min. to allow the reactions in which the alkaloids are liberated to go to completion, the flask being stoppered
to prevent evaporation so that the volume of liquid will not
change appreciably, since an aliquot portion of it is employed in
the assay.
S. "Place 34 Gm. of this filtrate (corresponding to 4 Gm. of Opium) in an
Erlenmeyer flask of suitable capacity, add 2 ec. of alcohol and 15 cc. of ether,
420
QUANTITATI.VE PHARMACEUTICAL CHEMISTRY
and, after shaking the mixture, add 1 Gm. of ammonium chloride. Stopper
the flask, shake it frequently during ten minutes and set it aside in a cool
place over night. Remove the stopper and brush any adhering crystals back
into the flask."
Since the total aqueous extract containing the alkaloids

weighed 54 Gm. after adding the lime, 54 - 3 = the weight of
liquid and an aliquot portion of two-thirds of this liquid or 34 Gm.
contains ap. amount of the alkaloids corresponding to % of 6
4 Gm. of opium.
Ammonia is liberated from ammonium chloride:
and morphine is precipitated, since it is insoluble in ammoniacal

solution and in ether. The alcohol. causes the morphine to
separate out in the form of relatively large crystals; whereas,
when alcohol is not used the alkaloid comes down in the form of a
fine powder. The former are more easy to collect and less rapidly
soluble in ether than the powder. The mixture is allowed to
stand in a cool place for 12 hr. to allow the larger crystals to grow
at the expense of the smaller ones. The cod~ine and narcotine
remain in solution in the ether.
4. "Decant the ethereal layer through a small filter p~l'V.' ~~nse the
flask and contents with 15 cc. of ether and pass these washings through
the filter, and again wash the filter with an additional small quantity of
ether. When all of the ether has passed through the filter, pour the aqueous
layer upon the filter without trying to remove the crystals from the flask.
Wash the crystals in the flask and the contents of the filter with distilled
water, previously saturated with morphine, until the washings are colorless.
Then add a few drops of distilled water to displace the morphinated water."
The crystals are washed with ether to remove any traces of

codeine or narcotine, since these alkaloids are insoluble in ammonia water but soluble in ether. Water saturated with morphine
is used to wash the crystals to prevent solution of the crystalline
morphine which is very slightly soluble in water. This aqueous
solution removes any ammonia present as well as any other
water-soluble impurity adhering to the surface of the crystals,
such as gums, coloring matter, etc. The morphinated water
must be displaced from the paper filter with distilled water
421
to avoid including in the final result some of the morphine

from the morphinated water.
5. "Add to the flask containing the crystals about 10 cc. of hot neutral
methanol, agitate to dissolve as much of the morphine as possible, and pour
the solution over the morphine on the filter, receiving the filtrate in a suitable flask. Repel!-t this treatment with hot, neutral methanol three or four
times, using from 5 to 7 cc. each time, until all of the morphine has been dissolved. Cool this solution and add exactly 25 cc. of tenth-normal sulfuric
acid. Dilute this solution with 50 ee. of distilled water and titrate the
excess of acid with tenth-normal sodium hydroxide, using methyl red T.S.
as the indicator. Each cubic centimeter of tenth-normal acid is equivalent
to 0.02852 Gm. of anhydrous morphine."
The morphine crystals dissolve in the 0.1 N sulfuric acid with

the formation of morphine sulfate:
2C17H190aN + H2S0C~(C17H190aN)2.H2S04
2 X 285.16
98
One thousand cubic centimeters of N sulfuric acid is equivalent therefore to 285.16 Gm. of anhydrous morphine, and each
cubic centimeter of 0.1 N sulfuric acid is equivalent to
10 2~\~~00 = 0.02852 Gm. anhydrous morphine.

The Pharmacopoeia requires that opium contain not less than
9.5 per cent of anhydrous morphine. Calculate the percentage
of anhydrous morphine found in the sample of opium assayed
and compare the results with the official requirements.
Granulated opium and powdered opium are assayed in exactly
the same manner as described above. The general procedure
used in the assay of tincture of opium is also the same, except
that 60 cc. of the tincture is evaporated to remove the alcohol,
diluted to exactly 54 Gm. with distilled water, and treated with
lime, etc., as directed in steps 2 to 5 of the above assay procedure.
Camphorated tincture of opium is assayed by a special method.
Extract of opium is also assayed by this method, but in the first
step of the procedure, a 3 Gm." sample of the extract is dissolved
in water and the solution is diluted to 54 Gm. after the addition
of the lime, etc.
1. What functions do the calcium oxide and ammonium chloride serve
in the above assay?
422
2. One gram of morphine is soluble in about 7,632 cc. of ether at 25C.

Look up the solubility of codeine in the U.S.P. and explain how ether may
be used to separate the alkaloids.
3. Why is water saturated with morphine used to wash the crystallized
morphine?
4. Morphine contains a phenolic hydroxyl group. Explain why it is
appreciably soluble in alkaline solutions.
TABLE LXIII.-OFFICIAL SUBSTANCES ASSAYED BY THE OPIUM ASSAY
METHOD
Amount
Equivalent
used,
of 1 cc. of
Gm. or
0.1 N H 2SO 4
cc.
Substance
Anhydrous morphine, official

requirement,
per cent
U.S.P.
Opium ............. , ....
Opium, granulated ...... "
Opium, powdered .........
Opium, tincture of ........
Opium, tincture of, camphorated ..............
6
6
6
60
0.02852
0.02852
0.02852
0.02852
100
0.00570*
9.5
10
to 10.5
10
to 10.5
0.95 to 1.05W/V
0.035 to 0.0045
N.F.
Opium, extract ...... , ....
* Equivalent of 1
CC.
0.02852
19.~
.....
to 20.5
of 0.02 N H 2SO
Exercise 123
Object.-Assay of Colchicum Seed.

Materials Required.-15 Gm. of colchicum seed in No. 60 powder.
10 cc. of solution of lead subacetate.
2 Gm. of sodium phosphate.
20 cc. of ether.
Procedure.-l. "Place 15 Gm. of Colchicum Seed, in fine powder, in ?oJ
suitable flask and add 290 cc: of distilled water and 10 cc. of lead subacetate
T.S. Weigh the flask and contents, and digest the mixture at from 60 to
70C. for three hours with occasional agitation. Cool, add distilled wat!)r
to restore the original weight, and filter, collecting 200 cc. of filtrate."
Colchicine is the only medicinally important constituent of

colchicum. Both the seed and corm, which are assayed by
'the same method, contain a considerable amount of starch,
sugar, gum, tannin, resin, fatty oil, and coloring matter from
423
which the alkaloid must be separated. The free alkaloid, colchicine, is miscible in all proportions with water, and it is extracted
from the drug when the latter is digested with water. The
digestion temperature should not exceed 70C., because the
starch content of the drug, especially of the corm, is gelatinized
at higher temperatlJres, and because colchicine is less soluble in
hot than in cold water. The lead subacetate is added to prevent
solution of the tannin, gums, resin, and coloring matter with
which it forms water-insoluble compounds and to liberate the
colchicine from acids with which it may be combined in the crude
drug. The mixture is cooled, and distilled water is added to
restore the original weight of the sample, water, and lead subacetate mixture, so that an aliquot portion, corresponding to a
definite weight of the sample taken, may be used. Two hundred
cubic centimeters of the liquid representing the alkaloids contained in two-thirds of the weight of the sample are filtered off.
2. "Add 2 Gm. of sodium phosphate to the clear filtrate, or sufficient to
precipitate the lead completely, shake the mixture frequently during half an
hour, ffiter, collect 100 cc. of ffitrate, representing 5 Gm. of Colchicum Seed."
The sodium phosphate precipitates the excess lead subacetate

from the solution as insoluble lead pl).osphate, leaving sodium
acetate in solution. A second aliquot portion of 100 cc. corresponding to one-third of the total sample used or 5 Gm. of colchicum is taken.
3. "Completely extract the alkaloid from the ffitrate with chloroform, as
shown by testing with iodine T.S., and evaporate the solution in a tared
flask. Add 1 cc. of alcohol, and again evaporate. Repeat this operation
once more, and dry the residue to constant weight at 100C."
Colchicine is miscible in all proportions with chloroform, and

this solvent extracts the alkaloid, which is very feebly basic,
from neutral or acid solutions. The alkaloid forms an addition
product of unknown constitutio~ of the formula C22H2606N.2CHCla with chloroform. The chloroform is driven off from this
addition product when it is treated with successive portions of
alcohol and evaporated to dryness at 100C. The residue, dried
to constant weight at 100C. and weighed accurately, consists
chiefly of crude colchicine and colchiceine.
424
4. "To this weighed residue in the flask, add 5 cc. of tenth-normal sulfuric
acid, 5 cc. of distilled water, and a few drops of chloroform, and heat the
mixture at 70C. for ten minutes. Filter the liquid through a pledget of
purified cotton, wash the flask and cotton with distilled water, reject the
filtrate and washings, and remove as much of the water from the cotton as
possible. Dissolve the residue, if any, which may remain on the cotton by
washing it first with a little alcohol and then with ether; collect the alcoholether washings in the flask, evaporate, and dry the residue to constant
weight at 100C. Deduct this weight from the weight of residue previously
obtained. The difference is the weight of colchicine obtained from 5 Gm.
of CoI<:hicum Seed."
A dilute solution of sulfuric acid heated to 70C. is used to

dissolve the colchicine completely from the residue, since this
alkaloid decomposes rapidly when heated to 100C. with strong
mineral acids. The addition of a few drops of chloroform
facilitates the solution of the colchicine from the residue. The
colchicine in acid solution is filtered off through cotton and
removed from the filtration medium by washing with distilled
water, leaving acid-insoluble impurities on the cotton and in the
flask. The impurities are dissolved by successive treatment
with alcohol which is miscible with water and with ether. The
total residual impurities, when dried to constant weight at the
same temperature as the crude colchicine and <ieducted from
the weight of the latter previously obtained, gives the weight of
colchicine which dissolved in the acid solution; e.g., if an aliquot
portion corresponding to 5 Gm. of colchicum yields 0;'025 Gm. of
crude colchicine and the residual impurity is found to weigh
0.025 - 0.005
00
h .
O.005 Gm., t h en
5
X 1 = 0.4 per cent colc !ClUe
contained in the sample.
Colchicum seed must contain 0.45 per cent of colchicine to
meet the official requirement. Calculate the percentage of
colchicine contained in the sample assayed and compare the
result with the official requirement.
Colchicum corm is assayed in the same way as colchicum seed.
The extract, fluidextract, and tincture are also assayed by the
same method, but in the case of the fluidextract, 15 cc. is taken
as the sample and 275 cc. of water is used to bring the volume of
the liquid to 290 cc., so that when 10 cc. of lead sub acetate is
added, the total volume of the mixture will be 300 cc. In the
425
assay of the tincture, a volume of 150 ce. is taken, evaporated to

15 cc., and made up to 300 cc. by the addition of 275 cc. of distilled water and 10 cc. of lead subacetate solution.
Questions !lad Problems
1. What exceptional solubility properties are exhibited by colchicine?
2. Why is lead subacetate used in the extraction of the alkaloid from
the crude drug?
3. Why should not the aqueous mixture be heated above 70C. in the
extraction of colchicine from the powdered drug?
4. What property of colchicine makes it impossible to determine this
alkaloid accurately by volumetric methods?
5. Why should the crude colchicine not be heated above 70C. with the
0.1 N sulfuric acid?
TABLE LXIV.-OFFICUL SUlISTANCES ASSAYED BY TIlE SAME METIIOD AS
COLCHICUM
Substance
Amount
used
U.S.P.
Colchicum seed ............ 15 Gm.
Colchicum seed, tincture of 150 cc.
Aliquot portion corresponds to
Colchicine, official
requirement,
per cent
5 Gm.
50 cc.
0.45
0.036 to 0.044W IV
N.F.
Colchicum corm ...........
Colchicum corm, fluidextract of. ................
Colchicum corm, strong
tincture of ..............
Colchicum seed, fluidextract
of .....................
15 Gm.
5Gm.
0.35
15 cc.
5 cc.
0.31 to 0.39WIV
45 cc.
15 cc.
0.125 to 0.155WIV
15 cc.
5 cc.
0.4
toO.5WIV
Exercise 124
Object.-Assay of Nux Vomica.

Materials Required.-10 Gm. of nux vomica in No. 60 powder.
100 cc. of ether-chloroform mixture.
Procedure.-l. "Place 15 Gm. of Nux Vomica, in coarse powder, in a
flask or bottle, add 150 cc. of a mixture of 3 volumes of ether and 1 volume
of chloroform, agitate the mixture, and allow it to stand for about two
minutes. Then add 10 cc. of stronger -ammonia T.S., agitate thoroughly,
stopper the container securely and shake frequently, but gently, during one
426
hour. Now allow the mixture to stand for twelve hours or over night in a
cool place, At the expiration of this period, shake the container gep.tly for
fifteen minutes, and then allow the liquids to separate."
The alkaloids of nux vomica which consist principally of

strychnine and brucine are set free by the ammonia water and
dissolve in the ether-chloroform mixture,
2. "Decant 100 cc. of the liquid (representing 10 Gm. of Nux Vomica),
and transfer it to a separator, rinsing the container with a little chloroform
and adding the rinsings to the separator. Now add about 40 cc. of approximately normal sulfuric acid to the separator and shake the mixture gently
for five minutes, then allow the liquids to separate, and draw off the acid
layer into another separator and repeat with successive portions of the acid,
until the ethereal liquid is completely extracted." (Test for the complete
extraction of the alkaloids as directed on page 399.)
The alkaloidal sulfates are formed and dissolve in the acid

solution; fats, resins, coloring matter, etc., remain in the etherchloroform layer.
3. "To the combined acid solutions in the separator, add a small piece of
red litmus paper and 50 cc. of chloroform, and follow with sufficient ammonia
T.S. to render the aqueous layer alkaline and, after gently shaking, add 2 or
3 cc. more of the ammonia T.S. Now shake the mixtUre thoroughly, but
gently, for about ten minutes, and allow the liquidS1to separate. Draw off
the chloroform into a container and repeat, with Ildditional\ portions of
chloroform, until all of the alkaloid is extracted."
-
The alkaloids are liberated and dissolve in the chlo:r;oform.

