Quantitative Inorganic Analysis Laboratory

Formal Report
Date Performed: 6
November 2015
Date Submitted: 20
November 2015
Department of Mining,
Metallurgical and
Materials Engineering
College of Engineering
Instructor: Searle
Aichelle Duay
Spectrophotometric Determination of the Acid
Dissociation Constant of Methyl Red
D.A. Mercado
The acid dissociation constant of methyl red was obtained by using the
spectrophotometric method. The maximum wavelengths were determined from both the
acidic and basic forms of the methyl red and were used in obtaining the absorbance. In
order to determine the molar absorptivity constant, the sample solutions were prepared
by mixing the specified values of HCl and HMR as well as a mixture of NaOAc and
MR-. The acid dissociation constant was then determined by measuring the absorbance
of a mixture prepared by adding HOAc and NaOAc into the standard methyl red
solution. This then lead to the calculation of the acid dissociation constant at 1.3142 x
10-5 which had a percent error of 31.42% from the theoretical value of 1.00 x 10 -5. [1]

Introduction This experiment aims to determine the acid

dissociation constant of methyl red by using a
Methyl red, also known as 4- spectrophotometric method on a multi-component
dimethylaminobenzene-2-carboxylic acid, is system.
generally used as the indicator in acid-base titrations.
It can exist in either an acidic (HMR) or basic (MR -) Materials and Methods
form which can be seen in the equation,
The sample solutions were prepared according to
HMR MR- + H+ Table 1 with solutions 1-6 done in a 50 mL volumetric
(1) flask (not diluted to mark) and 7-10 in a 100 mL beaker.
They were then swirled in order to properly mix all the
This equation then gives an equilibrium constant, K, solutions.
as seen in equation 2 and can then be expressed in
terms of pH by using the Henderson-Hasselbalch Table 1. Preparation of Sample Solutions
equation. [2] Solution Volume of 0.01 M Volume of Solution
Number HCl HMR
[ MR ][ H ] 1 4.96 mL 15.04 mL
K= 2 10.00 mL 10.00 mL
[ HMR]
(2) 3 15.04 mL 4.96 mL
Solution Volume of 0.01 M Volume of Solution
[ MR ] Number NaOAc MR-
pH pK a log (3) 4 4.96 mL 15.04 mL
[ HMR]
5 10.00 mL 10.00 mL
Since both the acidic and basic forms can absorb in 6 15.04 mL 4.96 mL
the visible region of light, spectrophotometry can be Solution Volume of Volume of Volume of
used in order to determine its acid dissociation Number Standard 0.02 M 0.04 M
constant. Due to the multi-component system of the Methyl Red HOAc NAOAc
solution, the total absorbance was determined by 7 6.00 mL 1.20 mL 12.80 mL
using the equation, 8 6.00 mL 2.40 mL 11.60 mL
9 6.00 mL 4.80 mL 9.20 mL
Atotal = A1 + A2 + +An = 1bc1 + 2bc2 + + nbcn (4)
10 6.00 mL 7.20 mL 6.80 mL
Where in the (A) signifies the absorbance, the ()
represents the molar absorptivity, the (b) gives the The spectra of the HMR and MR - solutions were
path length and the (c) denotes the concentration. then measured between 350 and 600 nm with H 2O in the
reference cell. This was then used in order to determine
the wavelength of maximum absorption of the two
Spectrophotometric Determination of the Acid Dissociation Constant of Methyl Red Mercado, 2015| 1
solutions. The molar absorptivity constant of HMR and constants, , were then given by the slope of the
MR- was then determined by measuring the absorbance equations of the linear lines and are seen in Table 2.
of solutions 1-10 at HMR and MR- with water in the
reference cell. The pH of solutions 7-10 were then Figure 1. Absorbance vs. Concentration
obtained using a pH meter in order to determine the acid
dissociation constant of HMR

Results and Discussion

Methyl red is a weak organic acid that can take an
acidic and basic form and is commonly used as a titration
indicator. Its acidic form is given by HMR which exhibits
a pink color while its basic form is seen as a yellow MR -
which is allows both solutions to absorb light and can
thus be analyzed through spectrophotometry.

