Quantitative Inorganic Analysis Laboratory

Formal Report

Date Performed: 21 April

2016 Spectrophotometric Determination of the
Date Submitted: 05 May
2016
Institution/Department:
Acid Dissociation Constant of Methyl Red
Department of D. B. Aguto
Mining, Metallurgical, and
Materials Engineering This experiment aimed to use a spectrophotometric method in determining the
College: Engineering acid dissociation constant of methyl red. The concentrations of HMR and MR - in
Instructor: Michelin the unknown was determined using Beer’s Law, which were then used in the
Ang computation of the pKa by the use of the Henderson-Hasselbach equation. The
experimental acid dissociation constant was calculated at 4.24, with a 14.34%
error from the literature value of 4.95.
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 𝐴 𝜆1 = 𝜀 𝐻𝑀𝑅,1𝑏𝑐 𝐻𝑀𝑅 + 𝜀 𝑀𝑅−,𝜆1𝑏𝑐 𝑀𝑅− [2]
EM radiation. 1 Molecular species absorb a specific at λ2 (λMR-) :
frequency of EM radiation. This causes the transfer of 𝐴 𝜆2 = 𝜀 𝐻𝑀𝑅,2𝑏𝑐 𝐻𝑀𝑅 + 𝜀 𝑀𝑅−,𝜆2𝑏𝑐 𝑀𝑅− [3]
energy from the radiation to the molecule, exhibiting
a transmittance of lesser intensity. Double component spectrophotometry was used in
Spectrophotometric methods use this principle to this experiment to determine the acid dissociation
determine multiple properties of a species such as its constant of methyl red, by first determining [HMR]
concentration and 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
determined using spectrophotometric methods Preparation of solutions. The following solutions
because both its acid (HMR) and basic (MR -) natures were prepared quantitatively: 50 mL methyl red stock
have strong absorption peaks in the visible portion of solution, 50 mL methyl red standard solution, 50 mL
the spectrum. Its acid dissociation constant is not 0.010 M NaCH 3COO, 50 mL 0.020 M CH 3COOH, 50
influenced by the changes in ionic strength, and the mL 0.010 M HCl, 50 mL working methyl red
color change between pH 4-6 can be conveniently standard solutions (HMR and MR -), as well as the
obtained using a simple acetate buffer system. 2 standard and sample solutions.
The ratio [MR-]/[HMR] may be determined Determination of λHMR and λMR-. The spectra of both
spectrophotometrically because both forms of methyl solutions were obtained between 350-600 nm using
red absorb strongly in the visible spectrum. Two water in the reference cell. Matched cells were used
wavelengths are selected for analyzing mixtures of for all absorbance measurements. The wavelengths
the two forms so that at one, the acidic form has a λHMR and λMR- were determined from the obtained
very large absorbency index compared with the basic spectra.
form, and at the other, the situation is reversed. The Absorbance and pH measurements of standard
absorbency indices of HMR and MR - are determined and sample solutions. The absorbance of solutions 1-
at both wavelengths, using several concentrations to 10 were measured at both λ HMR and λMR- using water
determine whether or not Beer’s law is obeyed. in 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
ARTICLE Journal Name

In the first part, methyl red standard solutions were at λHMR and λMR versus concentration
prepared with the same amount of methyl red in both
[HMR] and [MR-] sample solution. The absorptivity (𝜀) of the two forms of methyl red in
For the [HMR] solution, methyl red solution was both wavelengths was determined using Beer’s law.
acidified with dilute HCl solution at near pH 2.0. The
methyl red solution was prepared at this pH to convert Table II. Calculated Molar Absorptivity of HMR and MR
all MR- to HMR. For the [MR-] solution, it was basified solutions
with dilute NaOH solution at near pH 8.0. Similarly, the Species ε in HMR ε in MR
methyl red solution was prepared at this pH to convert HMR 3.47 x 107 3.56 x 106
all HMR to MR-. The pH of the solutions was measured MR- 1.19 x 106 1.70 x 107
and verified using the pH meter. These solutions were
then used in determining the spectra in which the acid
A series of solutions was prepared by adding varying
and basic forms of methyl red absorb the most.
amounts of dilute acetic acid to constant amounts of standard
Absorbance was measured between 350-600 nm
indicator solution buffered with dilute NaOAc solution. The
using the UV-Vis Double Beam Spectrophotometer.
absorbances (AλHMR and AλMR-) of the solutions at λ HMR and
The absorption peak for [HMR], λHMR, is at 520.4 nm,
λMR- were measured, and the pH values were determined
while the absorption peak for [MR -], λMR-, is at 428.4
using a pH meter. The exact pH of the solution will give
nm. These were used in determining the absorbance of
a picture of the ratio of the two forms of methyl red in
the sample solutions throughout the experiment.
the solution.
The absorbance of solutions 1-10 is measured at
The concentrations of HMR and MR - in the sample
these two wavelengths.
solutions were determined using equations [1] and [2].
Table I. Measured absorbance of solutions
Table III. Calculated concentration of HMR and MR in the
Solutio Absorbance
solutions
n λHMR λMR
Solution [HMR] [MR]
Number
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
6 .018 .061 To determine the experimental pKa value, pH was
plotted against log([MR]/[HMR]). This is the reason
7 .166 .919
why the pH of the solution was measured prior the
8 .279 .879
calculations.
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
follows.

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

-
Fig 1. Absorbance of HMR and MR

2 | J. Name., 2012, 00, 1-3 This journal is © The Royal Society of Chemistry 2012
Journal Name ARTICLE

K=¿ ¿ [2]
pH= pK + log¿ ¿ [3]

In the graph above, the y-intercept or the pKa is References

4.24. This gives a 14.34% error from methyl red’s
theoretical acid dissociation constant of 4.95. 4 Possible [1] Skoog, D.A. Fundamentals of Analytical Chemistry,
errors that could have contributed to this deviation are Ninth Edition. Holt, Rinehart and Winston.
as follows: the use of a mismatched cuvette, bubbles California, 2014.
trapped inside of cell, and the wrong addition of
samples. [2] Tobey, S.W. The Acid Dissociation Constant of
Methyl Red. A Spectrophotometric Measurement.
Conclusions and Recommendations Journal of Chemical Education [Online] 1958, 514,
Vol. 35, No. 10.
The experimental acid dissociation constant of
methyl red is found to be 4.24, with 14.34% error from [3] Quantitative Inorganic Analysis Laboratory Manual.
the literature value of 4.95.
2007 Edition.
The use of spectrophotometric methods in the
determination of the acid dissociation constant of
methyl red is reliable, despite encountering errors [4] Streuli, C.A. Handbook of Analytical Chemistry.
during the execution of the experiment. This method has McGraw-Hill. New York, 1963.
always been used for such purposes. In this experiment,
the right spectra should always be used in determining [5] Chemistry 111: Introduction to Spectrophotometry.
the wavelength of peak absorbance and the solutions employees.oneonta.edu/kotzjc/LAB/Spec_intro.pdf.
should always be diluted. Calibration of the instrument (accessed 04 May 2016)
should always be done before the experiment to make
sure that the instrument is sound and capable of
reporting reliable measurements.

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