4. "Carefully evaporate the combined chloroformic extracts to dryness on
a water bath, dissolve the residue by warming with 15 cc. of approximately
3 per cent sulfuric acid, cool, and then add 3 cc. of a mixture of equal parts
of nitric acid and a 5 per cent solution of sodium nitrite in distilled water,
stir well, and allow to stand for exactly ten minutes at room temperature.
At the expiration of this period, pour the red solution into a separator containing 50 cc. of chloroform, rinse the flask with distilled water, and add the
rinsings to the separator-. Now immediately add sufficient 10 per cent
sodium hydroxide solution to the contents of the separator to render it
distinctly alkaline to litmus paper, and then add a few cubic centimeters
more of the hydroxide solution."
The strychnine and brucine are converted to their sulfates by

the sulfuric acid. When the nitric acid and sodium nitrite mixture is added, the brucine is oxidized to a non-alkaloidal, red
colored, quinonoid compound having phenolic properties. The
427
strychnine is very stable to oxidizing agents and is not attacked

in significant amounts during the short period of oxidation. The
decomposition of the brucine is completed in 10 min. If the
mixture were allowed to stand for a longer period, a small amount
of the strychnine would be oxidized. Consequently, the mixture
is neutralized with sodium hydroxide to stop the oxidation and
an excess of the alkali is added to liberate the strychnine. Some
workers prefer to liberate the strychnine by the addition of alkali
prior to the addition of the organic solvent because the complete
decomposition of the strychnine salts is difficult to accomplish
in the presence of organic solvents.
5. "Shake the mixture gently for ten minutes and allow the liquids to
separate. Draw off the chloroformic layer into another separator and
repeat the shaking out with additional portions of chloroform until the
alkaloid is completely extracted. Add 10 cc. of distilled water to the combined chloroformic extract, shake the mixture gently, and add a small piece
of red litmus paper. The litmus paper should indicate not more than a slight
alkalinity. If the water, after shaking with the chloroform, is strongly alkaline, draw off the chloroform into another separator, and shake it with another
10 cc. of distilled water. Draw off the chloroform, passing it through a filter
paper moistened with chloroform, into a container. Wash the filter paper
with warm chloroform and add it also to the container. Now shake out the
combined water extract with 5 cc. of chloroform and draw off all of the
chloroform through a chloroform-moistened filter paper as before."
The mixture is shaken for 10 min. to completely decompose the

strychnine salts. The liberated strychnine dissolves in the
chloroform while the phenolic quinonoid oxidation product from
the brucine remains in the aqueous alkaline layer. Sodium
hydroxide rather than ammonium hydroxide is used as the alkali
because ammonium hydroxide is not It'sufficiently strong base to
combine completely with the phenolic oxidation products and
prevent their passing into the chloroform. The combined
chloroform extractive is washed with water to remove any alkali
or alkali derivative of the phenolic oxidation products. This
aqueous wash liquid is then washed with chloroform to recover
any trace of strychnine.
6. "Evaporate the combined chloroform very carefully on a water bath
nearly but not quite, to dryness. Add to the moist residue 6 cc. of tenthnormal sulfuric acid and follow by 30 co. of distilled water. Heat the mixture
on a water bath until the alkaloid is dissolved and the odor of chloroform is
428
.QUANTITATIVE PHARMACEU,/,ICAL CHEMISTRY
dissipated. Cool to room temperature and titrate the excess of acid with
fiftieth-normal sodium hydroxide, using 1 drop of methyl red T:S. as the
indicator. Each cubic centimeter of tenth-normal sulfuric acid is equivalent
to 0.03342 Gm. of strychnine."
The chloroform should not be evaporated to dryness before

adding the acid since there is likely.to be some loss of strychnine
by decrepitation if this is done. The strychnine is converted into
its sulfate quantitatively by the 0.1 N H 2S0 4 ; the excess acid is
then determined by titration with 0.020 N NaOH. The equation
for the reaction is:
+ H2S04---+(C21H2202N2)2.H2S04
2C21H2202N2
334.19
The number of cubic centimeters of 0.1 N H 2S0 4 found upon

residual titration with 0.020 N NaOH subtracted from the
number of cubic centimeters of standard acid solution originally
added gives the quantity of standard acid solution which combined with the alkaloids.
The U.S.P. requires that nux --vomica contain not less than
2.5 per cent of alkaloids. Calculat~he pe~centage of alkaloids
contained in the sample assayed and compare the result with the
I
TABLE LXV.-ASSAYS OF
Nux
VOMICA AND
Amount
used,
Gm. or
cc.
Substance
U.S.P.
Nux
Nux
Nux
N.F.
Nux
Nux
vomica ................ '.. .

vomica, extract 9f. ....... .
vomica, tincture of ....... .
vomica, fluidextract of .....
VOMICA PREPARATIONS
Official'requirement,
per cent
15
Strychnine = (n.Lt.) 1.15
1.5 Strychnine = 7 to 7.75
100
Strychnine = 0.108 to 0.120
10
Strychnine = 1.05 to 1.25
(n.l.t.) = not less than.

1. Why must care be used to prevent emulsification when shaking out

the ether-chloroform mixture with acid solution?
429
2. Explain the chemical changes which the strychnine undergoes during

the various steps of the assay process.
3. Why is the deterlhination of the total alkaloids of nux vomica not a
correct measure of the therapeutic value of the drug?
4. If the total alkaloids of nux vomica were known to consist of strychnine
and brucine in the ratio 3: 1, what would be the proper equivalent to use in
their volumetric estimation?
Assay of Caffeine-containing Drugs.-Caffeine is the chief

alkaloidal constituent of a number of drugs used in different parts
of the world for beverage purposes such as tea, mate, coffee,
kola, and guarana, the last two of which are official. In the
determination of the caffeine content of the official drugs, it
should be borne in mind that caffeine differs in its properties
from most other alkaloids in the following ways: (1) It is very
feebly basic and cannot be determined by the volumetric method.
(2) The free alkaloid is appreciably soluble in warm water
(1 Gm. is soluble in 5.5 cc. at 80C.). (3) It can be shaken out of
acid solution readily with chloroform. (4) It is not precipitated
from solution by mercuric potassium iodide. (5) It sublimes
without decomposition at about 180C.
The assays of guarana and kola are performed by the same
method.
Exercise 125
Object.-Assay of Guarana.
Materials Required.~ Gm. of guarana in No. 60 powder.
Procedure.-l. "Place 6 Gm. of Guarana, in fine powder and accurately
weighed, into a suitable container, add 120 cc. of chloroform, allow the
mixture to stand about five minutes, and then add 6 cc. of ammonia water
and 6 cc. of distilled water. Shake t.he mixture continuously for one hour
or intermittently dUring two hours, and allow it to stand overnight.
Again shake intermittently during one half hour and separate 100 cc. of
the clear chloroformic solution."
That portion of the caffeine which occurs as a salt of tannic

acid is liberated by the ammonia and is dissolved in the chloroform along with the caffeine which occurs in the free state.
2. "Evaporate the chloroform, and treat the residue with 10 cc. of approximately 1 per cent sulfuric acid with the aid of a gentle heat. Filter the
cooled solution into a separator, and wash the dish and filter with successive
430
small portions of distilled water until no test for alkaloid is obtained with
iodine T.S. in a portion (1 cc.) of the filtrate after a:cidulating strongly with
diluted sulfuric acid."
The aliquot portion of the chloroformic solution is evaporated

to dryness so that the alkaloid may be dissolved in acid. This
end is very difficult to accomplish by shaking out the chloroform
solution with acid since chloroform removes caffeine from acid
solution. The residue should not be dried at a temperature exceeding 100C. because caffeine sublimes at higher
temperatures.
When the resi,due consisting of fat, coloring matter, etc., and
free alkaloid is treated with dilute sulfuric acid, the alkaloidal
sulfate dissolves in the aqueous solution, leaving the waterinsoluble impurities. Caffeine is precipitated from an acidulated,
but not from a neutral or alkaline aqueous solution by iodine
test solution.
3. "Make the combined filtrates alkaline with ammonia water and again
extract t.he alkaloid completely with successive portions of chloroform.
Evaporate the combined chloroformic solutions nearly to dryness, add about
1 cc. of neutral alcohol, and continue evaporation j;o dryness. Dry the
residue to constant weight at 100C.
"The weight obtained represents the yield of anhydrous laffeiqe, C 8H 1002N., from 5 Gm. of Guarana."
The alkaloid liberated from its sulfate by the ammonia is

dissolved in the chloroform. Caffeine is determined gravimetrically, since it is a very feeble base which does not give a distinct
end point with any indicator upon titration.
The fiuidextracts of kola and guarana are made alkaline with
ammonia and extracted directly with chloroform. The dried
residue from the chloroformic solution is then treated with warm
dilute acid to dissolve the free alkaloid and the alkaloid is
shaken out from the aqueous solution with chloroform and
determined gravimetrically in the same way as in the assay of
the crude drug.
1. Why is not the caffeine extracted from the chloroformic solution with
dilute acid in the usual manner?
2. Could mercuric potassium iodide solution be used to test for the complete extraction ,of the caffeine from the acid solution?
431
3. Why should caffeine be dried at a low temperature?

4. Explain why warm water can be used to dissolve the free alkaloid from
the chloroformic residue in the assay of the fluidextract of kola.
TABLE LXVI.-OFFICIAL DRUGS AND GALENICAL PREPARATIONS ASSAYED
FOR CAFFEINE
Amount
used,
Gm. or
per cent caffeine
cc.
Substance
N.F.
Guarana .................
Guarana, fluidextract of ...
Kola ...................
Kola, fluidextract of .......
.
.
.
.
6
5
12
10
4.0
3.6 to 4.4 W IV
1.0
0.85 to 1.15W IV
Assay of Alkaloidal Salts.-The determination of the amount

of alkaloid present in an alkaloidal salt or in a simple preparation
containing an alkaloid or an alkaloidal salt is generally a relatively
simple process since the alkaloid does not need to be freed from
a number of organic impurities as is the case in the assay of crude
drugs. The solvents used and the gravimetric or volumetric
methods employed differ according to the characteristic properties of the alkaloid determined.
Exercise 126
Object.-Assay of Citrated Caffeine.

Materials Required.-l Gm. of citrated caffeine.
Procedure.-" Accurately weigh about 1 Gm. of Citrated Caffeine, previously dried to constant weight at 80C., and dissolve it In 10 cc. of hot
distilled water; add 8 cc. of sodium hydroxide T.S., cool the solution, and
shake it in a separator with three or more successive portions of 20 cc. each
of chloroform, to effect complete -extraction. Filter the combined chloroformic solutions through a small filter, previously moistened with chloroform,
into a tared porcelain dish. Wash the filter and funnel with 10 cc. of hot
chloroform, adding the rinsings to the dish, and evaporate the combined
chloroformic solutions on a water bath, adding 2 cc. of alcohol just before the
last trace of chloroform is expelled. Complete the evaporation of the
solvent and dry the residue, consisting of anhydrous caffeine, to constant
weight at 80C."
432
The citrated caffeine is dried before it is weighed to drive off

moisture. Since caffeine is a very weak base and ~itric acid is
feebly acidic, it is probable that the aqueous solution of citrated
caffeine is dissociated almost completely into caffeine and citric
acid. Citric acid is quite soluble in chloroform, but its sodium
salt is insoluble in this solvent. The chloroform, therefore, dissolves the caffeine, leaving the sodium citrate in the aqueous
solution.
Exercise 127
Object.-Assay of Eucaine Hydrochloride.

Materials Required.-0.5 Gm. of eucaine hydrochloride.
100 cc. of neutral alcohol.
Procedure.-Dissolve about 0.5 Gm. of eucaine hydrochloride, previously dried to constant weig -'itt 100C. and accurately weighed, in
100 cc. of neutral alcohol. Titrate this alcoholic solution with 0.1 N sodium
hydroxide, using phenolphthalein T.S. as indicator. Each cubic centimeter
of 0.1 N sodium hydroxide corresponds to 0.02836 Gm. of C 5H 7 N(CH,),O(CsH.CO).HCl.
Neutral alcohol is specified because the ordinary commercial

alcohol frequently contains considerable amounts of acetic acid
which, if present, would react with the standard alkali. The
standard alkali solution reacts quantitatively with the alkaloidal
salt liberating eucaine:
C6H7N(CHa)aO(C6H5CO).HCl
283.64
+ NaOH----+NaCI + H
C5H7N(CH3)30(C6H5CO)
The liberated alkaloid is dissolved in the alcohol.
Each cubic
centimeter of 0.1 N NaOH corresponds to 10 2~\~00
0.02836
Gm. eucaine hydrochloride.

Exercise 128
Object.-Assay of Theobromine with Sodium Salicylate for

Theobromine.
Materials Required.-2 Gm. of theobromine with sodium salicylate dried
to constant weight at noC.

Procedure.-l. "Dissolve about 2 Gm. of Theobromine with Sodium
Salicylate, previously dried to constant weight at nOo. and accurately
433
weighed, in 10 cc. of warm distilled water, and titrate the solution with
normal hydrochloric acid, using phenolphthalein T.S. as the indicator: not
more than 5.5 cc. of normal hydrochloric acid is required to neutralize 2 Gm.
of the dried Theobromine with Sodium Salicylate."
Sodium theobromine is an unstable compound which is formed

when the alkaloid is dissolved in a solution of sodium hydroxide.
A solution of sodium theobromine is strongly 11lkaline, and
the alkali may be titrated directly with standard acid, the theobromine being precipitated at the same time. In the process
of manufacture of theobromine with sodium salicylate, a slight
excess of alkali is added. If only the theoretical quantity of
alkali were used, sufficient carbon dioxide would be absorbed
during the powdering and drying to make the product incompletely soluble in water. A limit of titratable alkali is necessary
to prevent the use of products containing too large an excess.
In the titration, only the sodium combined with the theobromine
and that in excess is determined since the titration is stopped
at the phenolphthalein end point which is on the alkaline side
of the pH scale. The sodium salicylate does not enter into the
reaction.
2. "The solution should now be slightly alkaline to litJIluS paper or be
made so by the addition of 1 or 2 drops of very dilute ammonia T.S. Allow
to stand at from 20 to 25C. for three hours, stirring occasionally, transfer
the precipitate of theobromine to a dried and weighed filter of 9 cm. diameter,
wash the precipitate and filter with four successive portions of 5 cc. each
of ice-cold distilled water, and dry to constant weight at 100C. To the
weight of precipitate thus obtainelf add 0.13 Gm. (the approximate quantity
of theobromine remaining in the liquid and washings). The total weight
corresponds to not less than 46.5 per cent of the weight of dried Theobromine
with Sodium Salicylate taken for the assay."
The theobromine is precipitated from the alkaline solution,

collected, washed, dried, and weighed. Since theobromine is
slightly soluble in the liqtQd from which it is precipitated, a correction is applied to the result.
It is difficult, to separate the theobromine from the mixture
because acidification liberates both the theobromine and the
salicylic acid and both of these compounds form soluble products
with excess of alkali. Theobromine is practically insoluble in all
of the common organic solvents and therefore cannot be extracted
434
in the usual manner. A better method of assay of the ~heo

bromine with sodium salicylate than the official method is one
which is based upon the conversion of the theobromine, 3: 7
dimethyl-xanthine, into caffeine, 1: 3: 7 trimethyl-xanthine, by
a procedure similar to that given for the assay of theophylline
with sodium acetate.
Exercise 129
Object.-Assay of Theophylline with Sodium Acetate for

Theophylline.
Materials Required.-l Gm. of theophylline with sodium acetate.
0.8 cc. of dimethyl sulfate.
Procedure.-l. "Disperse about 1 Gm. of Theophylline with Sodium
Acetate, accurately weighed, in 5 cc. of distilled water and 4.5 ce. of normal
sodium hydroxide, in a small stoppered flask. Add 0.8 cc. of dimethyl
sulfate, shake the mixture until solution is complete, and set it aside for one
hour, shaking it occasionally. Then add 5 cc. of normal sodium hydroxide
and 10 cc. of distilled water and shake the mixture for one minute."
The theophylline, 1: 3 dimethyl-xanthine is converted quantitatively by methylation into caffeine, 1: 3: 7 trimethyltxanthine,

when the dissolved sample is shaken with dimetQyl sulfate in
the presence of sodium hydroxide:
2. "Transfer it to a separator with the aid of chloroform and a little

distilled water, and extract the caffeine thus formed immediately by shaking
the liquid successively with 10-, 10-, 10- and 5-cc. portions of chloroform,
passing each ehloroformic solution through a small filter, previously moistened with chloroform, into a tared porcelain dish, finally washing the stem
of the separator and the filter with 10 cc. of hot chloroform. Evaporate the
combined chloroformic solutions on a water bath, adding 2 cc. of alcohol
before the last traces of chloroform are expelled. Complete the evaporation
of the solvent, and dry the residue, consisting of anhydrous caffeine, to
constant weight at 80C.: the weight of anhydrous caffeine so obtained,
multiplied by 0.9287, indicates the corresponding weight of anhydrous
theophylline. "
The caffeine formed, unlike theophylline, does not form a

soluble product with alkali and can be extracted with chloroform.
ALKALOIDAL ASSAYS BY SPEOIAL METHODS
435
Exercise 130
Object.-Assay of Ephedrine Sulfate.

