Quantitative Inorganic Analysis Laboratory

Formal Report

Spectrophotometric Determination of the Acid

Dissociation Constant of Methyl Red
Date Performed: 21 April
2016 D. B. Aguto
Date Submitted: 05 May
2016 This experiment aimed to use a spectrophotometric method in determining the
Institution/Department: acid dissociation constant of methyl red. The concentrations of HMR and MR- in
Department of Mining, the unknown was determined using Beer’s Law, which were then used in the com-
Metallurgical, and Materials
Engineering putation of the pKa by the use of the Henderson-Hasselbach equation. The exper-
College: Engineering imental acid dissociation constant was calculated at 4.24, with a 14.34% error
from the literature value of 4.95.
Instructor: Michelin Ang
Introduction The absorbance of the solution at the two peaks is
the sum of the absorbance of [HMR] and [MR-] at
Electromagnetic (EM) waves are composed of each wavelength.
small packets of energy called photons. The energy of at λ1 (λHMR) :
a photon is indirectly related to the wavelength of the 𝐴𝜆1 = 𝜀𝐻𝑀𝑅,1𝑏𝑐𝐻𝑀𝑅 + 𝜀𝑀𝑅−,𝜆1𝑏𝑐𝑀𝑅− [2]
EM radiation.1 Molecular species absorb a specific at λ2 (λMR-) :
frequency of EM radiation. This causes the transfer of 𝐴𝜆2 = 𝜀𝐻𝑀𝑅,2𝑏𝑐𝐻𝑀𝑅 + 𝜀𝑀𝑅−,𝜆2𝑏𝑐𝑀𝑅− [3]
energy from the radiation to the molecule, exhibiting
a transmittance of lesser intensity. Spectrophotomet- Double component spectrophotometry was used in
ric methods use this principle to determine multiple this experiment to determine the acid dissociation
properties of a species such as its concentration and constant of methyl red, by first determining [HMR]
its acid dissociation constant. and [MR-] in a solution where the two forms coexist.
Methyl red is a common indicator dye used in the From there, the Henderson – Hasselbach principle
qualitative analysis of acid-base reactions. It is red in was used to compute for the pKa.
pH under 4.4, yellow in pH over 6.2, and orange in
between. Materials and Methods
The dissociation constant of methyl red can be de-
termined using spectrophotometric methods because Preparation of solutions. The following solutions
both its acid (HMR) and basic (MR-) natures have were prepared quantitatively: 50 mL methyl red stock
strong absorption peaks in the visible portion of the solution, 50 mL methyl red standard solution, 50 mL
spectrum. Its acid dissociation constant is not influ- 0.010 M NaCH3COO, 50 mL 0.020 M CH3COOH, 50
enced by the changes in ionic strength, and the color mL 0.010 M HCl, 50 mL working methyl red stand-
change between pH 4-6 can be conveniently obtained ard solutions (HMR and MR-), as well as the standard
using a simple acetate buffer system.2 and sample solutions.
The ratio [MR-]/[HMR] may be determined spec- Determination of λHMR and λMR-. The spectra of
trophotometrically because both forms of methyl red both solutions were obtained between 350-600 nm
absorb strongly in the visible spectrum. Two wave- using water in the reference cell. Matched cells were
lengths are selected for analyzing mixtures of the two used for all absorbance measurements. The wave-
forms so that at one, the acidic form has a very large lengths λHMR and λMR- were determined from the ob-
absorbency index compared with the basic form, and tained spectra.
at the other, the situation is reversed. The absorbency Absorbance and pH measurements of standard and
indices of HMR and MR- are determined at both sample solutions. The absorbance of solutions 1-10
wavelengths, using several concentrations to deter- were measured at both λHMR and λMR- using water in
mine whether or not Beer’s law is obeyed. the reference cell. The pH of solutions 7-10 were
measured with a pH meter.
A = 𝜀𝑏c [1]
Results and Discussion

