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EXPERIMENT 6

ACID DISSOCIATION CONSTANT OF METHYL RED


A Laboratory Report in Partial Fulfilment of the Requirements in Chem117 Laboratory

DAGONDON, VANESSA OLGA


CAGAMPAN, JOHN SULUMOR
(Group 1, Cluster 1)
With
Blyth Angela Balgos
Christine Debbie Shanne Angeles
(Group 2, Cluster 1)

Chem117 Laboratory Physical Chemistry II


Section 1

Performed on March 24, 2016


Submitted on May 3, 2016

Mr. Arnold Gaje, RCh


Laboratory Instructor

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ABSTRACT
The experiment aims to determine the acid dissociation constant of methyl red through
spectrophotometric measurements based on Beer-Lamberts Law for mixtures and through the
Henderson Hasselbach equation. The absorbances were obtained using a UV VIS
spectrophotometer and the pHs were obtained using a pH meter. Two standards were prepared:
the acidic and basic form of methyl red. The wavelength at the highest absorbance was
determined for highest concentration of the two standards. After, the absorbances of the two sets
of standards were recorded. Four external calibration curves were generated by plotting the
absorbance against the concentration for the two standards at two wavelengths. Through these,
the concentration of the HMR and MR in each sample was calculated. Using the Henderson
Hasselbach equation the pKa of methyl red was obtained in each sample. The average pKa was
determined to be 4.86 0.03. This value obtained 3.82% relative error when compared to a
literature value of 5.05 0.05 determined by number of workers (Tobey, 1958).

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INTRODUCTION
Methyl red, a dimethylaminobenzene-2-caboxylic acid exist as a zwitter ion in aqueous
solutions. In acid solutions methyl red exist in the form of HMR. When base is added, a proton is
lost and methyl red exist as the yellow anion MR -. The equilibrium constant for the ionization of
methyl red is:
MR

pK = pH - log

(1)

In which the ionization constant at known pH values may be calculated from the measurements
of the ratio of (MR-)/(HMR).

Fig.1. HMR and MR- forms of methyl red (Tobey, 1958).


Spectrophotometry can be used to determine the ratio of (MR-)/(HMR) since the two
forms of methyl red absorb strongly in the visible range. Once the absorption spectra of the
methyl red in both acidic and basic solutions is determined using a spectrophotometric
instrument, two wavelengths are selected for analyzing mixtures of the two forms. The two
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wavelengths serves as an indication at which the acidic form of methyl red can have a very large
absorbancy index compared to the basic form at the first wavelength, and the reversed situation
at the other wavelength, this processed helps to assess if the using several concentrations of
methyl red can determine if Beers law is obeyed (Bell, 1958).
Beer-lamberts law states that the total absorbance of a solution at any given wavelength
is equal to the sum of the individual components of the solutions. Thus, the composition of a
mixture of HMR and MR- may be calculated from the absorbance of A1 and A2 at the two
wavelengths using:
A1 = 1,HMR(HMR) + 1,MR-(MR-)

(2)

A2 = 2,HMR(HMR) + 2,MR-(MR-)

(3)

Where there are two species present in the solution. The absorption spectra of each species tends
to overlap.
In the experiment the objective is to prepare and measure the visible absorption spectra of
acidic and basic forms of methyl red, thereby determining the wavelength of maximum
absorbance of the two solutions, then to create a calibration curve for each of the solutions and
molar absortivities at maz, to determine the relative amounts of acid and it conjugate base in
solutions. And ultimately to determine the pKa or the acid dissociation constants by following
the change in absorbance as a function of pH.

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METHODOLOGY
Preparation of solutions.
The stock solution of methyl red was prepared by dissolving a 1g of crystalline methyl
red (MR) in 300 mL of 95% ethanol and then diluting it to 500 mL with distilled water. A
standard solution of MR was used in the actual experiment, it was prepared by taking out 4 mL
of the stock solution and adding it to 50 mL of 95% ethanol and was diluted to 100 mL with
addition of water. A set of standard solutions was prepared. A 250 mL of 0.04 M NaOAc, 100 mL
of 0.01 M NaOAc, 100 mL of 0.02 M HOAc, 25mL of 0.1 M HCl, and 100 mL of 0.01 M HCl.
Two set solutions was prepared. Solution A was prepared by adding a 10 mL MR
solution with 10mL of 0.1M HCl into a 100 mL volumetric flask and was diluted to 100 mL with
addition of water. Solution B was prepared by adding 10 mL of MR solution with 25 mL 0.04M
NaOAc and was diluted to 100 mL in a 100mL volumetric flask with water. Each of the solution
was diluted into 0.75, 0.5, 0.25 %Concentration of their original concentration by using 0.1 M
HCl and 0.04M NaOAc in a 100 mL volumetric flask.
Determining the wavelength of maximum absorbance.
Solutions A and B was placed in a cuvette and the absorbance of solution was measured
between 350-600 nm to measure their maximum absorbance. And the absorbance was plotted for
the two solutions and XA and XB was aslo determined.
A series of solutions was prepared (solutions 1to 4) by adding varying amounts of 0.02 M
acetic acid, 50 mL, 25mL, 10 mL and 5mL respectively to 10 mL of standard indicator solution