Materials Required.-O.5 Gm. of ephedrine sulfate.
100 cc. of ether.
10 cc. of alcohol (neutralized to bromthymol blue).
Procedure.-l. "Transfer about 0.5 Gm. of Ephedrine Sulfate, dried over
sulfuric acid for twenty-four hours and accurately weighed, to a separator;
add 10 cc. of distilled water and 5 cc. of ammonia T.S., and extract with six
portions of ether, using 25 ce., 20 ee., 15 ec., 15 ee., 15 cc., and 15 cc.,
respectively."
The ephedrine liberated from the sulfate by the ammonia is

extracted by the ether. Chloroform cannot be used to shake out
the ephedrine because this alkaloid is a sufficiently strong base to
decompose the chloroform and form ephedrine hydrochloride
especially in warm solution. (See test for identity of ephedrine,
U.S.P., page 144.)
2. "Collect the extracts in a beaker, evaporate the ether on a water bath
to about 5 cc., add 10 cc. of alcohol, previously neutralized to bromthymol
blue T.S. with tenth-normal sodium hydroxide, and add from a burette an
excess of tenth-normal hydrochloric acid. Titrate the excess of acid with
tenth-normal sodium hydroxide, using bromthymol blue T.S. as the indicator. The ephedrine thus found is not less than 75.5 and not more than
77.3 per cent of the weight of Ephedrine Sulfate taken. One cubic centimeter of tenth-normal hydrochloric acid is equivalent to 0.01651 Gm. of
ephedrine.' ,
The extracted ephedrine is dissolved by the alcohol and converted quantitatively to ephedrine hydrochloride by the acid:
C 1oH 150N
165.13
+ HCl~ClOH150N.HCl
201.60
436
TABLE LXVII.-ASSAYS OF ALKALOIDS, ALKALOIDAL SALTS,. AND SXMPLE

PREPARATIONS
Substance
U.S.P.
Caffeine, citra ted ............... .
Caffeine with Bodium benzoate .. . .
Codeine phosphate ............. .
Ephedrine ..................... .
Ephedrine hydrochloride ........ .
Ephedrine Bulfate . ... " ......... .
Ethylhydrocupreine hydrochloride
Eucaine hydrochloride .......... .
Phenacaine hydrochloride ....... .
Quinine and urea hydrochloride . . .
Theobromine with sodium salicylate ......................... .
Theophylline with ethylene diamine ........................ .
Amount
used,
Gm. or
cc.
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
Anhydrous caffeine = 48 to 52
Anhydrous caffeine = 47 to 50
Anhydrous codeine = (n.l.t.) 70
Ephedrine = 98 to 100
Anhydrous ephedrine = 80 to 82.5
Anhydrous ephedrine - '(:5.5 to 77.3
Ethylhydrocupreine = (n.l. t.) 90
Eucaine hydrochloride = (n.l.t.) 99
Phenacaine = 87.5 to 90.5
Anhydrous quinine = 58 to 65
Theobromine = (n.l.t.) 46.5
Anhydrous, theophylline = 70 to 80
Anhydrous theophylline = 55 to 65
1
1
Theophylline with sodium acetate.

1
N.F.
Apomorphine hydrochloride, tab0.00
lete of ......................
Atropine sulfate, tahlets of
containing 0.02 or more Gm ..
0.08
containing 0.0012 to 0.02 Gm ..
0.08
containing less than 0.0012 Gm. 0.08
Caffeine citrated, tablete of ...... . 0.5
Caffeine with sodium benzoate,
ampule of. .................. . 0.5
Caffeine with sodium benzoate,
tablete of. ................... . 0.5
Cocaine hydrochloride, tablets of ..
0.06
Codeine phosphate, tablets of. ....
0.06
Codeine sulfate, tablets of. ......
0.06
Emetine hydrochloride, ampuls of
0.3
Ephedrine hydrochloride, tablets of
0.06
Ephedrine sulfate, ampul. of ..... . 0.5
Ephedrine sulfate, solution of. ... . 10
Morphine sulfate. tablets of ...... . 0.13
Morphine and atropine sulfates,
tablets of. ...................
0.13
Nux vomica alkaloids, solution of . . 20
Procaine hydrochloride, ampuls of
Procaine hydrochloride, solution of
Procaine hydrochloride, tablets of.
Quinine dihydrochloride, ampuls of
Quinine hydrochloride and ethyl
carbamate, ampule of . ........ .
Quinine and urea hydrochlorides,
ampuls of. .................
Quinine phosphate. . ........... .
Quinine sulfate, tablets of ...... .
Scopolamine hydrobromide, tablets
of. ......................
containing 0.0012 Gm. or more.
containing less than 0.0012 Gm
Strychnine nitrate, tablets of .... .
containing O. 02 Gm. or more .. .
containin,,; 0.0012 to 0.02 Gm.
containing less than O. 0012 Gm.
Strychnine phosphate ...........
Strychnine sulfate, tablets of .... .
containing 0.02 Gm. or more .. .
containing O. 0012 to O. 02 Gm ..
containing less than O. 0012 Gm.
(n.l.t.) = not less than .
Amount of equivalent sought.
0.2
10
0.2
0.5
Apomorphine hydrochloride = 91 to 109'

Atropine
Atropine
Atropine
Caffeine,
sulfate .. 92.5 to 107. 5b

sulfate .. 91 to 109b
sulfate .. 88 to 112'
citra ted = 43 to 53 b
Anhydrous caffeive
= 45 to 52b
Anhydrous caffeine = 43.5 to 53. 5b

Cocaine hyprochloride = 91 to 109b
Anhydrous codeive = 63 to 77'
Codeine sulfate = 91 to 109/>
Emetine hydrochloride = 84 to 92 b
Ephedrine hydroohl6ride = 91 to 109'
Ephedrine = 72.6 to 80. 2b
Ephedrine sulfate = 2.8 to 3. 2W IV
Morphine sulfate = 92.5 to 107. 5b
Morphine sulfate = 91 to 109b
Strychnine sulfate = 1. 5 to 1.7 W IV
Brucine sulfate = 1. 5 to 1. 7 W IV
Procaine hydrochloride = 95 to 105b

Procaine hydrochloride = 1.9 to 2.1 W IV
Procaine hydrochloride = 92.5 to 107.5'
Quinine dihydrochloride = 95 to 105b
0.5
Quinine hydrochloride = 92.5 to 107. 5 b
0.5
Quinine and urea hydrochlorides = 54.8 to

63.7
Anhydrous quinine = 74 to 78
Quinine sulfate = 91 to 109b
0.5
0.13
0.06
0.13
0.4
0.13
Scopolamine hydrobromide = 91 to 109b

Scopolamine hydrobromide = 88 to 112 b
Strychnine
Strychnine
Strychnine
Strychnine
nitrate = 92.5 to 107. 5b

nitrate = 91 to 109b
nitrate = 88 to 112b
= 70 to 73
Strychnine sulfate = 92.5 to 107. 5b

Strychnine sulfate = 91 to 109b
Strychnine sulfate = 88 to 112 b
CHAPTER XXVII
OTHER OFFICIAL ASSAYS INVOLVING THE USE OF
IMMISCIBLE SOLVENTS
Use is made of the immiscible solvent principle of extraction
and purification in the proximate determination of the amount
of active constituent contained in a number of non-alkaloidal
drugs such as jalap, cantharides, and oleoresin of aspidium and
in the assay of certain synthetic medicinal products, e.g., soluble
barbital.
Exercise 131
Object.-Assay of Cantharides.
Materials Required.-15 Gm. of cantharides in No. 40 powder.
75 cc. of purified petroleum benzin.
100 cc. of benzene.
25 cc. of absolute alcohol.
Procedure.-l. "Place 15 Gm. of Cantharides, in moderately coarse
powder, in a pressure bottle' of not less than 250 cc. capacity, add 150 cc. of a
mixture of benzene, two volumes, and purified petroleum benzin one volume,
and then add 2 cc. of hydrochloric acid. Stopper the bottle tightly, shake
it well, and allow it to stand about ten hours. Now gradually warm the
bottle and its eontents to about 40C. and maintain it at approximately
that temperature with frequent shaking during three hours, avoiding
evaporation. If necessary, add more solvent to replace any lost by
evaporation."
Cantharidin, C sH 120:(CO)2:0, is the inner anhydride, lactone,

of the dibasic cantharidic acid, C SH 120(COOH)2. Both cantharidin and the alkali salts of cantharidic acid occur in the
beetles. When the above mixture is acidulated, the cantharidin
and the liberated cantharidic acid dissolve in the warm benzenepetroleum benzin mixture together with fats, coloring matter,
and other inactive constituents. Although a Soxhlet apparatus
can be used to advantage in this assay, the official directions are
to extract the drug by maceration, the purpose being to adapt
437
438
the method to the equipment available in most pharmaceutical

laboratories. The maceration should be caI:ried out in a heavy
walled bottle (a magnesium citrate bottle serves very well),
since the volatile solvents generate considerable pressure when
warmed to 40C.
2. "Cool the mixture, decant or filter off 100 cc. of the clear solution, and
evaporate this rapidly in a tared beaker or wide-necked flask to a volume
of about 5 cc. Add 5 cc. of chloroform to the residue and set it aside in a
moderately warm place."
The evaporation of the volatile solvents should be performed

as rapidly as possible, because cantharidin begins to sublime at
86C. and appreciable amounts are driven off with the solvent
during slow evaporation. When chloroform is added and the
mixture is allowed to stand in a warm place, the cantharidin
crystallizes out upon evaporation of the chloroformic solvent.
3. "When the solvent has all evaporated, add to the crystals 10 cc. of
a mixture of equal volumes of dehydrated alcohol and purified petroleum
benzin, which has previously been saturated with pure cantharidin, allow
the mixture to stand fifteen minutes, and then decant the liquid through
a pledget of purified cotton. Wash the crystals with successive portions
of a saturated solution of cantharidin, similar to that dtr;cted above, until
free from fat and coloring matter, and pass the washings througl). the same
pledget of purified cotton."
\
Cantharidin is only slightly soluble in the purified petroleum

benzin and dehydrated alcohol mixture. The use of a mixture
of these solvents previously saturated with cantharidin makes it
possible to remove the fat, coloring matter, etc., with which the
crystals are contaminated without dissolving any of the crystals.
Successive 2 to 3 cc. portions of the above wash mixture serve
most efficiently to remove the impurities adhering to the crystals.
4. "Then wash the cotton with a very small quantity of warm chloroform
to dissolve any adhering crystals, collect the chloroform in the tared flask
or beaker containing the washed crystals, evaporate the solvent with the
aid of a current of air, dry the crystals at 60C. for one-half hour, and weigh.
The resulting weight will be the amount of cantharidin obtained from
10 Gm. of Cantharides."
Negligible quantities of the cantharidin are volatilized when

it is dried for one-half hr. at 60C. The residue, which consists of
cantharidin and cantharidic acid in variable amounts, is con-
ASSAYS INVOLVING THE USE OF IMMISCIBLE SOLVENTS
439
sidered to be cantharidin. The weight of the residue corresponds

to the cantharidin contained in 10 Gm. of the sample assayed.
The U .-s.P. requires that cantharides yield not less than 0.6
per cent of cantharidin when assayed as directed above. Calculate the percentage of cantharidin contained in the sample
assayed and compare the results with the official requirement.
1. Why is hydrochlorfc acid added in the initial extraction of the
cantharides?
2. Why should the residue be dried at a low temperature?
3. Why is the maceration conducted with a warm solvent in a pressure
bottle?
Exercise 132
Object.-Assay of Oleoresin of Aspidium.

