EXPERIMENT 6

Colorimetric
Determination of pH

DEL MUNDO
LARIN
SEE
INTRODUCTION
 Colorimetry
• any technique by which an
unknown color is evaluated in
terms of standard colors
• the technique may be visual,
photoelectric, or indirect by
means of spectrophotometry

 http://www.answers.com/topic/colorimetry#ixzz1MK1Yv3Sl
 pH Indicators
• also called acid-base indicators
• pH indicators are usually weak
acids or weak bases that change
their color depending on their
dissociation (protonation) state
• pH indicators can be used to
check pH of the solution

http://www.ph-meter.info/pH-measurements-indicators
 Indicator Lower pH pH Range Higher pH
color (transition color
interval)
Thymol blue Red 1.2 – 2.8 Yellow

Bromophenol blue Yellow 3.0 – 4.6 Purple

Chlorophenol red Yellow 4.8 – 6.4 Violet

Bromothymol blue Yellow 6.0 – 7.6 Blue

Phenol red Yellow 6.8 – 8.4 Red

 Buffer Solutions
• A buffer solution is one in which
the pH of the solution is "resistant"
to small additions of either a strong
acid or strong base. 
• Buffers consist of a weak acid and
its conjugate base or vice versa, in
relatively equal and "large"
quantities. 

http://www.chem.purdue.edu/gchelp/howtosolveit/equilibrium/buffers.htm
 McIlvaine Buffer System
• A citrate/phosphate buffer system
that can be volumetrically set for
pH in a wide range (2.2 to 8)

http://www.biochemlab.cn/shiji/peizhi/20993.html
 Henderson-Hasselbalch Equation
• An equation expressing the pH of
a buffer solution as a function of
the concentration of the weak acid
or base and the salt components
of the buffer.

http://medical-dictionary.thefreedictionary.com/Henderson-Hasselbalch+equation
 Colorimetric Analysis
• Uses the variation as a means of
determining the pH since the
intensity of the color of a solution
changes with its concentration or
pH

http://www.inc.bme.hu/en/subjects/genchem/phdet2.pdf
 • By comparing the intensity of the
colour of a solution of unknown
concentration (or pH) with the
intensities of solutions of known
concentrations (or pH), the
concentration of an unknown
solution may be determined

http://www.inc.bme.hu/en/subjects/genchem/phdet2.pdf
EXPERIMENT
Objective
• To be able to determine the pH of
an unknown solution
colorimetrically
• To be able to calculate the
ionization constant of a weak acid
PART A: Preparation of Buffer Solutions

A set of McIlvaine buffers were accurately

prepared in test tubes of uniform sizes labeled
according to their respective pH levels.

Five drops of the appropriate indicators to

use for each pH level were added to each of
the buffer solutions.
 Indicator: THYMOL BLUE (1.2 – 2.8)
pH 0.2M Na2HPO4 (mL) 0.1M Citric Acid (mL)

2.2 0.20 9.80

2.4 0.62 9.38

2.6 1.06 8.91

2.8 1.58 8.42

 Indicator: BROMOPHENOL BLUE (3.0 – 4.6)
pH 0.2M Na2HPO4 (mL) 0.1M Citric Acid (mL)
3.0 2.05 7.95
3.2 2.47 7.53
3.4 2.85 7.15
3.6 3.22 6.78
3.8 3.55 6.45
4.0 3.25 6.15
4.2 4.14 5.86
4.4 4.41 5.59
4.6 4.67 5.33
 Indicator: CHLOROPHENOL RED (4.8 – 6.4)
pH 0.2M Na2HPO4 (mL) 0.1M Citric Acid (mL)
4.8 4.93 5.07
5.0 5.15 4.85
5.2 5.20 4.80
5.4 5.58 4.42
5.6 5.80 4.20
5.8 6.05 3.95
6.0 6.31 3.69
6.2 6.61 3.39
6.4 6.92 3.08
 Indicator: BROMOTHYMOL BLUE (6.0 – 7.6)
pH 0.2M Na2HPO4 (mL) 0.1M Citric Acid (mL)
6.0 6.31 3.69
6.2 6.61 3.39
6.4 6.92 3.08
6.6 7.34 2.66
6.8 7.72 2.28
7.0 8.24 1.76
7.2 8.69 1.31
7.4 9.08 0.92
7.6 9.37 0.63
 Indicator: PHENOL RED (6.8 – 8.0)
pH 0.2M Na2HPO4 (mL) 0.1M Citric Acid (mL)
6.8 7.72 2.28
7.0 8.24 1.76
7.2 8.69 1.31
7.4 9.08 0.92
7.6 9.37 0.63
7.8 9.57 0.43
8.0 9.72 0.28
PART B: Colorimetric Determination of pH
 The pH of the following solutions were
approximated using pH papers, then applied them
with the appropriate indicator/s.

The pH of each solutions were then confirmed by

comparing their colors to standards (from part A)
applied with the same indicator.
Results
Solution Observed
pH
A 0.01M HOAc 5
B 1mL 0.1M HOAc + 1mL 0.1M NaOAc + 4.8
8mL H20
C 1mL 0.1M HOAc + 0.1mL 0.1M NaOAc 3.2
+ 8.9mL H2O
D 0.1mL 0.1M HOAc + 1mL 0.1M NaOAc 6.8
+ 8.9mL H2O
Discussion
• In the preparation of the buffer
solutions in part A, it is important to
use the appropriate indicators for
each buffer solutions because in
colorimetric determination of pH, the
indicators in buffered solutions are
most effective when it is within the
specific pH ranges mentioned in the
table. It is at these pH ranges that
the indicators show a significant
change in color.
• For test tubes B, C and D, using the
Henderson-Hasselbach equation, it
can be inferred that as the ratio of
the molarity OAc- (from NaOAc) to
that of HOAc increases, the pH also
increases thus making the solution
less acidic.