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buffered plus 25 mL of 0.04 M NaOAc solution and diluting the solutions with water up to 100
mL of the volumetric flask.
Determining the molar absorptivities of [HMR] and [MR-] at max.
The absorbance of the solutions from set A and set B was placed in a cuvette separately
and the absorbance of every solution was determined using a UV-Vis. The absorbance was
plotted against the concentration and the molar absortivities of [HMR] and [MR-] was
determined using the equation
A=bc

Where

A
bc

(4)

(5)

A = absorbance
b = path length
c = concentration
= molar absorptivity

Determining the relative amounts of acid and its conjugate base.


The pH level of the solutions 1 to 4 was determined using a pH meter to ensure it doesnt
reach a basic level. The relative amounts of acid and its conjugate base was also determing by

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measuring the absorbance of the solutions at different wavelength with maximum absorbance
determined from the solutions A and B (Daniels et. al, 1970).
RESULTS
The following results reflect consecutively each of the objectives stated in order to
achieve the ultimate goal of the experiment, to be able to determine the equilibrium constant for
the ionization of methyl red, Ka, using spectroscopic measurements. The average value of Ka
expressed as a p value was determined to be 4.86 0.03. This value obtained 3.82% relative
error when compared to a literature value of 5.05 0.05 determined by number of workers
(Tobey, 1958).
Figure 1 shows the absorption spectra of the two solutions: A (acidic solution of Methyl
Red, HMR) and B (basic solution of Methyl Red, MR -). The maximum wavelength for solution
A is 520 nm and the maximum wavelength for the solution B is 428.5 nm.

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0.8
0.7
0.6
0.5
Absorbance

0.4
HMR
0.3

MR

0.2
0.1
0
350

400

450

500

550

600

650

Wavelength (nm)

Figure 1. Absorption spectra: Absorbances of HMR and MR- versus


Absorbances at the two selected maximum wavelengths were measured for the two sets
of standards prepared. Each of the standards consists of different concentrations of HMR and
MR-: 0.25, 0.5, 0.75, and 1 ppm. The recorded absorbances measured at each wavelength were
plotted against the concentrations of the standard. Figure 1 show four linear plots of absorbance
against wavelength at 520 nm and 428.5 nm for the two standards solutions.

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0.8
0.7
f(x) = 0.69x
0.6
HMR @ 428.5nm

Linear (HMR @ 428.5nm)

MR @ 428.5nm

HMR @ 520nm

Linear (HMR @ 520nm)

0.5

Absorbance

0.4

Linear (MR @ 428.5nm)


0.3

f(x) = 0.29x
0.2
0.1520nm)
Linear (HMR @
0
0.2

MR @ 520nm

f(x) = 0.05x
f(x) = 0.03x
0.3
0.4

0.5

Linear (MR @ 520nm)

0.6

0.7

0.8

0.9

1.1

Relative Concentration of Methyl Red

Figure 2. Absorbances of HMR and MR- at 520 and 428.5 versus concentration
Four samples of unknown solution were prepared. The absorbances of these solutions
were measured at the two selected wavelengths. The pHs of the samples were also obtained
using a pH meter. Table 1 shows the summary of the experimental data obtained for each of the
samples.
Table 1. Experimental data for the samples
Volume of

Absorbance at

Absorbance at

Solution
1
2

pH
HOAc (mL)

520

428.5

50
25

0.66
0.578

0.1
0.124

4.06
4.37
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3
4

10
5

0.273
0.148

0.234
0.27

5.19
5.6

The slope of the linear plots at Figure 2 denotes the absorptivity of each solution at each
wavelength. These values are used to calculate for the concentrations of MR - and HMR via
Beers Law. Using the concentrations of MR- and HMR and the measured pH of each the
samples, the pKa can be determined. Table 2 shows summary of the results for each of the
samples.
Table 2. Summary of Result for each sample

Solution

[HMR]

[MR-]

pKa

0.948

0.167

4.81

0.825

0.275

4.85

0.365

0.747

4.88

0.177

0.908

4.89

The pKa values obtained in Table 3 is averaged to be 4.86 0.03.