Materials Required.-About 3 Gm. of oleoresin of aspidium.
150 cc. of ether.
3 Gm. of barium hydroxide.
Procedure.-l. "Warm the Oleoresin on a water bath and stir until it is
thoroughly mixed. Transfer about 3 Gm. of it, accurately weighed, to a
250-cc. flask, dissolve it in 40 cc. of ether, add 75 cc. of a 3 per cent aqueous
solution of barium hydroxide, and shake the mixture vigorously for five
minutes. Transfer the mixture to a separator, allow the liquids to separate
completely, and draw off and filter the barium hydroxide layer. Rinse the
250-cc. flask with two 25-cc. portions of a 3 per cent aqueous barium hydroxide solution. Mter each rinsing, transfer the barium hydroxide solution
to the separator, shake the mixture for one minute, allow the liquids to
separate completely, and draw off and filter the barium hydroxide layer."
Oleoresin of aspidium usually deposits granular, crystalline

filicic acid or filicin upon standing. It is warmed to render it
more fluid so that the precipitate may be uniformly distributed
throughout the sample. The filmarone originally present, in
the crude drug, aspidium, is converted largely into filicic acid and
its amorphous modification, filicin, during the preparation of the
oleoresin. The barium hydroxide neutralizes the filma'l'one,
filicic acid, and filicin, forming the water-soluble barium compounds which dissolve in the aqueous layer of the immiscible
solvents. The resin, fat, coloring matter, etc., remain dissolved
in the ethereal layer.
440
2. "Transfer the combined filtered barium hydroxide solutions to a

separator, render distinctly acid to litmus paper by the addition of hydrochloric acid, and extract with three successive portions of 30 cc., 20 cc., and
15 cc. of ether."
The filicic acid and filicin are liberated from their barium salts
and dissolved in the ether, leaving barium chloride in the aqueous
solution.
3. "Filter the combined ethereal solutions, wash the filter with ether,
evaporate, and dry the residue to constant weight at 100C. This residue
is calculated as crude filicin and its weight should be not less than 24 per
cent of the weight of Oleoresin taken for the assay."
The residue consists mostly of filicin and small amounts of

filicic acid.
Calculate the percentage of crude filicin contained in the
oleoresin of aspidium, assayed, and compare the result with the
official requirements.
1. Why should the oleoresin be warmed and mixed well before the sample
is taken?
2. Why is the sample of oleoresin dissolved in ethe;? What function
does the ether serve?
\
3. Look up the assay method for aspidium in the U.S.P. and explain
briefly how this crude drug is assayed.
Exercise 133
Object.-Assay of Jalap.
Materials Required.-lO Gm. of jalap in No. 60 powder_
20 cc. of 1 per cent hydrochloric acid.
120 cc. of alcohol.
Procedure.-l. Place 10 Gm. of jalap, in fine powder and accurately
weighed, with about 60 cc. of a mixture of 9 volumes of alcohol and 1 volume
of water, in a flask provided with a reflux tube or condenser, and digest the
mixture on a steam bath during Y2 hr. Transfer the warm mixture to a
small-percolator, allow it to drain, press the marc down gently, and percolate
with the warm alcohol-water mixture until 100 cc. of percolate, when cooled,
is obtained, and mix thoroughly.
The jalap is digested and percolated with alcohol to extract

t.he resin completely.
441
2. Pipette 20 cc. of this percolate into a separator, add 40 cc. of chloroform

and 10 cc. of approximately 1 per cent hydrochloric acid, and shake for
2 rn'in. Allow the mixture to separate, draw off the chloroformic layer, and
extract the aqueous layer twice more, using 15 cc. of a mixture of 2 volumes
of chloroform and 1 volume of alcohol each time. Shake the combined
chloroformic solutions with 10 cc. of 1 per cent hydrochloric acid, again
separate, and wash this acid liquid twice with 15 cc. portions of chloroform-alcohol.
The resin of jalap is insoluble in aqueous acid solution. It

dissolves in the alcohol-chloroform layer leaving non-resinous
constituents in the aqueous layer.
3. Pass the combined chloroformic extractions through filter paper moistened with the chloroform-alcohol mixture, wash the filter with the chloroform-alcohol mixture, and evaporate the filtrate to dryness; add 2 cc. of
alcohol and again evaporate. Dry the residue to constant weight at 100C.
The weight obtained represents the yield of total resins from 2 Gm. of
jalap.
The lower aqueous layer containing small amounts of sugars,

gums, coloring matter, etc., is drawn off and discarded. The
alcohol-chloroform solution when evaporated to constant weight
leaves a residue which consists of practically pure resin of jalap.
Ipomoea and podophyllum are also assayed by analogous
methods, since the solubility properties of the resins which they
contain are similar to those of resin of jalap.
Exercise 134
Object.-Assay of Tablets of Phenobarbital.

Materials Required.-20 tablets of phenobarbital.
10 cc. of a 2 per cent solution of NaOH saturated with NaCl.
About 50 cc. of ether.
About 75 cc. of chloroform.
Procedure.-l. "Weigh not less than 20 of the Tablets, and reduce them
to a fine powder without an appreciable loss. Transfer an aliquot portion,
equivalent to about 0.3 Gm. of phenobarbital, to a separatory funnel, and
dissolve it, as far as possible, in 10 cc. or an alkaline salt solution made by
saturating an aqueous 2 per cent sodium hydroxide solution with sodium
chloride. Wash the solution with two 15 cc. portions of ether and discard
the washings."
The phenobarbital reacts with the NaOH to form soluble

phenobarbital. When the solution is shaken with portions of
442
ether, organic matter, such as petrolatum used as a lubricant in

the preparation of the tablets, dissolves in the ether but the sdlubIe phenobarbital being insoluble in ether remains in the aqueous
layer.
2. "Add 2 cc. of hydrochloric acid and 5 cc. of distilled water, and completely extract the phenobarbital with a solvent composed of two volumes
of alcohol, one volume of ether, and seven volumes of chloroform. Wash
the extract with 5 cc. of distilled water, acidified with a drop of hydrochloric
acid. Filter the extract through a pledget of cotton into a tared beak~,
TABLE 'LXVIII.-OFFICIAL
NON-ALKALOIDAL
SUBSTANCES
ASSAYED
BY
METHODS EMPLOYING IMMISCIBLE SOLVENTS
Substance
U.S.P.
Aspidium ........................... .
Aspidium, oleoresin of ................ .
Cantharides. . . . . . . . . . . . . . . . .. ... . ..
Podophyllum ................. .
Soluble barbital. ............. .
Soluble phenobarbital. ..
N.F.
Acetophenetidin, tablets of. .......... .
Acetophenetidin and phenyl salicylate,
tablets of. .. .. . . . . .. . ........ .
Aminopyrine, elixir of . ......... .
Aminopyrine, tablets of. . . .. . ..
Barbi tal, elixir of. . . .. ...... .
Barbital, tablets of, more than 0.07 Gm ..
0.07 Gm. orle8s .......... .
Ipomoea .................. .
Jalap...... . . . . . . . . . . . . . . . . . .
Jalap, fluidextract of. .. . . ... ..
Jalap, tincture of ...... , ... .
Phenobarbital, elixir of ........ .
Phenobarbital, tablets of
more than 0.07 Gm .. : .......... .
0.07 Gm. or less ....
Soluble barbital, tablets of
more than 0.07 Gm ............... .
0.07 Gm. or less .................. .
Soluble phenobarbital, tablets of
more than 0.07 Gm ...
0.07 Gm. orless ...
Amount
used,
Gm. or
cc.
0.5
Crude filicin = 1.5

Crude filicin = 24
Cantharidin = 0.6
Resin = ~
Barbital = 88 to 90
Phenobarbital = 90.4 to 91.4
0.3
Acetophenetidin 1= 92.5 to 107. 5b
5
I"
10
0.3"
0.3
10
10
2
10
25
Aminopyrine = 3.7 to 4.3W/V

Aminopyrine = 92.5 to 107. 5b
Barbital = 3.2 to 3.8W/V
Barbital = 92.5 to 107. 5 b
Barbital = 91 to 109b
Total resins = 15
Total resins = 9
Resin = 8.5 to 9.5WIV
Resin = 1. 7 to 1. 9W IV
Phenobarbital = 0.38 to 0.42W/V
125
3
15
10
0.3
Phenobarbital = 92.5 to 107. 5 b

Phenobarbital = 91 to 109b
0.3
0.3
Barbital = 92.54,0 107. SbBarbital = 91 to 109 b
0.3 a
0.3 a
Phenobarbital = 92.5 to 107. 5b

Phenobarbital = 91 to 109b
0.3

443
and wash the cotton with small portions of the solvent. Evaporate the
combined filtrate and washings on a water bath with the aid of a current of
air, and dry the residue of phenobarbital to constant weight at a temperature
not exceeding 100C."
The Hel liberates the phenobarbital from the soluble" phenobarbital and the-liberated phenobarbital is dissolved and extracted
upon shaking with successive portions of the mixed organic
solvents_
1. Calculate the amount of phenobarbital in each of the tablets.
2. Docs thc amount of phenobarbital found fall within the tolerance
limits established for these tablets in the National Formulary?
3. Consult the U.S. Pharmacopoeia for the formulae of phenobarbital
and soluble phenobarbital and write equations for the reactions that occur
in the assay using graphic for~ulae.
CHAPTER XXVIII
ASSAY OF ENZYME-CONTAINING SUBSTANCES
No methods are known whereby the quantity of enzymes in a
substance can be accurately determined. ,Consequently, the
assay of enzyme-containing substances is based on observations
of their power to produce changes in other substances. In
other words, the activity of the enzymes is measured. Thus,
pepsin is evaluated on the basis of the quantity of egg albumen
which it will digest, and extract of malt is assayed for its power to
convert starch into soluble carbohydrates under definite conditions. Any condition which tends to increase or decrease the
activity of the en'zymes manifests itself in the assay results as
an increase or decrease in the quantity of enzyme;; present.
Numerous factors tend to influence the activity of enzymes.
Some of the more important of these factors are as foqows:
L Practically all enzymes are destroyed when they are heated
to 100C. Many 'of them are gradually inactivated at comparatively low temperatures (35 to 40C.), and many of them
undergo a slow loss of activity at room temperature and even at
OC.
2. Enzymes have an optimum temperature of action; that is,
for each enzyme there is a definite range of temperature at
which it exhibits its maximum activity under definite conditions.
The optimum temperature for most enzymes lies between 30 and
45C.
3. The activity of the enzymes is affected by the media in
which they react; e.g., pepsin is most active in acid media, and
pancreatin is most active in alkaline media.
4. The enzymes are inactivated by many chemical agents,
such as alcohol, salts of the heavy metals, tannin, and phosphotungstic acid, which precipitate the enzymes from solution.
Because the activity of enzymes is influenced by so many
factors, it is necessary to adhere strictly to the procedures out444
445
lined in the various assay processes in order to secure concordant

results.
Exercise 135
Object.-Assay of
P~psin.
Materials Required.-O.1 Gm. of pepsin.

0.1 Gm. of U.S.P. reference pepsin. The U.S.P. reference pepsin consists
of a powdered pepsin, carefully selected, dried, and packaged and having a
digestive power of 3,000 times its weight of egg albumen.
Normal hydrochloric acid.
A hen's egg.
A No. 40 sieve.
A conical measure.
Procedure.-l. "Mix 35 cc. of normal hydrochloric acid with 385 CC. of
distilled water. Dissolve 0.1 Gm. of Pepsin in 150 co. of this dilute acid.
Likewise dissolve 0.1 Gm. of Reference Pepsin in another portion of 150 cc.
of this dilute acid."
When 35 cc. of normal Hel is diluted to 420 cc., the concentration of acid in the solution is reduced to approximately 0.3 per
cent. This is about the normal acidity of the stomach. Investigation has shown that pepsin is most active as an albumen digestive in media containing this concentration of acid. Other
strongly dissociated acids which supply the same hydrogen ion
concentration as 0.3 per cent Hel might be used, since the digestive activity of the pepsin is directly related to the hydrogen ion
concentration of the reaction mixture.
2. "Immerse one or more hen's eggs in boiling water during fifteen
minutes. Cool them rapidly to room temperature by immersion in cold
water, remove the shell and pellicle and all of the yolk and at once rub the
albumen through a clean, dry, No. 40 sieve, rejecting the first portion that
passes through the sieve. Place 10 Gm. of the succeeding well-mixed portion in each of three wide-mouth bottles of about 100-cc. capacity."
The egg is immersed in boiling water for 15 min. to thoroughly

coagulate the albumen. The albumen is coagulated so that its
digestion by the pepsin may be bet~er observed. The coagulated
albumen is passed through a sieve to break it into small particles
so that a greater surface area will be exposed to the digestive
action of the pepsin enzymes. The first portion of the albumen
that passes through the sieve is rejected, since it may contain
446
QUANTITATIV,E PHARMACEUTICAL CHEMISTRY
dust particles and traces of metallic compounds from the materials of the sieve.
3. "Immediately add 35 cc. of the dilute acid at one time or in portions
and, by suitable means, disintegrate thoroughly the particles of albumen.
Place the bottles in a water bath at 52C. After the contents of the bottles
have reached that temperature, add exactly 5 cc. of the acidulated solution
of Pepsin to one bottle, 4.30 oc. of the same solution and 0.70 co. of the dilute
acid to another bottle, and exactly 5 cc. of the acidulated solution of Reference Pepsin to the third bottle. At onoe stopper the bottles s'ecurely, invert
them three times, and maintain them at a temperature of 52C. for two and
one-half hours, agitating the contents equally every ten minutes by inverting
the bottles once."
The acid should be added at once, because the

albumen loses moisture on exposure to the air and
becomes tough and hard. The mixture is maintained at 52C., since pepsin exerts its optimum
activity at this temperature. Exactly 5 cc. of the
pepsin solution being assayed and of the solution
of reference pepsin is added, because this quantity
FIG. 67.- introduces 1 part of pepsin intp~the mixture to each
Sedimenta3,000 parts of albumen. Thus 0.1 Gm. of pepsin
tion cone.
in 150 cc. gives 0.1: 150: : X: 5. X = tl.003333 Gm.
pepsin in 5 cc . of the solution. 0.0033 X 3,000 = 9.999 (10
Gm.), the weight of albumen used.
4. "Remove the bottles from the bath, pour the contents into 100-cc.,
conically-shaped measuring vessels, Fig. 67, having diameters not exceeding
1 cm. at the bottom, and graduated from the tip to the 1.0-co. mark in
0.05-00: divisions and from the 1.0-co. to the 5.0-cc. mark in O.l-oc. divisions,
and having the internal taper as nearly identical as possible. Transfer the
undigested egg albumen which adheres to the sides of the bottles to the
respective measuring vessels with the aid of small portions of distilled water
until 50 co. has been used for each. Mix the contents of eaoh measuring
vessel and allow them to stand for thirty minutes. The volume of the
undissolved albumen in the measuring vessel corresponding to 5.0 cc. of the
solution of Pepsin being assayed shall not be more than the volume of
the undissolved albumen in the measuring vessel corresponding to 5.0 cc.
of the Reference Pepsin solution, and the volume of the undissolved albumen
in the measuring vessel corresponding to 4.30 cc. of the solution of Pepsin
being assayed, shall not be less than the volume of the undissolved albumen
in the measuring vessel corresponding to 5.0 oc. of the Reference Pepsin
solution."
447
If the volume of egg albumen remaining undissolved from the

sample containing 5.0 cc. of the solution of pepsin being assayed
is not greater than that remaining from the sample containing
the solution of reference pepsin, it is evident that the pepsin
being assayed must be at least as active as the reference .pepsin
and that it must conform to the minimum official requirement.
If the volume of egg albumen remaining undissolved from the
sampie containing 4.3 cc. of the solution of pepsin being assayed
is not less than that remaining from the sample containing the
reference pepsin, it is evident that the pepsin being assayed is
a pepsin which will cause the solution of 3,500 times its weight
of egg albumen. 4.3: 5: : 3,000: X. X = 3,500. The relative
proteolytic power of pepsin, stronger or weaker than that just
described, may be determined by ascertaining through repeated
trials the quantity of the pepsin solution made as directed in the
assay, required to digest under the prescribed conditions, 10 Gm.
of boiled and disintegrated egg albumen.
1. Name several factors that affect the activity of pepsin.
2. Why is the proteolytic activity of pepsin determined in acid solution?
What effect would the use of a neutral or alkaline medium have on the digestive power of pepsin?
3. Oould proteins other than egg albumen be used to estimate the proteolytic activity of pepsin? Give examples.
4. What advantages does egg albumen possess as a source of protein for
this assay?
Exercise 136
Object.-Assay of Pancreatin for Starch Digestive Power.

