Classification Tests for Hydroxyl and Carbonyl Compounds

University of Santo Tomas

Faculty of Pharmacy
2FPH Group 5
Members: Mondoedo, Samantha; Lopez, Princess; Maternal, Charles; Miranda, Joyce; Nasinopa,
Reina; Navata, Jemina

ABSTRACT
The aim of the experiment was to be able to distinguish whether a compound is hydroxyl
or carbonyl containing,differentiate the three types of alcohols, differentiate aldehydes from
ketones and to explain the mechanisms involved in the differentiating tests. Therefore in this
experiment the proponents were able to classify the compounds based on the different tests. The
samples were analyzed through tests involving the solubility of alcohols in water, Lucas test,
Chromic Acid test, 2,4-Dinitrophenylhydrazone (2,4-DNP) test, Fehlings test, Tollens Silver
Mirror test, and Iodoform test. Lucas test differentiates tert-butyl alcohol (primary), sec-butyl
alcohol (secondary) and n-butyl alcohol (tertiary) . Chromic test gave a positive result for nbutyl alcohol, acetaldehyde, acetone, isopropyl alcohol. 2,4-Dinitrophenylhydrazone test was
performed for aldehydes and ketones; all samples gave a positive result. Fehlings Test and
Tollens Silver Mirror Test are tests for aldehydes. These tests prove that acetaldehyde and
benzaldehyde are aldehydes while acetone and acetophenone are ketones. Iodoform test is a test
for methyl carbinol and methylcarbonyl groups.

In organic chemistry, functional groups are

specific groups of atoms within molecules that
are responsible for the characteristic chemical
reactions of those molecules and examples of
functional groups are hydroxyl and carbonyl
groups.

classified as primary (1), secondary (2), or

tertiary (3), depending on the number of carbon
substituents bonded to the hydroxyl-bearing
carbon [4]. Hydroxyl groups are especially
important in biological chemistry because of
their tendency to form hydrogen bonds both as
donor and acceptor. This property is also related
to their ability to increase hydrophilicity and
water solubility[1].

An alcohol is a compound that has a

hydroxyl group bonded to a saturated, sphybridized carbon atom, R-OH. Alcohols are

Carbonyl group is a family of functional groups

composed of a carbon atom double-bonded to an
oxygen atom: C=O. The group is a constituent

Introduction

of carboxylic acids, esters, anhydrides,

acylhalides, amides, and quinones, and it is the
characteristic functional group of aldehydes and
ketones [2].
The functional group of an aldehyde is a
carbonyl group bonded to a hydrogen atom
(Figure 1). The functional group of a ketone is a
carbonyl group bonded to two carbon atoms
(Figure 2). Because of the polarity of the
carbonyl group, aldehydes and ketones are polar
compounds and interact in the liquid state by
dipole-dipole interactions. As a result, aldehydes
and ketones have higher boiling points than
those of nonpolar compounds with comparable
molecular weight [3].

Figure 1. Structure of Aldehyde

Figure 3. General Mechanism of Lucas test

The Chromic acid test or also known as
Jones oxidation distinguishes primary and
secondary alcohols from tertiary. Chromic acid
will oxidize a primary alcohol first to an
aldehyde and then to a carboxylic acid and it
will oxidize a secondary alcohol to a ketone.
Tertiary alcohols do not react. The OH-bearing
carbon must have a hydrogen atom attached.
Since the carbon atom is being oxidized in
primary and secondary, the orange chromium
Cr6+ ion is being reduced to the blue-green Cr3+
ion[4].

Figure 2. Structure of Ketone

The Lucas reagent is an aqueous solution
of strong acid (HCl) and zinc chloride (ZnCl).
The alcohol starting material must be
sufficiently soluble in aqueous environments for
the reaction to take place. The reaction that
occurs in the Lucas test is an SN1 nucleophilic
substitution[5]. The acid catalyst activates the
OH group of the alcohol by protonating the
oxygen atom. The C-OH+ bond breaks to
generate the carbocation, which in turn reacts
with the chloride ion (nucleophile) to generate
an alkyl halide product. Figure 3 shows the
general mechanism for this SN1 reaction [4].

