Objective

The objective of the experiment was to measure the pKa value of the methyl red through its
absorbance spectra as a function of pH.

Introduction
In this experiment, we are using Methyl Red (4-dimethylaminobenzene-2-carboxylic
acid) which an acidic dye that will turn red in acidic solutions. It is an azo dye, and a dark red
crystalline powder. The objective of this experiment is to determine the pKa value of methyl
red solutions through its absorbance spectra as a function of pH. pKa is the inverse log of Ka
which Ka is the acid dissociation constant also known as acidity constant which is a
quantitative measures of the strength of an acid in a solution.
pKa = -log Ka
Ka = [A-][H+]/[HA]
In this experiment, the chemical reaction equation are as follows :
HMR + H2O

MR- + H3O+

Which Ka can be determined by :

[H3O+]
[ MR ] Ka =
-

[HMR]

And the pKa is :

[ MR-]
pKa = pH

log

[HMR]
The value of pKa can be determined by varying the value of pH of methyl red solutions and
calculate the value of [MR-]/[HMR].
In chemistry, absorbance or decadic absorbance is the common logarithm of the ratio
of incident to transmitted radiant power through a material, and spectral absorbance or

spectral decadic absorbance is the common logarithm of the ratio of incident to transmitted
spectral radiant power through a material. The BeerLambert law, also known as Beer's law,
the LambertBeer law, or the BeerLambertBouguer law relates the attenuation of light to
the properties of the material through which the light is traveling. The law is commonly
applied to chemical analysis measurements and used in understanding attenuation in physical
optics, for photons, neutrons or rarefied gases.
A= bc
Where, A = absorbance

molar absorptivity (L. cm-1. mol-1) b

length of the cell (cm) c

path

= concentration of the

absorbing species (mol. L-1)

The absorption spectrum of HMR and MR - is shown in Figure 1. The curve shows the
changes in the spectrum for the system with the changes in the pH. The absorbance values of
HMR and MR- cannot be differentiated if the molar extinction of both is similar at certain
wavelengths. The absorption for the overall system will be constant if the concentration for
both of the HMR and MR- is constant, regardless of the concentration ratio. This point is
known as the isosbestic point. This point is identified as all the curves are intersected.
(Figure 1)

Figure 1: Changes in the absorption spectrum of HMR and MR- with pH

Summary

In this experiment, we will measure pKa value of Methyl red solutions through its
absorbance spectra as a function of pH. We first prepare the methyl red solutions by adding
5ml of methyl red solution into a 100 ml of volumetric flask and then add 50 ml of ethanol
(95%) into the flask and dilute the solution using distilled water until it reaches the marked
level.
This experiment consist of two parts in which in part B we vary the solutions by
making two types of solution which is acidic and basic. For the basic solution we used
sodium hydroxide and then check the pH using pH meter and record the absorbance of the
solution. As for acidic solution we used hydrochloric acid and then check the pH using pH
meter and also record the absorbance. For part C, we varied the pH of the methyl red solution
by adding different volume of acetic acid and sodium acetate. These solutions then were
tested their pH and also recorded their wavelengths using spectrophotometer.

Materials
1.
2.
3.
4.
5.
6.

1000 ppm of Methyl Red Stock Solution

95% of Ethanol
0.1 M of Hydrochloric Acid, HCl (0.1 M)
0.1 M of Sodium Hydroxide, NaOH
0.2 M of Sodium Acetate
0.2 M of Acetic Acid

Equipment / Apparatus
1. Ultraviolet Visible Spectrophotometer with Sample cell (10 mm)
2. pH meter
3. Volumetric Flask (12 x 50 ml)
4. Pipette (1 ml, 5 ml and 10 ml)
5. Beaker (50 ml and 100 ml)
6. Dropper
Procedure:

Part A : Methyl Red Standard Solution

1.
2.

5 ml of methyl red stock solution is pipetted into 100 ml volumetric flask.

50 ml of ethanol (95%) is added into flask and diluted with distilled water until
mark level is achieved.

Part B : Methyl Red Acidic and Basic Solutions

i) Acidic Solution (methyl red)
1.
2.
3.

6 ml of methyl red of Part A is pipetted into 50 ml volumetric flask.

10 ml of hydrochloric acid (0.1 M) is added into the flask.
The solution in the flask is diluted with distilled water until mark level is

4.
5.
6.

achieved.
The pH of the solution is checked by using a pH meter.
The pH of the solution is labelled and its colour is observed.
The absorption spectrum for the solution in the wavelength range of 350-600 nm
is recorded.

ii)

Basic Solution (methyl red)

1.
2.
3.

5 ml of methyl red is pipetted into a 50 ml volumetric flask.

10 ml of sodium hydroxide (1.0 M) is added into the flask.
The solution in the flask is diluted with distilled water until mark level is

4.
5.
6.

achieved.
The pH of the solution is checked by using a pH meter.
The pH of the solution is labelled and its colour is observed.
The absorption spectrum for the solution in the wavelength range of 350-600 nm
is recorded.

Part C : Absorption Spectrum of Methyl Red

1.

A series of methyl red standard solutions is prepared in 50 ml volumetric flask and

labelled as A, B, C, D, E, F, G, and H.