Materials Required.-5 Gm. of powdered potato starch.
0.3 Gm. of pancreatin.
0.2 cc. of 0.1 N iodine solution.
Procedure.-l. "Determine the percentage of moisture in potato starch
by drying about 0.5 Gm., accurately weighed, at 1200., for four hours.
Thoroughly mix a quantity of the starch, equivalent to 3.75 Gm. of dry
starch, with 10 cc. of cold distilled water. Add the mixture with constant
stirring to 75 cc. of distilled water, previously heated to from ,50 to 600.,
contained in a tared, 250-cc. beaker. Rinse the remaining starch into the
beaker with 10 ee. of distilled water, heat the mixture to boiling, and boil it
gently, with constant stirring, for five minutes, or until a translucent, uniform paste is obtained."
448
The percentage of moisture remaining in the stch is determined so that a sample equivalent to 3.75 Gm. of dry starch
can be used for digestion. Thus, if the sample dried at 500.
is found to contain 10 per cent of moisture, each gram of the
sample is 'equivalent to 0.9 Gm. of dry starch. Then 3.75/0.9 =
4.1666 Gm. starch equivalent to 3.75 Gm. of dry starch.
The starch is mixed with cold water to obtain a paste so that it
will not form lumpy masses when the hot water is added. When
this paste is added to warm water and the resulting mixture is
boiled, the starch grains burst, liberating their contents which
dissolve in the hot water to form a more or less mucilaginous
mass. Until the starch grains are broken down in this way,
they remain insoluble and are not acted upon by pancreatin.
2. "Add enough distilled water to make the mixture weigh 100 Gm.,
cool the paste to 40C., and place the beaker on a water bath maintained
at 40C. Suspend 0.15 Gm. of Pancreatin in 5 cc. of distilled water and add
the suspension to the starch paste, mixing it well by pouring the mixture
from beaker to beaker for thirty seconds, noting the time when the Pancreatin suspension was added to the starch. Maintain the mixture at a
temperature of 40C. for exactly five minutes."
The water lost by evaporation is replaced so that a definite

concentration of starch will be present during the digestion with
the pancreatin. The solution of pancreatin must be prepared
immediately before it is added to the starch solution, because
amylopsin, the starch-splitting enzyme of pancreatin, becomes.
inactivated quite rapidly in aqueous solutions. The mixture is
maintained at 40 0 e. for 5 min., since 40 0 e. is the optimum
temperature for amylopsin. The regulation of the temperature
is very important in this assay, since a variation of 1e. throughout the period of digestion has been found to cause a difference
of about 10 per cent in the activity of the amylopsin of pancreatin.
The amylopsin converts the starch into soluble carbohydrates,
chiefly maltose and isomaltose.
3. "At once add 0.1 cc. of this mixture to a previously made mixture of
0.2 cc. of tenth-normal iodine and 60 cc. of distilled water at a temperature
of from 23 to 25C.: no blue, red or violet color is produced."
The presence of starch and primary hydrolytic products of

starch is indicated by the development of a blue, red, or violet
color.
449
Pancreatin must be capable of converting at least twenty-five

times its weight of starch into soluble carbohydrates in order
to meet the pharmacopoeial requirements. Since 0.15 Gm. of
pancreatin is employed to digest 3.75 Gm. of starch, each Gill. of
pancreatin used corresponds to 3.75/0.15 = 25 Gm. starch. The
relative amy10ptic activity of a sample of pancreatin can be
determined by repeated trials on varying quantities of starch.
Extract of malt of the U.S.P. is also assayed for its capacity to
convert starch into soluble carbohydrates which do not give a
color with iodine solution.
1. Define the terms proteolytic enzyme and amylolytic enzyme.
2. Why is the starch solution boiled before the digestion with pancreatin?
3. Look up the hydrolytic products of starch and explain what products
might give a red or violet color with i?dine?
Exercise 137
Object.-Assay of Pancreatin for Casein Dige'stive Power.

Materials Required.-D.l Gm. of casein.
1 cc. of 0.1 N sodium hydroxide.
01. Gm. of pancreatin.
1 cc. of glacial acetic acid.
10 cc. of alcohol.
Procedure.-1. "Place O:I'Gm. of finely powdered casein in a 50 cc. volumetric flask, add 30 cc. of distilled water, and shake well to bring the casein
into suspension. Add exactly 1 cc. of tenth-normal sodium hydroxide, and
heat the mixture at 40C. until the casein is completely dissolved, which
should not require more than thirty minutes. Cool, add sufficient distilled
'water to make 50 cc., and mix well."
Casein is a complex compound, or mixture of compounds,

which yields amino acids upon hydrolysis. Consequently, it is
soluble in alkalies. The casein is dissolved in alkali, in this case,
because trypsin, the proteolytic enzyme contained in pancreatin,
functions best in alkaline wedia. Casein which requires more
than 30 min. to dissolve is usually of poor quality, containing an
excess of fat.
2. "Dissolve 0.1 Gm. of Pancreatin in 500 cc. of distilled water. Mix 1 cc.
of glacial acetic acid with 9 cc. of distilled water and 10 cc. of alcohol. Place
5 cc. of the casein solution in a test tube, add to it 2 cc. of the well-shaken
Pancreatin solution and 3 cc. of distilled water, and mix by gentle agitation.
450
QUANTITATIVE PHARMACEU'rICAL CHEMISTRY
Immediately immerse the test tube in a water bath at 400., and 'keep it
at this temperature for one hour. Then remove from the bath, and add 3
drops of the acetic acid mixture. No precipitate is produced."
The concentrations of the solutions of casein and pancreatin

are such that when they are mixed there will be 25 parts of casein
present for each part of pancreatin. Thus, in 2 cc. of the pancreatin solution there is 2 X
~O~ =
0.0004 Gm. pancreatin, and
in 5 cc. of casein solution there is 5 X
~.~ =
0.01 Gm. casein;
0.0004 X 25 = 0.0100.
The mixture is maintained at 40C., the optimum temperature
for trypsin, for 1 hr. to allow the proteolytic action to proceed.
The casein is digested by the pancreatic enzymes with the
formation of soluble proteoses and possibly small amounts of
amino acids. When the alcoholic acetic acid solution is added,
the alkali is neutralized and any undigested casein is precipitated, since casein is insoluble in alcohol and in acetic acid.
The proteoses and other products of casein digestion, being
soluble in alcohol and acetic acid, are not precipitated. Consequently, if a precipitate is formed, it indicates that the proteolytic activity of the pancreatin is below stanQ.ard; that is,
that 1 part of the pancreatin is not capable of digesting 25 parts
of casein.
1. Why is the protein digestive capacity of pancreatin determined in an
alkaline medium?
2. Which of the enzymes of pancreatin take part in protein digestion?
3. Define the term optimum temperature as applied to enzymes.
4. How might the relative proteolytic power of a sample of pancreatin
stronger or weaker than the official standard be determined?
Exercise 138
Object.-Assay of Rennin.
Materials Required.--o.l Gm. of rennin.
0.1 Gm. N.F. reference rennin (a carefully preserved, stable, powdered
rennin that has been repeatedly tested for a number of years so that its
stability and its standard are definitely established. It coagulates approximately but not less than 25,000 times its weight of fresh cow's milk).
100 cc. of cow's milk.
451
Procedure.-l. "Mix 0.1 Gm. of Rennin with 50 cc. of distilled water by

gentle stirring, and allow the mixture to stand exactly 15 minutes. Likewise, mix 0.1 Gm. of Reference Rennin (see page 450) with 50 cc. of distilled
water by gentle stirring, and allow the mixture to stand for exactly 15
minutes."
When rennin is added to water, it is taken up slowly, forming a

somewhat opalescent mixture. If shaken vigorously, the rennin
tends to clump into large particles which separate out. When
the mixture is stirred gently and allowed to stand, however, the
rennin gradually absorbs water and dissolves.
2. "Place 50 cc. of well-mixed cow's milk in each of two glass vessels
about 12 cm. high and 4.5 cm. in diameter. Warm the milk rapidly to a
temperature of 430., and maintain it on a water bath at this temperature.
Add 1 cc. of the rennin solution to the milk in one vessel, and 1 cc. of the
Reference Rennin solution to the milk in the other vessel, noting the time
when each solution is added. Stir each milk mixture slowly for 10 seconds
immediately after the solution is added. Note the time which elapses until
the milk thickens, as shown by a distinctly convex surface when the vessel
is tipped to an angle of about 45."
The rennin solution is added to the milk, and the mixture is

maintained at 43C., because rennin exhibits its optimum
activity at that temperature. The mixture is stirred to obtain
an equal distribution of the rennin throughout the milk. The
thickening of the milk is caused by the conversion of the soluble
casein in the milk into soluble paracasein and insoluble calcium
paracasein.
3. "The rennin solution should coagulate in not less than 90 per cent and
not more than 110 per cent of the time required by the Reference Rennin
solution.
"NOTE: Milk varies in its coagulability. Different lots of milk, or the
same distributor's milk obtained on successive days, will be found to vary in
coagulation time with the Reference Rennin."
.
The time required for the coagulation of milk by rennin has

been found to be inversely proportional to the amount of rennin
present. This relationship may be formulated as follows:
eXT = K where C represents the concentration of rennin
and T represents the time during which it acts on the milk and K
is a constant. It is obvious that when the amount of rennin
used is reduced, the time required for the coagulation of a given
amount of milk will be increased.
452
The National Formulary specifies that rennin should be

capable of coagulating twenty-five thousand times its weight of
fresh milk. In the assay 0.002 Gm. of rennin contained in 1 cc.
is used to coagulate 50 cc. of milk. Therefore, 1 Gm. of rennin
is used for each 50/0.002 = 25,000 cc. milk.
The relative milk-coagulating power of rennin stronger or
weaker than the specified strength may be determined by
repeated trials, using larger or smaller quantities of rennin.
1. What substances tend to accelerate the activity of rennin?
2. If a 50 cc. sample of milk is found to have an acidity of 0.175 per cent,
how much normal NaOH solution would be required to reduce the acidity
to 0.145 per cent?
3. Milk usually has a specific gravity of from 1.029 to 1.034. The
National Formulary directs that rennin should be capable of coagulating
twenty-five thousand times its weight of fresh milk. In the assay, howev~r,
a volume of 50 cc. of milk is employed for each 0.002 Gm. of rennin. If the
milk used has a specific gravity of L030, what should be the weight-to-weight
ratio of the milk coagulated to the rennin?
TABLE LXIX.-OFFICIAL SUBSTANCES ASSAYED FOR ENZYME ACTIVITY
Substrate
Substance
Official requirement
\
U.S.P.
Malt, extract of. . . . . . . .. Potato starch
Pancreatin ............ , Potato starch
Casein
Pepsin. . . . . . . . . . . . . . . .. Egg albumen
Converts 5 >< its wt. of starch

into soluble sugars.
Converts 25 X its wt. of starch
into soluble carbohydrates.
Converts 25 'x its wt. of casein
into soluble proteoses.
Digests 3,000 to 3,500 X its
wt. of egg 4lbumen.
N.F.
Pepsin, compound elixir Egg albumen
of
Pepsin, elixir of. . . . . . . .. Egg albumen
Pepsin, elixir of and ren- Egg albumen
run
Pepsin, glycerlte of. . . . .. Egg albumen

Rennin. . . . . . . . . . . . . . .. Milk
Rennin, elixir of pepsin Milk
and
100 cc. = 1.75 Gm.

ence pepsin.
100 cc. = 1.75 Gm.
ence pepsin.
100 cc. = 2.25 Gm.
ence pepsin.
100 cc. = 8.75 Gm.
ence pepsin.
Coagulates 25,000 X
100 cc. = 1.75 Gm.
ence rennin.
of referof referof referof referits wt.

of refer-
453
APPENDIX
LOGARITHMS OF NUMBERS
Proportional parts
Natural
numbers
9
11213141516171819
12
13
14
10
0000 0043 0086 0128 0170 0212 0253 0294 0334 0374
0414 0453 0492 0531 0569 0607 0645 0682 0719 0755
0792 0828 0864 0899 0934 0969 1004 1038 1072 1106
1139 1173 1206 1239 1271 1303 1335 1367 1399 1430
1461 1492 1523 1553 1584 1614 1644 167a 1703 1732
4
4
3
3
3
8 12 17 21 25 29 3337
811 15 19 23 26 30 34
7 10 14 17 21 24 2831
6 10 13 16 19 23 2629
6 912 15 1821 24 27
15
16
17
18
19
1761 1790 1818 1847 1875 1903 1931 1959 1987 2014
2041 2068 2095 2122 2148 2175 2201 2227 2253 2279
2304 2330 2355 2380 2405 2430 2455 2480 2504 2529
2553 2577 2601 2625 2648 2672 2695 2718 2742 2765
2788 2810 2833 2856 2878 2900 2923 2940 2967 2989
3
3
2
2
2
6
5
5
5
4
811 1417 20 2225

811 1316 1821 24
710 1215 1720 22
7 9 1214 1619 21
7 9 1113 1618 20
20
21
22
3010 3032 3054 3075 3096 3118 3139 3160 3181 3201
3222 3243 3263 3284 3304 3324 3345 3365 3385 3404
3424 3444 3464 3483 3502 3522 3541 3560 3579 3598
3617 3636 3655 3674 3692 3711 3729 3747 3766 3784
3802 3820 3838 3856 3874 3892 3909 3927 3945 3962
2
2
2
2
2
4
4
4
4
4
6 811 1315 1719

6 810 1214 1618
6 810 1214 1517
6 7 9 1113 1517
5 7 9 1112 141t!
25
26
27
28
3979 3997 4014 4031 4048 4065 4082 4099 4116 4133
4150 4166 4183 4200 4216 4232 4249 4265 4281 4298
4314 4330 4346 4362 4378 4393 4409 4425 4440 4456
4472 4487 4502 4518 4533 4548 4564 4579 4594 4609
4624 4639 4654 4669 4683 4698 4713 4728 4742 4757
2
2
2
2
1
3
3
3
3
3
5
5
5
4
80
4771 4786 4800 4814 4829 4843 4857 4871 4886 4900
4914 4928 4942 4955 4969 4983 4997 5011 5024 5038
5051 5065 5079 5092 5105 5119 5132 5145 5159 5172
5185 5198 5211 5224 5237 5250 5263 5276 5289 5302
5315 5328 5340 5353 5366 5378 5391 5403 5416 5428
1
1
1
1
1
3
3
3
3
3
4
.4
4
4
11
23
24
29
81
82
83
84
5502 5514 5527 5539 5551

5623 5635 5647 5658 5670
5740 5752 5763 5775 5786
5855 5866 5877 5888 5899
5966 5977 5988 5999 6010
5 7 910 1214 18
7
6
6
6
810 1113 15
8 9 1113 14
8 9 1112 14
7 9 1012 13
4 6 7 910 1113
6 7 810 1112
5 7 8 9 1112
5 6 8 9 1012
5 6 8 9 1011
1 2 4
1 2 4
1 2 3
1 2 3
1 2 3
39
5441 5453 5465 5478 5490