Figure 4. General Mechanism of Chromic Acid

Test
The 2,4-Dinitrophenylhydrazone test
serves as a derivative formation. Both aldehydes
and
ketones
react
with
2,4dinitrophenylhydrazine to form a solid 2,4dinitrophenylhydrazone (DNP) derivative. The
color of this derivative can also provide useful
structural information. If the solid is yellow, this
most often means that the carbonyl group in the
unknown is unconjugated. A reddish-orange
color most likely means that the carbonyl group
is conjugated. In a few cases, compounds in

which the carbonyl group is not conjugated

produce orange precipitates. The double bond
must be separated from the carbonyl by one
single bond only. If the double bond is further
away, it is isolated from the carbonyl and not
conjugated with the carbonyl[4].

the test is carried out in a clean glass test tube,

forms a mirror on the test tube. Ketones are not
oxidized by the Tollens reagent, so the
treatment of a ketone with Tollens reagent in a
glass test tube does not result in a silver mirror.

Figure 7. General Mechanism of Tollens test

Figure 5. General Mechanism of 2,4-DNPH

test
In Fehlings test, the presence of
aldehydes and not ketones is detected by
reduction of the deep blue solution of copper (II)
to muddy green solution, and then form a brickred precipitate of insoluble cuprous oxide
(Cu2O). This test is commonly used for reducing
sugars but is known to be not specific for
aldehydes[3].

The Iodoform test indicates the presence

of an aldehyde or ketone in which one of the
groups directly attached to the carbonyl carbon
is a methyl group. Such a ketone is called a
methyl ketone. In the Iodoform test, the sample
is allowed to react with a mixture of iodine and
base. Hydrogens alpha to a carbonyl group are
acidic and will react with base to form the anion,
which then reacts with iodine in this way to form
the triiodo compound, which the reacts with
more base to form the carboxylic acid salt plus
iodoform, a yellow precipitate. Formation of a
yellow precipitate therefore indicates the
presence of a methyl group directly attached to
the carbonyl. The mechanism of the iodoform
reaction is that of alpha-halogenation of a
carbonyl compound under basic conditions,
followed by nucleophilic displacement of the
resulting triiodomethyl group by hydroxide.

Figure 6. General Mechanism of Fehlings test

Tollens test, also known as silvermirror test, is a qualitative laboratory test used to
distinguish between and aldehyde and a ketone.
It exploits the fact that aldehydes are readily
oxidized, whereas ketones are not. Tollens test
uses a reagent known as Tollens reagent, which
is a colorless, basic, aqueous solution containing
silver ions coordinated to ammonia [Ag(NH3)2+].
Tollens reagent oxidizes an aldehyde into the
corresponding carboxylic acid. The reaction is
accompanied by the reduction of silver ions in
Tollens reagent into metallic silver, which, if

Figure 8. General Mechanism of Iodoform test

Objectives of this experiment:

To distinguish whether a compound is

hydroxyl or carbonyl containing

tubes, 2 drops of each of the samples

were added separately. The mixtures
were shaken and allowed to stand for
10 minutes. When no reaction
occurred, the test tubes were placed
in a beaker with warm water for 5
minutes. Observations were recorded.

To differentiate the three types of

alcohols
To differentiate aldehydes from ketones
To explain the mechanisms involved in
the differentiating tests

Experimental
A.

Materials
Lucas reagent,
chromic acid reagent,
95% ethanol,
Fehlings A and B,
Tollens reagent,
5% NaOCl solution,
iodoform test reagent,
2,4-dinitrophenylhydrazine,
Pasteur pipette,
test tubes,
beaker,
sample compounds (ethanol, n-butyl
alcohol, sec-butyl alcohol, tert-butyl
alcohol,
benzyl
alcohol,
nbutyraldehyde, benzaldehyde, acetone,
acetophenone, isopropyl alcohol and
acetaldehyde)

B. Methods
1.
Solubility of Alcohols in water
In the five test tubes, 10 drops
each of ethanol, n-butyl alcohol, secbutyl alcohol, tert-butyl alcohol and
benzyl alcohol were placed. 1 mL of
water was added dropwise in each of
the test tubes containing alcohol
while shaking the mixture after each
addition. When cloudiness resulted,
0.25 mL of water was added at a
time, with vigorous shaking, until a
homogenous dispersion appeared.
The total volume of water added was
noted. When no cloudiness resulted
after the addition of 2.0 mL water,
alcohol was noted down to be soluble
in water.
2.
Lucas Test
Five test tubes with 1 mL each of
freshly prepared Tollens reagent
were prepared. In each of the test

Three test tubes were prepared

and 1 mL of Lucas reagent was
dropped in each of the test tubes. On
the first test tube, 2-3 drops of nbutyl alcohol was added. The test
tube was shook vigorously for a few
seconds and the mixture was allowed
to stand at room temperature. The
two other test tubes were placed with
2-3 drops of sec-butyl alcohol and
tert-butyl
alcohol,
respectively,
shook vigorously and the mixtures
were allowed to stand. The rate of
formation of cloudy suspension or
the formation of two layers was
observed.
3. Chromic Acid test
Using the samples n-butyl
alcohol,
acetaldehyde,
acetone,
isopropyl alchohol one drop of each
liquid or small amount of the solid
sample was dissolved in 1 mL of
acetone in different test tubes. 2 drops
of 10% aqueous K2Cr2O7 solution and
5 drops of 6 M H2SO4 were added in
each of the test tubes.
4.