In the preparation of the solutions, it was made sure that

the HMR had a pH of ~2 using 0.100 M HCl to ensure
that the methyl red is present only in the form of HMR.
Similarly, the MR- was kept at pH ~ 8 with 0.04 M
NaOAc since at that pH level, only the basic form of Table 2. Molar Absorptivity Constant
methyl red is present in the solution. Their maximum HMR at HMR MR- at HMR [L HMR at MR- [L MR- at MR-
absorbances were then measured and were found to be [L mol-1 cm-1] mol-1 cm-1] mol-1 cm-1] [L mol-1 cm-1]
520 nm and 428 nm for HMR and MR - respectively. The 39752 1282.3 3633.3 16670
absorbances of the calibration sample solutions 1-6 were
then measured at these wavelengths and can be seen in
These molar absorptivity constant values were then
Table 1 using matched cells. Matched cells are cuvettes
used in determining the concentrations of HMR and MR -
that are identical that are used in order to ensure that the
in solutions 7-10 using the following equations that were
absorbance subtracted from the reference cell containing
derived from equation (4).
water would be constant. [7]

Table 1. Absorbances and Molarities of Solutions 1-6 AHMR ( HMR ,HMR )b(c HMR ) ( MR ,HMR )b(c MR )
Solution Absorbance Molarity [M]
HMR- [nm] MR- [nm] (6)
1 0.498 0.037 1.3962 x 10 AMR ( HMR ,MR )b(c HMR ) ( MR ,MR )b(c MR )
5
(7)
2 0.313 0.022 9.283 x 10 6
3 0.126 0.003 4.604 x 10 6 Equations (6) and (7) can then be further manipulated to
give equations (8) and (9) which then lead to the values
4 0.014 0.220 1.3962 x 10 calculated in Table 3.
5

5 0.008 0.142 9.283 x 10 6 ( AMR MR,HMR ) ( AHMR MR ,MR )

6 0.002 0.064 4.604 x 10 6 [HMR]=
b( HMR ,MR MR ,HMR MR,MR HMR ,HMR )
(8)
AHMR HMR ,HMR bc HMR
The molar absorptivity constants were then determined [MR-]=
using the Beer-Lamberts Law in equation 5 by plotting b MR ,HMR
the absorbance measured against the concentration (9)
A = bc (5) Table 3. Absorbances, Molarities and pH of
Solutions 7-10
This then formed four lines which consisted of [HMR] Absorbance
vs. HMR , [HMR] vs. MR- , [MR-] vs. HMR and [MR-] vs. Soln pH [HMR] [MR-]
HMR MR-
MR- which is given by Figure 1. The Molar absorptivity
7 6.64 0.117 0.872 1.264 x 10 6 5.2056 x 10 5
Spectrophotometric Determination of the Acid Dissociation Constant of Methyl Red Mercado, 2015| 2
8 6.38 0.181 0.937 2.954 x 10 6 4.9575 x 10 5 from the nature of the solvent, errors in obtaining the pH
and preparing the solutions and mishandling of the
9 5.73 0.338 0.785 7.033 x 10 6 4.556 x 10 5 cuvettes and instrument which could lead to the deviation
of the experimental value from the theoretical. [3]
1.2507 x 10
10 5.46 0.549 0.719 5 4.0412 x 10 5
It was important to measure the pH of solutions 7-10 so
that the Henderson-Hasslebalch equation in (3) can be Conclusions and Recommendations
used in order to determine the acid dissociation constant
of methyl red by plotting log [MR-]/[HMR] against the The acid dissociation constant of methyl red was
pH. found to be 1.3142 x 10-5 which had a relative error of
31.42 % from the theoretical value of 1.00 x 10-5. [1] The
Figure 2. pH vs. log [MR-]/[HMR] molar absorptivity constants of HMR at HMR (520 nm)
and MR- (428 nm) were found to be 39752 L mol -1 cm -1
and 3633.25 L mol -1 cm -1, respectively. The molar
absorptivity constants for MR- at at HMR and MR- were
also found to be 1282.33 L mol -1 cm -1 and 16670 L mol
-1
cm -1 in that order. Alternative methods that could be
used other than spectrophotometry would be colorimetry,
potentiometry and liquid chromatography. [6]
References

[1] Quantitative Inorganic Analysis Manual; Institute of

Chemistry: UP Diliman, Quezon City, 2007.