5563 5575 5587 5599 5611
5682 5694 5705 5717 5729
5798 5809 5821 5832 5843
5911 5922 5933 5944 5955
40
41
42
43
44
6021 6031 6042 6053 6064 6075 6085 6096 6107 6117
6128 6138 6149 6160 6170 6180 6191 6201 6212 6222
6232 6243 6253 6263 6274 6284 6294 6304 6314 6325
6335 6345 6355 6365 6375 6385 6395 6405 6415 6425
6435 6444 6454 6464 64_74 6484 6493 6503 6513 6522
1
1
1
1
1
2
2
2
2
2
3 4 5 6
3 4 5 6
3 4 5 6
3 4 5 6
3 4 5 6
45
46
47
48
6532 6542 6551 656i 6571 6580 6590 6599 6609

6628 6637 6646 6656 6665 6675 6684 6693 6702
6721 6730 6739 6749 6758 6767 6776 6785 6794 6803
6812 6821 6830 6839 6848 6857 6866 6875 6884 6893
6902 6911 6920 6928 6937 6946 6955 6964 6972 6981
~m
1
1
1
1
1
2
2
2
2
2
3 4
3 4
3 4
3 4
5
5
5
4
3 4 4
50
6990 6998 7007 7016 7024 7033 7042 7050 7059 7067 1
70711 7084 7093 7101 7110 7118 7126 7135 7143 7152 1
7160 7168 7177 7185 71937202 7210 7218 7226 7235 1
7243 7251 7259 7267 72757284 7292 7300 7308 731611
7324 7332 7340 7348 73567364 7372 7380 7388 7396 1
2
2
2
2
2
3 3 4 Ii 6 7 8
3 3 4 5 6 7 8
35
36
87
88
49
51
52
53
54
5 6 7 9 1011
5
5
5
4
6
6
6
5
7
7
7
7
8 1011
8 910
8 910
8 910
8
7
7
7
7
910
8 9
8 9
8 9
8 9
6 7 8 9
6 7 7 8
5 6 7 8
.5 6 7 8
5 6 7 8
2 3 4 5 6 7 7
2 3 4 /; 6 6 7
2 3 4 Jj 6 6 7
* See page 13 for example. illustrating the us","of logarithmic table. in calculations.
454

'LOGARITHMS OF NUMBERS.-(Continued)
Proportional parts
Natural
numbers
9
11213141516171810
55
56
57
68
59
7404 7412 7419 7427 7435 7443 7451 7459 7466 7474
7482 7490 7497 7505 7513 7520 7528 7536 7543 7551
7559 7566 7574 7582 7589 7597 7604 7612 7619 7627
7634 7642 7649 7657 7664 7672 7679 7686 7694 7701
7709 7716 7723 77a1 7738 7745 7752 7760 7767 7774
1
1
1
1
1
2
2
2
1
1
2
2
2
2
2
3
3
3
3
3
4
4
4
4
4
5
5
5
5
5
6
6
6
6
6
7
7
7
7
7
60
61
62
63
64
7782 7789 7796 7803 7810 7818 7825 7832 7839 7846
7853 7860 7868 7875 7882 7889 7896 7903 7910 791~
7924 7931 7938 7945 7952 7959 7966 7973 7980 798
7993 8000 8007 8014 8021 8028 8035 8041 8048 8055
8062 8069 8075 8082 8089 8096 8102 8109 8116 8122
1
1
1
1
1
1 2
1 2
1 2
1 2
1 2
3
3
3
3
3
4 4 5
4 4 5
3 4 5
3 4 5
3 4 5
6
6
6
5
5
6
6
6
6
6
65
66
67
68
69
8129 8136 8142 8149 8156 8162 8169 8176 8182 8189
8195 8202 8209 8215 8222 8228 8235 8241 8248 8254
8261 8267 8274 8280 8287 8293 8299 8306 8312 8319
8325 8331 8338 8344 8351 8357 8363 8370 8376 8382
8388 8395 8401 8407 8414 8420 8426 8432 8439 8445
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
3
3
3
3
2
3
3
3
3
3
4
4
4
4
4
5
5
5
4
4
5
5
5
6
5
6
6
6
70
72
73
74
8451 8457 8463 8470 8476 8482 8488 8494 8500 8506
8513 8519 8525 8531 8537 8543 8549 8555 8561
8573 8579 8585 8591 8597 8603 8609 8615 8621 ~m
8633 8639 8645 8651 8657 8663 8669 8675 8681 8686
8692 8698 8704 8710 8716 8722 8727 8733 8739 8745
1
1
1
1
1
1
1
1
1
2
2
2
2
1 2
2
2
2
2
2
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5 6
5 /)
5 I)
75
76
77
78
79
8751 8756 8762 8768 8774 8779 8785 8791 8797 8802 1
8808 8814 8820 8825 8831 8837 8842 8848 8854 8859 1
8865 8871 8876 8882 8887 8893 8899 8904 8910 8915 1
8921 8927 8932 8938 8943 8949 8954 8960 8965 8971 1
8976 8982 8987 8993 8998 9004 900~ 9015 9020 9026 ,1
1
1
1
1
1
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
5
5
4
4
4
80
81
82
83
84
9031 9036 9042 9047 9053 9058 9063 9069 9074 9079
9085 9090 9096 9101 9106 9112 9117 9122 9128 9133
9138 9143 9149 9154 9159 9165 9170 9175 9180 9186
9191 9196 9201 9206 9212 9217 9222 9227 9232 9238
9243 9248 9253 9258 9263 9269 9274 92('9 9284 9289
1 1 212
1 1 2 2
1 1 2 2
1 1 2 2
1 1 2 2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
85
86
87
89
9~4 9299 9304 9309 9315 9320 9325 9330 9335 9340
9345 9350 9355 9360 9365 9370 9375 9380 9385 9390
9395 9400 9405 9410 9415 9420 9425 9430 9435 9440
9445 9450 9455 9460 9465 9469 9474 9479 9484 9489
9494 9499 9504 9509 91?13 9518 9523 9528 9533 9538
1
1
0
0
0
1
1
1
1
1
2
2
1
1
1
3
3
2
2
2
3
3
3
3
3
4
4
3
3
3
4
4
4
4
4
90
91
92.
93
94
9542 9547 95112 9557 9562 9566 9571 9576 9581 9586
9590 9595 9600 9605 9609 9614 9619 9624 9628 9633
9638 9643 9647 9652 9657 9661 9666 9671 9675 9680
9685 9689 9694 9699 9703 9708 9713 9717 9722 9727
9731 9736 9741 9745 97!i0 9754 9759 9763 9768 97;,?3
0
0
0
0
0
1
1
1
1
1
1 2 2
1 2 2
1 2 2
1 2 2
1 2 2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
95
96
97
98
99
0777 9782 9786 9791 9795 9800 9805 9809 9814 9818
9823 9827 9832 9836 9841 9845 981)0 9854 9859 9863
9868 9872 9877 9881 9886 9890 9894 9899 9903 9908
9912 9917 9921 9926 9930 9934 9939 9943 9948 9952
99b6 9961 9965 9969 9974 9978 9983 9987 9991 9996
0
0
0
0
0
1
1
1
1
1
1
1
1
1
1
2 2
2 2
2.2
2 2
2 2
3
3
3
3
3
3 4 4.
3 4 4
71
88
2
2
2
2
2
2
2
2
2
2
5
5
5
4
4
I)
I)
5 II
IS
5
5
.5
5
5
5
.5
5
4 5
IS
/)
4
4
4.
4.
4.
4.
4.
3 4 4.
3 4 4
3 3 4.
455
APPENDIX
ANTlLOGAlUTHMS
,Proportional pa.rts
Logarithms
1009
1033
1057
1081
1107
1012
1035
1059
1084
1109
1\2\3\415\6\7181~
a a
0
0
0
0
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
2
2 2 2
0
0
0
1
0
0
0
0
0
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2 2 2
2 2 2
.09
1122 1125 1127 1130 1132 1135 1138 1140 1143 1146
1148 1151 1153 1156 1159 1161 1164 1167 1169 1172
1175 1178 1180 1183 1186 1189 1191 1194 1197 1199
1202 1205 1208 1211 1213 1216 1219 1222 1225 1227
1230 1233 1236 1239 1242 1245 124'1 1250 1253 1256
.10
.11
.12
.13
.14
1259 1262
1288 1291
1318 1321
1349 1352
1380 1384
1274 1276 1279 1282 1285

1303 1306 1309 1312 1315
1334 1337 1340 1343 1346
1365 1368 1371 1374 1377
1396 ~400 1403 1406 1409
0
0
0
0
0
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
2
2
2
2
.15
.16
.17
.18
.19
1413
1445
1479
1514
1549
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
2 3 3
2 3 3
:I 3 3
2 3 a
3 3 3
.20
.21
.22
.23
.24
1585 1589 1592 1596

1622 1626 1629 1633
1660 1663 1667 1671
1698 1702 1706 1710
1738 1742 1746 1750
1600
1637
1675
1714
1754
1603 1607
1641 1644
1679 1683
1718 1722
1758 1762
.25
.26
.27
.28
.29
1778
1820
1862
1905
1950
1782 1786 1791

1824 1828 1832
1866 1871 1875
1910 1914 1919
1954 1959 1963
1795
1837
1879
1923
1968
.00
.01
.02
.03
.04
1000 1002
1023 1026
1047 1050
1072 1074
1096 1099
.05
.06
.07
.08
1005 1007
1028 1030
1052 1054
1076 1079
1102 1104
1265 1268
1294 1297
1324 1327
1355 1358
1387 1390
1416 1419 1422

1449 1452 1455
1483 1486 1489
1517 1521 1524
1552 1556 1560
1271
1300
1330
1361
1393
1014 1016
1038 1040
1062 1064
1086 1089
1112 1114
1426 1429 1432 1435

1459 1462 1466 1469
1493 1496 1500 1503
152~ 1531 1535 1538
156 1567 1570 1574
1019 1021
1042 1045
1067 1069
1091 1094
1117 1119
2 2 2
2 2 2
2 2 2
2 2 2
2 2 2
2 2 3
2 2
2
2
2
3
3
3
3
3
3
1439
1472
1507
1542
1578
1442
1476
1510
1545
1581
0
0
0
0
0
1
1
1
1
1
1611 1614
1648 1652
1687 1690
1726 1730
1766 1770
1618
1656
1694
1734
1774
0
0
0
0
0
1 1 1 2 2 3 3 3
1 1 2 2 2 3 3 3
1799 1803 1807 1811

1841 1845 1849 1854
1884 1888 1892 1897
1928 1932 1936 1941
1972 1977 1982 1986
1816
1858
1901
1945
1991
0
0
0
0
0
.30
.31
.32
.33
.34
1995 2000 2004 2009 2014 2018 2023 2028 2032 2037
2042 2046 2051 2056 2061 2065 2070 2075 2080 2084
2089 2094 2099 2104 2109 2113 2118 2123 2128 2133
2138 2143 2148 2153 2158 2163 2168 2173 2178 2183
2188 2193 2198 2203 2208 2213 2218 2223 2228 2234
0
0
0
0
1
1 1 2 2 3 3 4 4
1 1 2 2 3 3 4 4,
1 1 2 2 3 3 4 4
.35
.36
.37
.3S
.39
2239 2244 2249 2254 2259 2265 2270 2275 2280

2291 2296 2301 2307 2312 2317 2323 2328 2333
2344 2350 2355 2360 2366 2371 2377 2382 2388
2399 2404 2410 2415 2421 2427 2432 2438 2443
2455 2460 2466 2472 2477 2483 2489 2495 2500
2286
:1339
2393
2449
2506
1
1
1
1
1
1 2 2 3 3 4 4 Ii
.40
.41
.42
.43
.44
2512 2518 2523 2529 2535 2541 2547 2553 2559 2564
2570 2576 2582 2588 2594 2600 2606 2612 2618 2624
2630 2636 2642 2649 2655 2661 2667 2673 2679 2685
2692 2698 2704 2710 2716 2723 2729 2735 2742 2748
2754 2761 2767 2773 2780 2786 2798 2799 2805 2812
1
1
1
1
1
1
1
1
1
1
3
3
3
3
3
4
4
4
4
4
4 5 6
4 II 6
.45
.46
.47
.48
.49
2818 2825 2831 2838 2844 2851 2858 2864 2871 2877
2884 2891 2897 2904 2911 291.7 2924 2981 2938 2944
2951 2958 2965 2972 2979 2985 2992 2999 3006 3013
3020 3027 3034 3041 3048 3055 3062 3069 3076 3083
3090 3097 3105 3112 3119 3126 3138 3141 3148 3155
1
1
1
1
1
1 2 3 3
1 2 3 3
1 2 3 3
1 2 3 4
1 2 3 4
4
4
4
4
4
5
5
5
5
5
1 1 2 2 2 3 3 3
1 1 2 :I 2 3 3 4,
1 1 2 2 2 3 3 4,
1 1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
8 4
3
3
4
4
4
4
4
,4
1 1 2 2 3 3 4 4
1 2 2 3 3 4 4 Ii
1 2 2 3 3 4 4 5
1 2 2 3 3 4 4 Ii
1 2 2 3 3 4 4 Ii
1 2 2 3 3 4 5 5
"
2
2
2
2
2
2
2
2
3
3
4 5 II
4 5 8
4 IS 6
IS
5
5
6
6
6
6
6
6
6
456