2,4-Dinitrophenylhydrazone
DNP/2,4-DNPH) test

(2,4-

One drop of a liquid sample

(acetone, acetaldehyde, benzaldehyde
and acetophenone) was placed in a
test tube and 5 drops of 95% ethanol
were added. After shaking well, 3
drops of 2,4-dinitrophenylhydrazine
were added. The solution was allowed
to stand for at least 15 minutes when
no yellow or orange-red precipitate
forms.

5. Fehlings test
Into each test tube, 1 mL of
freshly prepared Fehlings reagent
(made by mixing equal amounts of
Fehlings A and Fehlings B) was
placed. 3 drops of the sample to be
tested were added and the test tubes
were placed in a beaker of boiling
water. Changes were observed that
occurred within 10-15 minutes. The
test was performed on acetaldehyde,
acetone,
benzaldehyde
and
acetophenone.

ethanol
n-butyl
alcohol
secbutyl
alcohol
tertbutyl
alcohol
benzyl
alcohol

homogenous
dispersion
CH3CH2OH
0.25ml
Miscible
2.00 mL
Immiscible
1.25 mL

miscible

0.35 mL

miscible

2.00 mL

immiscible

6. Tollens Test
Five test tubes with 1 mL each of
freshly prepared Tollens reagent were
prepared. In each of the test tubes, 2
drops of each of the samples
(acetaldehyde, acetone) were added
separately. The mixtures were shaken
and allowed to stand for 10 minutes.
When no reaction occurred, the test
tubes were placed in a beaker with
warm water for 5 minutes.
Observations were recorded.
7. Iodoform test
2 drops of each sample (acetone,
benzaldehyde and isopropyl alcohol)
were placed in different test tubes. 20
drops of 10% KI solution were added.
While shaking, 20 drops of fresh
chlorine
bleach
(5%
sodium
hypochlorite) were added slowly to
each tube and mixed. The formation
of a yellow precipitate was noted.
Results and Discussion
A. Solubility of Alcohols in Water
Table 1. Solubility of Alcohol in Water
Alcohol

Condensed
structural
formula

Amount of
water (in
mL) needed
to produce a

Solubility
to water

As shown in the table, only benzyl

alcohol and n-butyl alcohol were insoluble in
water, while ethanol, sec-butyl alcohol and tertbutyl alcohol were all soluble in water. This
follows the principle like dissolves like. Thus,
it can be said that all of these soluble compounds
are polar since water is polar. Of the alcohols
that were soluble in water, their required amount
of water to be added in order to be considered
soluble varies. This indicates that there are other
factors affecting solubility. One of these is the
presence of number of carbon atoms. The lower
the number of carbon atoms present, the more
soluble or more miscible a substance is.
Branching of carbon chains also affect
solubility. The more branching present, the more
soluble a compound is. This is only true for
organic compounds that have the same number
of carbon atoms present[2].
B. Lucas test
Table 2. Lucas test
Sample
n-butyl alcohol
sec-butyl alcohol
tert-butyl alcohol

Reaction observed
Clear solution
slightly turbid solution
Turbid solution

According to the table above, n-butyl

alcohol was soluble in Lucas reagent while secbutyl alcohol and tert-butyl alcohol were
observed to have a formation of cloudy layer.
Tert-butyl alcohol took the shortest time to form
the layer while sec-butyl alcohol took the

longest time. The reaction mechanism involved

in the Lucas test is based on SN1 reaction.
According toMcMurry, depends on the
formation of stable carbocations. Reactivity of
alcohols in SN1 reaction is 3 > 2 > 1.