Spectrophotometric Determination of the Acid Dissociation Constant of Methyl Red Aguto, 2016| 1
 In the first part, methyl red standard solutions were
prepared with the same amount of methyl red in both The absorptivity (𝜀) of the two forms of methyl red in
[HMR] and [MR-] sample solution. both wavelengths was determined using Beer’s law.
For the [HMR] solution, methyl red solution was
acidified with dilute HCl solution at near pH 2.0. The Table II. Calculated Molar Absorptivity of HMR and MR
methyl red solution was prepared at this pH to convert solutions
all MR- to HMR. For the [MR-] solution, it was basified Species ε in HMR ε in MR
with dilute NaOH solution at near pH 8.0. Similarly, the HMR 3.47 x 107 3.56 x 106
methyl red solution was prepared at this pH to convert MR- 1.19 x 106 1.70 x 107
all HMR to MR-. The pH of the solutions was measured
and verified using the pH meter. These solutions were
A series of solutions was prepared by adding varying
then used in determining the spectra in which the acid
amounts of dilute acetic acid to constant amounts of standard
and basic forms of methyl red absorb the most.
indicator solution buffered with dilute NaOAc solution. The
Absorbance was measured between 350-600 nm us-
absorbances (AλHMR and AλMR-) of the solutions at λHMR
ing the UV-Vis Double Beam Spectrophotometer. The
and λMR- were measured, and the pH values were deter-
absorption peak for [HMR], λHMR, is at 520.4 nm, while
mined using a pH meter. The exact pH of the solution
the absorption peak for [MR-], λMR-, is at 428.4 nm.
will give a picture of the ratio of the two forms of me-
These were used in determining the absorbance of the
thyl red in the solution.
sample solutions throughout the experiment.
The concentrations of HMR and MR- in the sample
The absorbance of solutions 1-10 is measured at
solutions were determined using equations [1] and [2].
these two wavelengths.
Table III. Calculated concentration of HMR and MR in the
Table I. Measured absorbance of solutions
solutions
Solution Absorbance
Solution [HMR] [MR]
Number λHMR λMR
7 2.96065E-09 5.33494E-08
1 .497 .056
8 6.32065E-09 5.03074E-08
2 .387 .039
9 1.39794E-08 4.37074E-08
3 .175 .023
10 2.45516E-08 3.78582E-08
4 .029 .219
5 .019 .112
To determine the experimental pKa value, pH was
6 .018 .061
plotted against log([MR]/[HMR]). This is the reason
7 .166 .919
why the pH of the solution was measured prior the cal-
8 .279 .879
culations.
9 .537 .794
10 .897 .732
The y-intercept of the graph is the experimental pKa,
as derived from the equilibrium constant expression, K,
Using these absorbance measurements, the molar of methyl red.3
absorptivity of HMR and MR in both the λHMR and λMR
were identified using linear regression and it is as fol-
lows.

Fig. 2 log [MR-]/[HMR] versus pH

-
Fig 1. Absorbance of HMR and MR
at λHMR and λMR versus concentration [𝑀𝑅−][𝐻+]
𝐾= [𝐻𝑀𝑅]
[2]

Spectrophotometric Determination of the Acid Dissociation Constant of Methyl Red Aguto, 2016| 2
 [𝑀𝑅−]
𝑝𝐻 = 𝑝𝐾 + log ([𝐻𝑀𝑅])
In the graph above, the y-intercept or the pKa is
4.24. This gives a 14.34% error from methyl red’s theo-
retical acid dissociation constant of 4.95.4 Possible er-
rors that could have contributed to this deviation are as
follows: the use of a mismatched cuvette, bubbles
trapped inside of cell, and the wrong addition of sam-
ples.
Conclusions and Recommendations
The experimental acid dissociation constant of me-
thyl red is found to be 4.24, with 14.34% error from the
literature value of 4.95.
The use of spectrophotometric methods in the deter-
mination of the acid dissociation constant of methyl red
is reliable, despite encountering errors during the execu-
tion of the experiment. This method has always been
used for such purposes. In this experiment, the right
spectra should always be used in determining the wave-
length of peak absorbance and the solutions should al-
ways be diluted. Calibration of the instrument should
always be done before the experiment to make sure that
the instrument is sound and capable of reporting reliable
measurements.
Spectrophotometric Determination of the Acid Dissociation Constant of Methyl Red
[3]
References
[1] Skoog, D.A. Fundamentals of Analytical Chemistry,
Ninth Edition. Holt, Rinehart and Winston. Califor-
nia, 2014.
[2] Tobey, S.W. The Acid Dissociation Constant of Me-
thyl Red. A Spectrophotometric Measurement. Jour-
nal of Chemical Education [Online] 1958, 514, Vol.
35, No. 10.
[3] Quantitative Inorganic Analysis Laboratory Manual.
2007 Edition.
[4] Streuli, C.A. Handbook of Analytical Chemistry.
McGraw-Hill. New York, 1963.
[5] Chemistry 111: Introduction to Spectrophotometry.
employees.oneonta.edu/kotzjc/LAB/Spec_intro.pdf.
(accessed 04 May 2016)
Aguto, 2016| 3