ANTILOGARITHMs.-(Cantinued)
Proportional parts
Logarithms
"5
7'
11213141516171819
.50
.51
.52
.53
.54
3162 3170 3177 3184 3192 3199 3206 3214 3221 3228
3236 3243 3251 3258 3266 3273 3281 3289 3296 3304
3311 3319 3327 3334 3342 3350 3357 3365 3378 3381
3388 3396 3404 3412 3420 3428 3436 3443 3451 3459
3467 3475 3483 3491 3499 3508 3516 3524 3532 3540
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
3
3
3
3
3
4
4
4
4
4
4
5
5
5
5
5 6 7
5 6 7
5 6 7
6 6 7
6 6 7
.55
.56
.57
.58
.59
3548 3556 3565 3573 3581 3589 3597 3606 3614 3622
3631 3639 3648 3656 3664 3673 3681 3690 3698 3707
3715 3724 3733 3741 3750 3758 3767 3776 3784 3793
3802 3811 3819 3828 3837 3846 3855 3864 3873 3882
3890 3899 3908 3917 3926 3936 3945 3954 3963 3972
1
1
1
1
1
2
2
2
2
2
2
3
3
3
3
3
3
3
4
4
4
4
4
4
5
5
5
5
5
5
6
6
6
6
7
7
7
7
.60
.61
.62
.63
.64
3981 3990 3999 4009 4018 4027 4036 4046 4055 4064
4074 4083 4093 4102 4111 4121 4130 4140 4150 4159
4169 4178 4188 4198 4207 4217 4227 4236 4246 4256
4266 4276 4285 4295 4305 4315 4325 4335 4345 4355
4365 4375 4385 4395 4406 4416 4426 4436 4446 4457
1
1
1
1
1
2
2
2
2
2
3
3
3
3
3
4
4
4
4
4
5
5
5
5
5
6
6
6
6
6
6
7
7
7
7
7 8
8 9
.65
.66
.67
4467 4477 4487 4498 4508 4519 4529 4539 4550 4560
4571 4581 4592 4603 4613 4624 4634 4645 4656 4667
4677 4688 4699 4710 4721 4732 4742 4753 4764 4775
4786 4797 4808 4819 4831 4842 4853 4864 4875 488
4898 4909 4920 4932 4943 495/? 4966 4977 4989 5000
1
1
1
1
1
2
2
2
2
2
3
3
3
3
4
4
4
4
3 5
5
5
6
6
.68
.69
6
7
7
7
5012 5023 5035 5047 5058 5070 5082 5093 5105 5117 1 2 4 5 6 7
5129 5140 5152 5164 5176 5188 5200 5212 5224 5236 1 2 4 5 6 7
5248 5260 5272 5284 5297 5309 5321 5333 5346 5358 1 2 4 5 6 7
5370 5383 5395 5408 5420 5433 5445 5458 5470 5483
4 5 6 8
5495 5508 5521 5534 5546 5559 5572 5585 5598 5610 ~ ~ 4 5 6 8
.75
.76
.77
.78
.79
5623 5636 5649 5662 5675 5689 5702 5715 5728 5741
5154 5768 5781 5794 5808 5821 5834 5848 5861 5875
5888 5902 5916 5929 5943 5957 5970 5984 5998 6012
6026 6039 6053 6067 6081 6095 6109 6124 6138 6152
6166 6180 6194 6209 6223 6237 6252 6266 6281 6295
6310 6324 6"389 6353 6368 6383 6397 6412 6427 6442
6457 6471 6486 6501 6516 6531 6546 6561 6577 6592
6607 6622 6637 6653 6668 6683 6699 6714 6730 6745
6761 6776 6792 6808 6823 6839 6855 6871 6887 6902
6918 6934 6950 6966 6982 6998 7015 7031 7p47 7063
.80
.81
.82
.83
.84
.85
.86
.87
.88
.89
8
8
8
8
I)
8 9
8 9
5 6 7 8 9
.70
.11
.72
.73
.74
6 7 7
7
8
8
8
910
910
910
910
8 9
810
910
910
910
11
11
11
11
12
1
1
1
1
1
3 4 5 7 8 9101 2
3 4 5 7 8 9 1112
3
7 810 1112
3
7 810 1113
3 4 6 7 910 1113
1
2
2
2
2
3
3
3
3
! Ig
3 4 6 7 910 1218
5
8 911 1214
5 ~ 8 9 11121 4
5 6 8 9 111314
5 6 810 11131 5
7079 7096 7112 7129 7145 7161 7178 7194 7211 7228 2 3 5 7 810 12131 5
7244 7261 7278 7295 7311 7328 7345 7362 7379 7396 2 3 5 7 810 1213 15
.90
.91
.92
.93
.94
7413 7430 7447 7464 7482 7499 7516 7534 7551 70682
7586 7603 7621 7638 7656 7674 7691 7709 7727 7745 2
7762 7780 7798 7816 7834 7852 7870 7889 7907 7925 2
7943 7962 7980 7998 8017 8035 8054 8072 8091 8110 2
8128 8147 8166 8185 8204 8222 8241 8260 8279 8299 2
8318 8337 8356 8375 8395 8414 8433 8453 8472 8492 2
8511 8531 8551 8570 8590 8610 8630 8650 8670 8690 2
8710 8730 8750 8770 8790 8810 8831 8851 8872 8892 2
.95
.96
.97
.98
.99
8913 8933 8954 8974 8995 9016 9036 9057 9078 9099
9120 9141 9162 9183 9204 9226 9247 9268 9290 9311
9333 9354 9376 9397 9419 9441 9~62 9484 9506 9528
9550 9572 9594 9616 9638 9661 9683 9705 9727 9750
9772 9795 9817 9840 9863 9886 9908 9931 9954 9977
2
2
2
2
2
3 5 7 910 1214 16
4 5 7 911 1214 16
4 5 7 911 13 14 16
4
4
4
4
4
6
6
6
6
6
7 911 13 15 17
8 911 13 15 17
8 10 12 14 15 17
810 12 14 16 18
8 10 12 14 16 18
4 6 810 12 15 1719
4 6 811 13 15 1719
4 7 911 13 15 1720
4 7 911 13 16 1820
5 7 911 14 16 18 2()
INDEX
A
~ldehyde
content, assay of volatile

oils for, 372
Abbe refractometer, 242
Alkalimetry, 86
direct titration methods, 86
diagram of, 243
table of official substances asillustration of, 242
sayed by, 91
temperature control for, 244
residual titration methods, 90
Absolute viscosity, 257
table of substances assayed by,
Absorption pipette, 191
103
Acacia, moisture content, deterAlkaloidal assaying, 386
mination of, 331
by aliquot-part method, 403
Accuracy and honesty, 3
decanting the aliquot portion,
Acetylization flask, 368
Acetylsalicylic acid tablets, assay of,
396
determination of the alkaloidal
111
content, 401
Acid number, 245
extraction of the drug, 395
Acid value, 345
shaking out with acid, 398
definition of, 345
with immiscible solvent, 399
determination of, 347
table of official substances astable of official substances with
sayed by, 411
limits, 348
emulsification in, 389
Acid-base equilibrium, 272
general procedures, 394
Acidimetry, 105
type methods, 403
and alkalimetry, 65
principles applied in, 386
sources of error in, 390
by special methods, 418
112
by total-extraction method, 411
residual titration methods, 110
percolator recommended for,
398
114
table of official substances asAcid-insoluble ash, 323
sayed by, 416
Acidity index, 345
Alkaloidal salts, assay of, 431
Adsorbents in alkaloidal assays, 402
table of those official salts assayed
Alcohol, determination of, 214
for content of alkaloid, 436
Alcoholic potassium hydroxide, 365
Alkaloidal test solutions, 393
Alcohol-soluble extractive, 339
Alkaloidal titrations, choice of inditable of official substances with
cators for, 392
limits of, 340
Alkaloids, gravimetric determinaAlcohols, assay of volatile oils for,
tion of, 403
368
volumetric determination of, 407
457
458
Allyl isothiocyanat'e, assay for, 382

Aloe, assay of, 341
Alum, assay of, 50
Aluminum, determinatioIl of, 50
Ammonia in water, 261
Ammonium acetate solution, assay
of,97
Ammonium bromide, assity.of, 124
Ammonium hypophospbite, assay
of, 183
Ammonium thiocyanate 0.1 N solution of, 118
Ampere, 305
Amylopsin, 448
Analytical balance, 21
diagram of, 22
figure of, 22
rules for use and care of, 23
sensitivity of, 24
zero point determination, 24
Analyzer, 248
Angle of incidence, 240
of refraction, 240
of rotation, 248
Anions, 307
Anode, 307
illustrations of, 309
rotating, 311
Antilogarithms, 455
Areca, assay of, 409
Arithmetical mean, 10
average deviation hom, 10
Aromatic sulphuric acid, assay of,
110
Arsenic trioxide, assay of, 162
nephelometric estimation of, 267
Ascaridol content, assay for, 380
Ash content, 323
Ash limits of official substances, 327
Atomic weights, table of, inside back
cover
Avogadro's hypothesis, 189
B
Babcock bottle, 384
Balance, analytical, 21
Barium hydroxide, 83
standard solution of, 83
Barium sulfate, determination of
specific gravity of, 220
Belladonna leaf, assay of, 416
extract, assay of, 415
fluidextract, assay of, 416
tincture, assay of, 414
Benzoin, assay of, 339
Block comparator, 299
Boiling points, 226
apparatus for, 234
method of determining, 235
table of official, 238
Boric acid, asslJlY of, 107
determination of pH of solution
of,287
of solubility in water, 205
Boyle's law, 189
Bromcresol purple, 73
Bromine, 0.1 N, 180
Bromphenol blue, 73
Bromthymol blue, 73
Buffer mixtures,,294
table of, 294
Buffer solutions, 293
solutions used to prepare, 293
Buffers, 67
'
Bunsen valve, 150
Burettes, 58
calibration of, 63
official requirements ior, 59
reading of, 60
C
Caffeine, assay of drugs containing,
429
table of official substances assayed
for, 431
Calcium carbonate, assay of, 146
Calcium gluconate, assay of, 141
Calcium glycerophosphate, assay of,
48
Calculation of results and errors, 9
Calibration of weights, 26
IlY DEX
459
Calomel electrode, 279

Copper sulphate, assay of, 172
illustration of, 279
Cottonseed oil, saponification value,
potential of, 280
determination. of, 349
solidification temperature of fatty
Camphor, determination of specific
gravity of, 221
acids. of, 231
Cantharides, assay of, 437
Coulomb, 306
Critical angle, 241
Carbon dioxide, assay of, 192
Critical ray, 241
Cathode, 307
Crocus, assay for color, 265
Cations, 307
Crucibles, 16
Centipoise, 257
diagram of position above flame,
Charles' law, 190
20
Chloride ion, determination of, 37
fritted glass, 17
Chlorides, turbidimetric test for,
Gooch filtration, 16
270
Crude fiber, 336, 342
Chlorinated lime, assay of, 170
Cubic centimeter, 58
Cinchona, assay of, 405
Cupric sulphate, assay of, 172
compound tincture, assay of, 406
Citrated caffeine, 431
D
Cleaning mixture, 5
Clove, crude fiber content, deterDalton's Jaw, 198
mination of, 342
Decomposition voltage, 308
ether-soluble extractive content,
Density, 207
Desiccators, 21
Cochineal, 74
Deviation, average, 10
Coefficient of distribution, q91
Dextrorotatory, 248
Colchicum, assay of, 422
Dichromate methods, 151
table of official substances assayed
Digitalis leaf, ash content, determiby the same method as, 425
nation of, 225
Color, assay for, 265
moisture content, determination
Color comparators, 298
of,331
Color standards, 297
.Diluted sulfuric acid, assay of, 106
Colorimeters, 263
Diphenylamine indicator, 152
Colorimetric methods, 260
Direct titration, 86
Colorimetry, 260
Dis1J.lling points, 234
Common ion effect, 35
apparatus for, 234
Comparators, color, 298
Compound solution of iodine, assay
of volatile oils, 363
of, 164
Distribution constant, 391
Congealing points, 226
'v Distribution ratio, 391
Duboscq colorimeter, 262, 263
E
Congo red, 74
Constant weight, 20
Economy of time, 5
Constants of fats, oils, and waxes,
Electrical units, 305
345
Electroanalysis, 307
460
Electrochemical equivalent, 306

Electrode,'glass, 290
hydrogen, 277
potential, 280
quinhydrone, 288
Electrolysis, 307
Electrolyte, 306
Electrolytic apparatus, 309
diagram of connections for, 311
Electrolytic methods, 305
assay of copper sulphate, 313
assay of mercuric chloride, 315
319
Electron, 130
Elixir of iron, qumme and strychnine, pH determination of, 289
Elixir of three bromides, assay of,
125
End point, definition of, 57
in dichromate methods, 152
in precipitation methods, 116
Engler degrees, 259
Enzymes, assay of substances containing, 444
factors influenCing activity, 444
Ephedrine sulfate, assay of, 435
Epinephrine hydrochloride, estimation of colorimetric ally, 264
Equivalence point, 65
Errors, indeterminate, 10
nature of, 9
Ester content, assay of volatile oils
for, 364
Ester number, 353
definition of, 353
official substances with required,
353
Ester value, 353
Ether-soluble extractive, 337
table of official substances with
limits of, 338
Ethyl nitrite, assay of spirit of, 178
Eucaine hydrochloride, assay of, 432
Evaporation of liquids, 19
Extract of beef, estimation of

nitrogen in, 100
Extraction, with immiscible solvents, 399
Extractive, alcohol-soluble, 339
ether soluble, 337
purified petroleum benzin, 341
water-soluble, 341
Extractive content, 336
F
Faraday's laws, 130,306

Fats, constants of, 345
Ferric ammonium sulfate, indicator
solution, 116
Ferric chloride, assay of, 168
Ferrous carbonate, assay of mass of,
153
Ferrous sulfate, assay of, 136
Fiber, crude, 237
Figures, significant, 11
Flasks, calibratio~_of, 63
Fluidextract, of belladonna leaves,
determination of alcohol content of, 214
I
of hyoscyamus, assay of, 415
Fractional distillation of volatile oils,
363
Fritted glass crucibles, 17
G
Gas burette, 191
Gas pipette, 191
Gases, laws of, 189
Gasometric methods, 188
193
General operations, 16
Glass crucibles, fritted, 17
Glass electrode, 290
Graduated cylinders, 62
Gravimetric analysis, 29
theory of, 29
Gravimetric determinations of alkaloids,403
461
INDEX
Gravimetric methods, 37
assay of alum, 50
of calcium glycerophosphate, 48
of mercuric chloride, 45
of sodium chloride, 37
of sodium sulfate, 43
of solution of magnesium
citrate, 52
sample data for, 40
Grazing incidence, 241
Guarana, 429
H
Hydrastis, assay of, 403

Hydriodic acid, assay of, 124
Hydrochloric acid, assay of, 105
normal solution of, 79
potentiometric titration of, 281
Hydrocyanic acid content, assay of
volatile oils for, 378
Hydrogen electrode, 277
platinization of, 278
Hydrogen ion concentration, 66, 272
of some official substances, 301
Hydrogen ion measurements, colorimetric methods of, 292
potentiometric methods of, 277
precautions during, 285
scheme of assembly for, 280
Hydrogen peroxide, assay 01 solution
of, 139
Hyoscyamus, assay of, 411
extract of, assay of, 415
fluidextract of, assay of, 415
tincture of, assay, 414
Indicator solutions, sensitivity of, 76

Indicators, 70, 296
for alkaloidal titrations, 392
definition of, 70
diphenylamine, 152
ferric ammonium sulfate, 116
mixed,296
potassium chromate, 117
lor precipitation methods, 116
rules for use of, 73
starch solution, 157
table of commonly used, 71
theory'of,70
universal, 296
used in colorimetric hydrogen ion
measurements, 296
Iodine, assay of compound solution
of, 164
number, definition of, 355
official substances with requirements for, 359
0.1 N solution of, 159, 160
standardization against 0.1 N
sodium thiosulfate, 160
test solution, 394
titration flask, 358
value, 355
definition of, 355
determination, 357
Iodobromide test solution, 356
Iodometric methods, 155
Ionization, theory of, 29, 69
Ipecac, assay oi, 400
J
Jalap, assay of, 440
K
I
Immiscible solvents, 388

extraction with, 399
use in non-alkaloidal assays, 437
Indeterminate errors, 10
Index of refraction, 240
Indicator solutions, 73
Ketone content, assay of volatile

oils for, 375
Kinematic viscosity, 258
Kjeldahl apparatus, 99
Kettsdorfer number, 349
Koppescharr's solution, 180
462