E. Fehlings Test

C. Chromic Acid

benzaldehyde

Table 5. Fehlings test

Sample
acetaldehyde

Table 3. Chromic Acid test

Sample
n-butyl alcohol
Acetaldehyde
Acetone
Isopropyl alcohol

Reaction observed
blue-green solution
blue-green solution
blue-green solution
blue-green solution

Base on the results, n-butyl alcohol,

acetaldehyde, acetone, isopropyl alcohol
gave a positive result of blue-green solution.
Chromic Acid test involved reductionoxidation or redox reaction. 1 and 2
alcohols and aldehydes underwent oxidation
and chromium underwent reduction from
Cr6+ to Cr3+. 1 and 2 alcohols and
aldehydes reduced the orange-red chromic
acid/sulphuric acid reagent to an opaque
green or blue suspension of Cr (III) salts.[3]
D. 2,4 DNP test
Table 4. Results of the 2,4 DNP test
Sample
Acetaldehyde
Benzaldehyde
Acetone
Acetaphenone

Reaction observed
Yellow orange ppt
Yellow orange ppt
Yellow orange ppt
Yellow orange ppt

Table 4 shows the reaction of the samples to

2,4-DNP test. All the samples exhibited a
positive result because they all formed a yellow
orange precipitate. Hence, 2,4-DNP test proved
that the samples are carbonyl-containing
compounds and are either aldehydes or ketones.

acetone
acetophenone

Reaction observed
Reddish brown
solution with brickred ppt
Formed two layers
yellow solution and
light blue solution
with brick-red ppt
blue solution with no
ppt
blue solution

As shown in the given table, only acetone

and acetophenone did not react to form a
precipitate while the rest gave a positive result
of brick-red precipitate. Fehlings test involved
reduction-oxidation
or
redox
reaction.
Aldehydes were oxidized to carboxylic acids
while ketones did not undergo oxidation. In
here, copper was reduced from Cu2+ to Cu1+[2].
F. Tollens Test
Table 6. Tollens test
Sample
acetaldehyde
acetone

Result observed
+
-

Table 6. Tollens Silver Mirror test

Sample
acetaldehyde
n-butyraldehyde
benzaldehyde
acetone
acetophenone

Result observed
silver mirror
flesh solution
light yellow solution
with globules
dark-gray solution
turbid gray solution

According to Table 6, only acetaldehyde

formed a silver mirror. The Tollens Silver
Mirror test involved reduction-oxidation or
redox reaction. Aldehydes were oxidized to

carboxylic acids while ketones did not undergo

oxidation except alpha-hydroxyketone. Silver
was reduced from Ag1+ to Ag0 [3].
G. Iodoform Test
Table 7. Iodoform test
Sample
benzaldehyde
acetone
isopropyl alcohol

Reaction observed
Forms an oily layer
Forms yellow ppt
Forms yellow ppt

Base on the given table, acetone and

isopropyl alcohol gave a result of yellow
precipitate. In this test, yellow crystals or
precipitate gave a positive result. An alkaline
solution of sodium hypoiodite, formed from
sodium hydroxide and iodine, converted
acetaldehyde and aliphatic methyl ketones into
iodoform (haloform reaction). Since the reagent
was also an oxidizing agent, alcohols which are
readily oxidized to acetaldehydes or methyl
ketones also gave a positive reaction. The
mechanism of iodoform synthesis occurred
through a series of enolate anions, which are
iodinated; hydroxide displaced the Cl3- anion
through an addition/elimination pathway[4].

References:
[1]
Hydroxyl
Group.
(2014).
In
Encyclopdia Britannica. Retrieved October
23, 2010, from Encyclopdia Britannica
Online:
http://www.daviddarling.info/encyclopedia
/P/phenol.html.
[2] Shriner, Fuson, Curtin. Systematic
Identification of Organic Compound: A
Laboratory Manual Fifth Edition. John
Wiley & Sons, Inc. New York: Van
Hoffmann Press
[3] Brown, W., Poon, T. (2014).
Introduction
to
Organic
Chemistry
International Student Version (7th edition).
NJ, USA: John Wiley & Sons, Inc.

[4] McMurry, J. (2010). Foundations of

Organic Chemistry (Philippine edition).
USA: Cengage Learning Asia Pte. Ltd.
[5] Zumdahl, S., Zumdahl, S. (2013).
Chemistry: An Atoms First Appraoch
(International edition). USA: Brooks/Cole,
Cengage Learning.
[6] Bathan,G., Bayquen,A., Cruz,C.,
Crisostomo,A., de Guia,R., Farrow,F.,
Pena,G., Sarile,A., Torres, P. (2014).
Laboratory Manual in Organic Chemistry.
Quezon City, Philippines: C & E Publishing.