DISCUSSION
The acid dissociation constant of Methyl red (MR) was determined using
spectrophotometric measurements. Methyl red exists as zwitterion in aqueous solutions and has
resonance structure between its acidic (HMR) and basic form (MR -). It was observed that its
acidic form is pink in color while its basic form is yellow in color and thus both will have strong
absorption peaks in the visible portion of the spectrum (Toby, 1958). The acid-dissociation
constant can be calculated using:
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MR

K=

(6)

A more useful in determining the acid-dissociation constant is the Hender Hasselbach


shown in eq. 1. To determine the acid dissociation constant K, the pH and the concentrations of
the acidic and basic form of the MR must be known. In the experiment, the relative
concentrations of HMR and MR- were determined using spectrophotometric measurements and
the pH of the sample was determined using a pH meter.
Four samples of methyl red solutions were analyzed. Two sets of standards were
prepared: HMR and MR-. For each set of standard, calibration curves were generated by plotting
the absorbances of the standards against the relative concentration. A UV-Vis spectrometer as
used to determine the absorbances of the solutions. Before reading the absorbances of the
standards and sample, the maximum wavelength at which these absorbances will be read was
determined. Figure 1 shows the absorption spectra of the HMR and MR -. The absorption
spectrum is a plot of the absorbance against the wavelength. The wavelength with highest
recorded absorbance was selected both HMR and MR-. This was done to improve the sensitivity
of the signal measured that is the absorbance can still be measure effectively with small changes
in the concentration (Skoog et. al, 2000). The absorbances of the standards were measured at the
two wavelengths selected. Figure 2 shows the four calibration curves generated from the two sets
of standards.
The principle behind this spectrophotometric determination of the concentrations of
HMR and MR- is the beers law. Beers law states that the absorbance is equal to the
concentration, pathlength and molar absorptivity. There is a linear relationship between the
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absorbance and the concentration. Four calibration curves were generated by plotting absorbance
against the concentration in each of HMR and MR- as shown in figure 2. The slope represents
that absorptivity since the path length is 1 cm. The slope and the equation of the best fitted in
each of the linear plots was determined using the least square methods. Beers law can also be
applied in mixtures. Absorbance is additive and if the pathlength b is equal to 1 cm, it can be
expressed as:

MR ( )

A =[HMR] ( )HMR +
520

520

(7)

520

MR ( )

=[ HMR]( ) HMR +
428.5

428.5

(8)

428.5

Since the absorbance of the sample is known, together with the Absorptivities which are
just the slopes of the calibration curves, the values for [HMR] and [MR -] can be calculated.
Moreover, the pH of the sample was determined using a pH meter. Table 1 summarizes the pH
readings of each of the sample prepared. Using the Henderson Hasselbach equation, the pKas
of the samples were calculated and summarized in Table 2. The average of the obtained pKa
values is 4.86 0.03. This resulted to an error of 3.82% relative error when compared to a
literature value of 5.05 0.05 determined by number of workers (Tobey, 1958).
This minimum value of relative error indicates that there is no systematic error in the
analysis just random errors. There random errors might originate from the preparation of the
standards, the absorbance readings, the fluctuation of temperature and from personal errors.
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CONCLUSION
Methyl red is a pH sensitive azo dye used as indicator in volumetric titrations. In aqueous
solutions, methyl red exists as zwitterion with a resonance structure between its two forms: basic,
MR- and acidic, HMR. In acidic conditions, it forms a pink solution while in basic conditions, it
forms a yellow solution. Since these two forms absorb strongly in the visible range, the acid
dissociation constant of methyl red can determined using spectroscopic methods.
Four mixtures of HMR and MR were prepared to be analyzed. The pHs of the samples
were measured using the pH meter. Also, four calibration curves were prepared using the two
sets of standards, HMR and MR. The absorbances were read from a UV-VIS spectrophotometer
in the two selected wavelengths. Beers law is obeyed in the experiment. Beers law states that
the absorbance is equal to the absorptivity, path length and concentration. The path length of the
of the cuvette used in the UV Vis spectrophotometer is 1 cm. This makes the slope in the
calibration curve be equal to the absorptivity. The absorbance of the the mixture samples were
obtained. Since the absorbance is additive, two sets of equations were used to calculate the
concentration of HMR and MR in each sample. The acid dissociation constant was computed
using the Hneder Hasselbach equation. The average acid dissociation constant determined in
the experiment is 4.86 0.03 which obtained a 3.82% relative error when compared to a
literature value of 5.05 0.05 determined by number of workers (Tobey, 1958). Errors
considered in the experiment are random in nature. No systematic error has been detected and
therefore the experiment is a success.
REFERENCES
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Atkins, P.; de Paula, J. Physical Chemistry Ninth Edition; Oxford University Press: Great Britain,
2010.
Bell, C. K. Acids and Bases: Their Quantitative Behaviour; Methuen & Co., Ltd.: New York,
1952.
D.C. Harris, Quantitative Chemical Analysis, 7th ed. New York: W.H. Freeman and Company,
2007.
F. Daniels, J. W. Williams, P. Bender, R. A. Alberty, C. D. Cornwell, J. E. Harriman, Acid
Dissociation Constant of Methyl Red, Experimental Physical Chemistry, McGraw-Hill,
New York, NY, 1970, pp. 113-115.
Skoog, Douglas, et. al. Analytical Chemistry: An Introduction, Thomson Learning Asia. 2000.
Tobey, J. The Acid Dissociation Constant of Methyl Red. Journal of Chemical Education. 1958,
35, 514.
ACKNWLEDGEMENTS
We would like to express our deepest gratitude to our laboratory classmates and
especially to our cluster for working with us diligently, to our instructor for guiding us during the
experiment, and to the chemistry department for providing us the materials for the experiment.