L
Laevorotatory, 248
Laws, Boyle's, 189
Charles', 190
Dalton's, 198
Faraday's, 98, 306
Ohm's, 305
Limit test for chlorides and sulfates,
269, 270
Liquid petrolatum, determination of
kinematic viscosity of, 259
Liquids, transfer of, 19
Liter, normal, 58
Litmus, 74
Logarithms of numbers, 453
use of, 13
Lunge nitrometer, 194
Nature of errors, 9
Nephelometry, 260, 266
Nessler apparatus, 260
Neutralization methods, 65
buffers in, 67
pH value in, 66
theory of, 66
Neutralization reactions, 69
Nitrogen estimation, by GunningKjeldahl method, 98
with nitrates and nitrites absent,
98
with nitrates present, 99
Nitrometer, illustration of, 194
preparation and testing of, 195
Non-volatile ether-soluble extractive, 337
Normal conditions, 189
Normal liter, 42
Normal solution, 7(,-77
Normality, 77
Normality factor, 78
Notebooks,4
,.
Nux vomica, assay of, 425
M
Magnesia magma, assay of, 94
Magnesium, determination of, 52
Magnesium citrate, assay of, 52
Mass of ferrous carbonate, assay of,
153
Materials required, 7, 392
Mayer's reagent, 394
Measuring flasks, 61
Melting points, 226
apparatus for taking, 226
Meniscus, 60
>to
Mercuric chloride, assay of, 45
~.!"
Mercuric potassium iodide, 394
Mercurous chloride, assay of, 166
Methenamine, assay of, 95
Methyl orange, 74
Methyl red, 75
Milliliter, 58
Moisture content, 329
Moisture limits of official substances,
333
Moisture tube, 332
Molal solution, 76
Molar solution, 76
preparation of, 185
Monochromatic light, 249
o
Official sample, 8
Ohm, 305
Oil of bitter almonds, assay for
benzaldehyde content, 373
assay for hydrocyanic acid, 379
of caraway, assay of, 375
of chenopodium, assay of, 380
of clove, assay of, 377
of mustard, assay of, 382
of orange, determination of refractive index of, 245
of peppermint, assay for total
esters, 366
assay for total menthol, 368
nephelometric estimation of,
269
Oleoresin of aspidium, assay of, 439
INDEX
Olive oil, iodine value determination
of,357
Opium, assay of, 418
table of official substances'assayed
by the same method as, 422
Optical activity, official substances
with requirements, 254
Optimum temperature of enzymes,_
444
Organic solvents, evaporation of, 400
Oxalic acid, 0.1 N solution of, 143
Oxidation, definition of, 130
Oxida tion-red uction, dichromate
methods, 151
154
140
iodometric methods, 155
standard solutions for, 132
methods, 155
theory of, 129
Oxide, official substances assayed by
ignition to, 51
P
Pancreatin, assay for casein digestive power, 449
for starch digestive power, 447
Parallax, effect of, 60
Partition coefficient, 391
Pepsin, assay of, 445
Permanganate methods, 133
pH, colorimetric methods of determining, 292
indicators and their use, 293
measurements, directions for, 29.6
meter, 291
potentiometric methods of determining, 277
relationship to voltage, 283, 284
value, 66
Phenobarbital, assay of tablets of,
441
Phenol, assay of, 181
463
Phenol content, assay of volatile

oils for, 377
Phenol red, 75
Phenolphthalein, 75
Photometric methods of analysis,
260
Physiological methods, 321
Physiological salt solution, determination of pH of solution of, 287
Pipettes, 60
calibration of, 63
illustrations of, 62,
official requirements for, 60
Plane-polarized light, 247
Poise, 257
Polarimeter, 250
diagram of optical parts of, 250
of Laurent, 251
of Schmidt and Hansch, 251
Polariscope, 251
Polariscope tube, 248
Polarization, definition of, 247
Polarizer, 248
Policemen, 6
Potassium and sodium tartrate,
assay of, 92
Potassium chloratEJ, assay of, 149
Potassium chromate, indicator solution,117
Potassium dichromate, 0.1 N solution of, 152
Potassium hydroxide, alcoholic,
preparation of, 365
Potassium iodate, 185
molar solution of, 185
official substances assayed with,
"
187
Potassium iodIde, assay of, 186
test solution, 356
Potassium mercuric iodide test solution, 394
Potassium permanganate, 0.1 N
solution of, 134
Potential, electrode, 280
Potentiometric methods, 277
464
Potentiometric titration, of acetic

acid with sodium hydroxide, 287
of hydrochloric acid sodium with
hydroxide, 281
Precipitates, 17
colloidal apd fine-grained, 18
drying and ignition of, 19
filtration and washing of, 17
Precipitation methods, 116
by direct titration, 119
121
end point in, 116
indicators for, 117
by residual titration, 122
127
Probability curve, 10
Proteolytic power, assay of pepsin
for, 445
Proton, 130
Proximate assays, 386
Purified petroleum benzin extractive,
341
Purity and strength requirements, 6
Pycnometer, 208
Geissler type, 210
method of filling, 209
Sprengel-Ostwald, 208
Pyrophosphate, table of official substances assayed as, 54
Q
Quinhydrone, 288
Quinhydrone electrode, relation of
pH to voltage, 288
R
Reactions, in neutralization, 69
reversible, 30
Reagents, 6
Reduced iron, assay of, 137
Reduction, definition of, 130
Refractive index, 240
Refractive indices, determination of,
245
Refractive indices, table of official

substances with, 246
Refractometers, 241
Relative viscosity, 257
Rennin, assay of, 450
Residual titration, 57, 86
Residue, official requirements of, 326
Result, rejection of, 11
Results and errors, 9
Reversible reactions:' 30
Riders, 28
Rosin, acid value, determination of,
347
Rotating anode, device for, 311
Rotatory power, 247
of vola tile oils,. 362
Roulette comparator, 300
S
Salicylic acid, melting point determination o,"227
Samples, 'selection of, 7
drying and ignition'of,\19
Sampling, 7
Saponification value, definition of,
349
official substances with limits of,
352
Saybold universal viscosity, 259
Sedimentation cone, 447
Separatory funnel, 388
Significant figures, 11
Silver nitrate, 0.1 N solution of, 117
Sodium arsenate, exsiccated, assay
of, 173
Sodium bicarbonate, assay of, 86
Sodium chloride, assay of, 37, 122
Sodium hydroxide, assay of, 87
normal solution of, 81
Sodium nitrite, assay of, 144
Sodium salicylate, assay of, 89
tablets, assay of, 108
Sodium sulfate, assay of, 43
INDEX
465
Specific gravity determinations, by

Sodium thiosulfate, 0.1 N solution,
means of pycnometer, 214
157
of
liquids by means of the WestSolidification temperature, deter~
phal balance, 216
mination 6, 231
by the use of hydrometers, 218
table of, 233
by weighing a solid of known
Solubility, 203
specific gravity in, 218
apparatus used to determine, 204
rotation, definition of, 249
Specific
descriptive terms used in, 203
Specific rotatory power, calculntion
factors affecting, 204
of,249
product, pril!;ciple, 31
table of, 32
of volatile ,?illl, 364
Solution of arsenouS acid, nephel- Spirit of ethyl nitrite, assay of, 179,
196
'
ometric estimation of arsenic
of peppermint, assay of, 384
trioxide in, 267
nephelometric estiml),tion of oil
of epinephrine hydrochloride, de~
in, 269
termination of pH of, 264
Standard conditions, 189
Solutions, classes of, 203
Standard solutions, 57, 76, 132, 157
concentrations ~f standard, 76
of hydrochloric acid, 79
of indicators, 73
in oxidation-reduction methods,
molal,76
132
molar, 76
of sodium hydroxide, 81
normal,76
of sulfuric acid, 83
Source and nature of errors, 9
0.1 N ammonium thiocynnate, 118
Soxhlet extraction apparatus, 337
0.1 N barium hydroxfde, 83
Special methods, alkaloidal assays
0.1 N bromine, 180
by, 418
0.1 N iodine, 159, 160
Specific gravity, 207
0.1 N oxalic acid, 143
of fats and oils, 211
0.1 N potassium dichromate, 152
'official method for, 211
0.1 N pot!tssiunl permanganate.
of liquids, 208
134
methods of determining, 208
0.1
N
silver nitr!1te, 117
method of determining alcohol,
0.1 N sodium thiosulfate, 157
211
Standardiziltion, 76
of solids, 219
Starch digestive power, assllY of
heavier than and insoluble in
pancreatin for, 447
water, 219
indicator solution, 157
heavier than and soluble in
test solution, 356
water, 222
in ~ Stoichiometric point, 58, 65
lighter than and insoluble
Strong silver-protein, assay of, 120
water, 220
Success as an analyst, 3
lighter than and soluble in Sucrose, determination of specific
rotation of, 252
water, 223
Sulfate, determination of, 42
official substances determined as,
44
SpeCific gravity bottle, 208
466
Sulfide, determination of, 45

official substances determined as,
47
Sulfuric acid, preparation of a 10 per
cent solution of, 219
standard solution of, 83
T
Tar~aric acid, assay of, 110
Test solutions, for alkaloids, 394
Theobromine with sodium salicylate,
assay of, 432
Xheophylline with sodium acetate,
assay of, 434
Thymol blue, 75
Thyroid, assay of, 170
Titer, 65
Titration, 78
curves, 285
direct, 86
potentiometrically, 281
residual, 86
Titrimetric methods, 57
Toluene moisture method, 331
Total ash, 323
Total esters, assay of oil of peppermint for, 364
Transfer of liquids, 19
Turbidimetric tests, 269
Type process, aliquot, 403
alkaloidal assays by, 403
general procedure, 394
Type process, total extraction, for
alkaloidal assays, 403
Units of capacity, 58
of electricity, 305
Unsaponifiable matter, 354
V
ViSCOSimeter, 258
ViSCOSity, definition of, 256
units of, 257
Volatile ether-soluble extractive, 337
Volatile oils, assay of, 361

for alcohol content, 368
for aldehyde content, 372
for ester content, 364
for hydrocyanic acid content,
378
for ketone content, 375
for phenol content, 377
in spirits, 384
congealing points of, 363
constituents of, 361
determination of specific gravity
of,217
distilling points of, 363
fractional distillation of, 363
refractive index of, 363
rotatory power of, 362
solubility of, 364
specific gravity ~f, 362
Volt, 305
Voltage, relationship, to pH, 284
Volumetric analysis, 57
Volumetric apparat~!l, 58
calibration of, 62
cleaning of, 62
sources of error in use of, \64.
W
Wagner's reagent, 394
Wash bottles, 5
Water, determination of aIIJ.monia
content of, 261
Water-soluble extractive, 341
table of official substances with
limits of, 341
Wax, yellow, determination of specific gravity of, 222
Weights, calibration of set of, 26
description of, 25
Westphal balance, 216
use of, 217
X
Xylene moisture method, 331
z
Zinc oxide, assay of, 92

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QUANTITATIVE

GLENN L. JENKINS, PH.D.

ANDREW G. DuMEZ, PH.D.

McGRAW-HILL BOOK COMPANY,

. .f.',x._: .0; ~I,::

~.. ~:oom No.

Rec~lved~:t-'5""1eq~~ ~ :g'6ok No,

McGRAW-HILL PUBLICATIONS IN PHARMACY

SERIES OF TEXTS AND REFFERENCE WORKS

Professor of Pharmacy, University of North Carolina,

Associate Professor of Pharmaoy, State University of

D'l"n, Ohio State University, College of Pharmaoy.

Dean, University of Oklahoma, Sohool of Pharmacy.

Dean, University of Florida, School of Pharmacy.

Dean, Medical College of Virginia. Sohool of Pharmaoy.

Associate Professor of Pharmaoy, University of Michigan, College of Pharmaoy.

Profeasor of Biology and Pharmacognosy, Philadelphia

MCGRAW-HILL BOOK COMPANY, INC.

All rights reserved. This book, or

TEXT OF THE UNITED STATES PHARMACOPOEIA IS ElY VIRlUE

1"HE BOARD OF TRUSTEE;s

OF THE UNITED STATES PHARMACOPOEIAL. CONVENTION.

RESPONSIBLE FOR ANy

FOR ANY ERRORS IN THE

PERMISSION TO USE FOR COMMENT PARTS OF' THE TEXT OF'

THE NATIONAL FORMULARY, SIXTH EDITION, IN THIS VOLUME

THE AUTHORITY OF THE COUNCIL OF 'l'HE AMERICAN PHARM4._

THE MAPLE PRESS COMPANY, YORK, PA.

PREFACE TO THE SECOND EDITION

PREFACE TO THE SECOND EDITION

PREFACE TO THE FIRST EDITION

PREFACE TO THE FIRST EDITION

in addition to the exercises pertaining to the analytical. balance

PREFACE TO THE SECOND EDITION.

PREFACE TO THE FIRST EDITION

Definitions and scope of quantitative pharmaceutical chemistryReferences.

Sampling-Calculation of results and errors-General operationsThe analytical balance-Weights.

Theory of ionization-Reversible reactions-Solubility product

Assay of sodium chloride, of sodium sulfate, of mercuric chloride, of

Definitions-Volumetric apparatus-The calibration of volumetric

Direct titration methods: Assay of sodium bicarbonate, of sodium

Residual titration methQds: Assay of zinc oxide, of potassium and

Direct titration methods: Assay of diluted sulfuric acid, of boric

Determination of the end point-Indicators. Standard solktions.

Starch indicator solutions. Standard solutions: Preparation and

Titration of the iodine liberated from potassium iodide with sodium

Theory...--Apparatus-Test of the nitrometer-Assay of carbon

Methods used to determine the specific gravity of liquids: The use

Melting point: Determination of the melting point of salicylic acid.

Refractive index: Refractometers-The Abbe refractometer.

Definitions-Apparatus: The Saybolt viscosimeter.

Acid base equilibrium and pH.

in Official Pharmaceutical Analyses

Assay for ketone content: Assay of oil of caraway.

General principles: Sources of error-Theory of distribution

Alkaloidal assays by aliquot-part method.

TABLE OF ATOMIC WEIGHTS . . . . . . . . . . . . lnside back cover

gases when a suitable reagent is used to remove one of the

6. FALES, "Inorganic Quantitative Analysis," D. Appleton-Century

6. WILKINSON, "Calculations in Quantitative Analysj.s," McGraw-Hill

Company, Philadelphia, 1926.

"Plant Analysis, Qualitative and Quantitative,"

1. BLYTH, "Foods, Their Composition and Analysis" 6th ed., D. Van