Solution

Volume of MR standard

Volume of 0.2 M

Volume of 0.2 M

solution (ml)

acetic acid (ml)

sodium acetate (ml)

A
B
C
D
E
F
G
H

6
6
6
6
6
6
6
6

12
10
8
6
4
3
2
1

8
10
12
14
16
17
18
19

2.

The mixture in each volumetric flask is diluted with distilled water until the mark

3.

level
The volumetric flasks are shaken and the pH of the standard solutions is measured

4.

by using a pH meter.
The wavelength scan for all solutions including acidic and basic solutions of

5.

methyl red is performed using the Ultraviolet Visible Spectrophotometer.

The absorption spectrum for each solution is recorded on a different paper
immediately after the solutions are prepared.

Results
1)
Table 2: Absorbance of all solutions at various wavelength.

Graph 1 : Absorbance values against the wavelength.

Absorbance against Wavelength

1.2
1
0.8

Absorbance

0.6
0.4
0.2
0
360

410

460

510

Wavelength
Acidic

Basic

Isosbestic point.
=(465.9,0.43)
Maximum absorption wavelength left and right.

560

610

Left=(400,0.159)
Right=(520,1.036)
2) Absorbance value at maximum wavelength ( max)
Table 3 : pH and absorbance of all solutions
Solution

pH

Acidic
A
B
C
D
E
F
G
H
Basic

1.38
4
4.16
4.3
4.5
4.71
4.9
5.12
5.55
12.06

Absorbance at
max
1.036
0.807
0.752
0.663
0.555
0.45
0.465
0.49
0.519
0.378

Graph 2 : Absorbance (at maximum wavelength) against pH.

Absorbance at max against pH

1.2
1
0.8

Absorbance at max

0.6
0.4
0.2
0

10

12

pH

3) Concentration of [MR ] and [HMR] for each solution.

[MR ] = Abs at max of Solution X Abs at max of Basic Solution

14

[HMR] = Abs at max of Acidic Solution Abs at max of Solution

X
Solution

pH

Absorbanc
e at max

[MR ]

[HMR]

log[MR ]/
[HMR]

0.807

0.429

0.229

0.273

4.16

0.752

0.374

0.284

0.12

4.3

0.663

0.285

0.373

-0.117

4.5

0.555

0.177

0.481

-0.434

4.71

0.45

0.072

0.586

-0.911

4.9

0.465

0.087

0.571

-0.817

5.12

0.49

0.112

0.546

-0.688

5.55

0.519

0.141

0.517

-0.564

Graph 3 : Log [MR ] / [HMR] against pH of the solutions.

log[MR ]/[HMR] against pH

0.4
0.2

Log [MR ]/[HMR]

0
3.8
-0.2

= - 0.63x
4 f(x) 4.2
4.4 + 2.55
4.6

4.8

-0.4
-0.6
-0.8
-1

pH

CONCLUSION:

5.2

5.4

5.6

5.8

Based on this experiment, we manage to measure the pKa value of the methylred through its
absorbance spectra as a function of Ph. The values of pKa can be determined with using the
formula: pKa=Ph-log([MR-]/[HMR])
The overall lines of absorption by marking the intersection point of the similar level of
absorption from each pH value. This intersection point is called isosbestic point coordinated
at
(465.9, 0.43)
Based on the graph 3, log([MR-]/[HMR]) is inversely propotional to pH value. In graph 2, the
isosbestic point that we obtained when plotting the graph was at (465.9, 0.43) where 465.9
and 0.43 represented the wavelength and the absorption of spectrum respectively.
RECOMMENDATIONS:
There are a few precaution that should be taken when running this experiment. Firstly, make
sure to use stopper when using volumetric flask to avoid any impurities from getting in. we
also must rinse the pH meter with distilled water before measuring the pH value. In order to
avoid parallax error, make sure our eyes are perpendicular to the meniscus when taking
measurement.
LABORATORY QUESTION
Pre-lab Question
1.

Define pKa. Differentiate between pKa and pH.

-

pKa is the quantitative measure of an acid dissociation which negative logarithm of

the ionization constant (Ka) of an acid equal to the pH value with the present of equal
acid concentrations and conjugate bas forms. The difference between pH and pKa is
that pH measure the hydrogen ions in a certain medium while pKa we can state
degree which acid dissociates, preferably when the acid reach equilibrium .

2.

Describe about max.

-

max is when the absorbance of spectrum of solutions reached maximum which can
be observe by using spectrophotometer .

Post-lab Question

1. Discuss the straight line in the graph of log [MR-] / [HMR] against pH.
-As the value of pH solution increased the value log[MR-]/[HMR] decreased this can be tell
by referring the graph.
2. Calculate the pH of the prepared buffer solution (please use one example from any of the
solution). Compare the calculated pH value with the true pH that measure by pH meter.
pKa = pH + log [MR ]/ [HMR]
log [MR ]/ [HMR] = pKa pH
When log [MR ]/ [HMR] = 0
0 = pKa pH
pKa = pH
3. Discuss the weakness of the experiment
-Incorrect measurement of solutions , the spectrophotometer reading also can be
influenced if the cuvette didnt cleaned well .

REFERENCE
1. http://en.wikipedia.org/wiki/Isosbestic_point
2. http://en.wikipedia.org/wiki/Spectrophotometer
3. Physical chemistry lab manual