[2] Colby College. www.colby.edu/chemistry/PChem/

The acid dissociation constant pK a was obtained through lab/pKaMethylRed.pdf (accessed 10 November 2015).
the y-intercept of the linear equation of the plot in Figure
2 giving a theoretical value of 4.88 which yielded a [3] Skoog, D.; West, D.; Holler, F.J.; Crouch, S.
relative error of 2.4% from the literature value of 5.00. [1] Fundamentals of Analytical Chemistry,9th ed.; Cengage
The Ka was then calculated to be 1.3142 x 10 -5 which Learning Asia Pte Ltd: Singapore, 2014.
resulted in a relative error of 31.42% from the actual
value of 1.00 x 10-5. The deviations in the experimental [4] Petrucci R.H. General Chemistry: Principles and
from the theoretical value could have been caused by the Modern Applications. Prentice Hall Inc.: Upper Saddle
limitations of Beer-Lamberts Law, one of which is the River, New Jersey 1997.
real limitation where in the analyte should be very dilute
and lower than 0.01 M. When the analyte has a high [5] University of California, Davis. chemwiki.
concentration, its individual particles start to affect one ucdavis.edu/Analytical_Chemistry/Analytical_Chemistry
another and are no longer independent thus causing a _2.0/10_Spectroscopic_Methods/10B
change in its absorbance. The absorptivity constant also %3A_Spectroscopy_Based_on_Absorption#10B.5_Limit
depends on the analytes refractive index. The ations_to_Beer.E2.80.99s_Law (Accessed November 9,
absorptivity may change when the concentration is high 2015).
but remains constant when it is dilute. Beers Law also
has chemical limitations where in the analyte must not [6] Whitten, K.; Davis, R.; Peck, M.L.; Stanley, G.
undergo an equilibrium reaction while being analyzed. It Chemistry, 9th ed.; Brooks/Cole, Cengage Learning:
may cause a change in color and would result to a change Canada, 2010.
in absorbance as well. Lastly, Beers Law is subject to
instrumental limitations since it makes an assumption
that the radiation that hits the sample is monochromatic
or of a single wavelength. A polychromatic radiation
would result in a negative difference from Beers Law. A
stray radiation may also cause deviations from the actual
value which is caused by any defect in the machines
wavelength selector. [5] Other sources of error could be
Spectrophotometric Determination of the Acid Dissociation Constant of Methyl Red Mercado, 2015| 3
 Appendix

Graph

Figure 1. Absorbance vs. Concentration

Spectrophotometric Determination of the Acid Dissociation Constant of Methyl Red Mercado, 2015| 4
 Figure 2. pH vs. log [MR-]/[HMR]
Calculations

Molarity of Solutions
(0.05 g )
Mstock = 0.00371333M
( 269.3 g / mol )(0.05 L)
Solution 1 and 4
2.5mL 5mL 15.04mL 5
0.0037133 M 1.3962 x10 M
50 mL 50mL 20 mL
Solution 2 and 5
2.5mL 5mL 10.00mL 6
0.0037133M 9.283x10 M
50mL 50mL 20mL
Solution 3 and 6
2.5mL 5mL 4.96mL 6
0.0037133M 4.604 x10 M
50mL 50mL 20mL
Solution 7
(0.872)(1282.33) (0.117 )(16670)
[HMR]= 1.264 x10 6 M
1[(3633.25)(1282.33) (16670)(39752)]
(0.117 ) (39752)(1)(1.264 x10 6 )
[MR ]= 5.2056 x10 5 M
(1)(1282.33)
Solution 8
(0.837)(1282.33) (0.181)(16670)
[HMR]= 2.954 x10 6 M
1[(3633.25)(1282.33) (16670)(39752)]
(0.181) (39752)(1)( 2.954 x10 6 )

[MR ]= 4.9575 x10 5 M
(1)(1282.33)
Solution 9
(0.785)(1282.33) (0.338)(16670)
[HMR]= 7.033x10 6 M
1[(3633.25)(1282.33) (16670)(39752)]
(0.338) (39752)(1)(7.033x10 6 )
[MR ]= 4.556 x10 5 M
(1)(1282.33)
Solution 10
(0.719)(1282.33) (0.549)(16670)
[HMR]= 1.2507 x10 5 M
1[(3633.25)(1282.33) (16670)(39752)]
(0.549) (39752)(1)(1.2507 x10 5 )
[MR ]= 4.0412 x10 5 M
(1)(1282.33)
Acid Dissociation Constant
10 4.881322016 1.3142 x10 5
Relative Error
(1.00 x10 5 ) (1.3142 x10 5 )
x100 31.42%
1.00 x10 5

Spectrophotometric Determination of the Acid Dissociation Constant of Methyl Red Mercado, 2015| 5