CONTRIBUTION OF AUTHORS
Each of the members of the group contributed accordingly in the making of this
laboratory report: Cagampan authored the introduction and material and method sections;
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Dagondon, on the other hand, was in charge of the data analysis, the compilation of results,
discussion and also the conclusion; Cagampan wrote the abstract and proof read the overall
report.
APPENDICES
Appendix I. Tables and Figures
Table 1. Raw Data for the Absorbance and wavelength for HMR and MR standard solution
solution A (HMR)
wavelength absorbance
400
0.017
403
0.018
406
0.02
409
0.023
412
0.027
415
0.03
418
0.035
421
0.04
424
0.047
427
0.053
430
0.061
433
0.071
436
0.082
439
0.093
442
0.106
445
0.121
448
0.138
451
0.155
454
0.173
457
0.194
460
0.217
463
0.241
466
0.265
469
0.291
472
0.32
475
0.349
478
0.377
481
0.404

Solution B (MR-)
wavelength
absorbance
400
0.245
403
0.26
406
0.264
409
0.269
412
0.273
415
0.275
418
0.277
421
0.279
424
0.28
427
0.28
430
0.28
433
0.277
436
0.276
439
0.274
442
0.273
445
0.27
448
0.266
451
0.261
454
0.256
457
0.249
460
0.241
463
0.232
466
0.221
469
0.21
472
0.198
475
0.185
478
0.172
481
0.16
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484
487
490
493
496
499
502
505
508
511
514
517
520
523
526
529
532
535
538
541
544
547
550
553
556
559
562
565
568
571
574
577
580
583
586
589
592
595
598

0.436
0.466
0.494
0.52
0.55
0.577
0.602
0.624
0.643
0.66
0.67
0.677
0.679
0.676
0.671
0.663
0.652
0.64
0.626
0.61
0.594
0.569
0.541
0.509
0.471
0.425
0.373
0.327
0.278
0.228
0.184
0.145
0.114
0.086
0.063
0.047
0.035
0.025
0.019

484
487
490
493
496
499
502
505
508
511
514
517
520
523
526
529
532
535
538
541
544
547
550
553
556
559
562
565
568
571
574
577
580
583
586
589
592
595
598

0.144
0.131
0.118
0.106
0.093
0.081
0.071
0.063
0.055
0.046
0.04
0.035
0.031
0.026
0.023
0.021
0.018
0.016
0.015
0.014
0.013
0.012
0.012
0.011
0.011
0.01
0.01
0.01
0.009
0.008
0.007
0.007
0.006
0.006
0.006
0.005
0.005
0.005
0.005

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Table 2. Absorbances taken at the selected wavelengths for HMR and MR standards
Relative Concentration

MR-

HMR
520

428.5

520

428.5

0.25

0.139

0.012

0.01

0.073

0.5

0.334

0.028

0.014

0.16

0.75

0.526

0.04

0.023

0.208

0.7

0.056

0.026

0.283

Appendix B: Sample Calculations

Determination of absorptivity/slope of liner plot via least square method


For the linear plot of absorbance recorded at 520nm against relative concentration of HMR:

( x )2
6.25
S xx = x
=1.875
=0.3125
n
4
2

S yy = y 2

( y )2
2.8886601
=0.897553
=0.17590275
n
4

S xy =xy

m=

( xy )
4.2475
=1.29625
=0.234375
n
4

S xy 0.234375
=
=0.6913
S xx
0.3125

Determination of the concentration of HMR and MR in sample via algebraic method, substitution
For sample 1 only:
(520)HMR = 0.6913
(520)MR = 0.0281
(428.5)HMR = 0.0549
(428.5)MR = 0.2865

MR ( )

A =[HMR] ( )HMR +
520

520

520


MR ( )

=[ HMR]( ) HMR +
428.5

428.5

428.5

Solving for [MR] and [HMR]:

MR

MR ( ) MR

A
[ HMR] =

520

520

Determination of pKa
For sample 1 only:

MR

pK a= pH log