• pH = pKa + log [OAc-]

[HOAc]
• Because it has the highest ratio of OAc-
to HOAc, the solution in test tube D is
expected to be the least acidic while
the solution in test tube C as the most
acidic.
• Common ion effect can also account for
these. Because of the presence of the
common ion, OAc-, there will be a
suppression in the ionization of the acid
thus decreasing hydrogen ion
concentration and increasing the pH.
 Solution Observed
pH
A 0.01M HOAc 5
B 1mL 0.1M HOAc + 1mL 0.1M NaOAc + 4.8
8mL H20
C 1mL 0.1M HOAc + 0.1mL 0.1M NaOAc 3.2
+ 8.9mL H2O
D 0.1mL 0.1M HOAc + 1mL 0.1M NaOAc 6.8
+ 8.9mL H2O

• Therefore, our observations are correct!

 GUIDE QUESTIONS AND
ANSWERS
1.) Calculate the ionization constant of acetic acid using
colorimetric data.

pH of 0.01 M HOAc 5

pH = - log [H3O+]
5 = - log [H3O+]
[H3O+]= 10-5

[H3O+] of 0.01 M HOAc 1.00x10-5 M

[H3O+] of 0.01 M HOAc 1.00x10-5 M

To get Ka:
[H3O+] of 0.01 M HOAc 1.00x10-5 M

To get Ka:
HOAc → H+ + OAc-
Initial 0.01 0 0
Change - 1.00x10-5 + 1.00x10-5 + 1.00x10-5
Equilibrium 9.99x10-3 1.00x10-5 1.00x10-5
[H3O+] of 0.01 M HOAc 1.00x10-5 M

To get Ka:
HOAc → H+ + OAc-
Initial 0.01 0 0
Change - 1.00x10-5 + 1.00x10-5 + 1.00x10-5
Equilibrium 9.99x10-3 1.00x10-5 1.00x10-5

Ka = [H+][OAc-] = (1.00x10-5 )( 1.00x10-5 )= 1.00x10-8

[HOAc] 9.99x10-3
2.) Calculate the pH of three mixtures of HOAc and NaOAc
(solutions A, B and C) using the Henderson-
Hasselbalch equation and compare with the observed
pH. (Use the value of ionization constant of HOAc at
25°C.) Support your answers with computations.

B 1 mL 0.1 M HOAc + 1 mL 0.1 M NaOAc

+ 8 mL H2O
C 1 mL 0.1 M HOAc + 0.1 mL 0.1 M NaOAc
+ 8.9 mL H2O
D 0.1 mL 0.1 M HOAc + 1 mL 0.1 M NaOAc
+ 8.9 mL H2O
Ka of HOAc at 25°C = 1.8x10-5
pKa = -log Ka = - log (1.8x10-5) = 4.74

pH = pKa + log [conjugate base]

[acid]

The acid is HOAc and its conjugate base is OAc-.

For solution B:
1 mL 0.1 M HOAc + 1 mL 0.1 M NaOAc
+ 8 mL H2O
MA1 VA1 = MA2 VA2
(0.1M) (1mL) = [HOAc] (10mL)
[HOAc] = 0.01M

[NaOAc] = [OAc-]
MB1 VB1 = MB2 VB2
(0.1M) (1mL) = [OAc-](10mL)
[OAc-] = 0.01M
For solution B:
1 mL 0.1 M HOAc + 1 mL 0.1 M NaOAc
+ 8 mL H2O

pH = pKa + log [conjugate base]

[acid]
= 4.74 + log 0.01 M
0.01 M
= 4.74 + 0
pH = 4.74
For solution C:
1 mL 0.1 M HOAc + 0.1 mL 0.1 M
NaOAc + 8.9 mL H2O
MA1 VA1 = MA2 VA2
(0.1M) (1mL) = [HOAc] (10mL)
[HOAc] = 0.01M

[NaOAc] = [OAc-]
MB1 VB1 = MB2 VB2
(0.1M) (0.1mL) = [OAc-](10mL)
[OAc-] = 0.001M
For solution C:
1 mL 0.1 M HOAc + 0.1 mL 0.1 M
NaOAc + 8.9 mL H2O

pH = pKa + log [conjugate base]

[acid]
= 4.74 + log 0.001 M
0.01 M
= 4.74 + (-1)
pH = 3.74
For solution D:
0.1 mL 0.1 M HOAc + 1 mL 0.1 M
NaOAc + 8.9 mL H2O
MA1 VA1 = MA2 VA2
(0.1M) (0.1mL) = [HOAc] (10mL)
[HOAc] = 0.001M

[NaOAc] = [OAc-]
MB1 VB1 = MB2 VB2
(0.1M) (1mL) = [OAc-](10mL)
[OAc-] = 0.01M
For solution D:
0.1 mL 0.1 M HOAc + 1 mL 0.1 M
NaOAc + 8.9 mL H2O

pH = pKa + log [conjugate base]

[acid]
= 4.74 + log 0.01 M
0.001 M
= 4.74 + 1
pH = 5.74
 pH
Solution
observed calculated
0.01 M HOAc 5 ----
A 1 mL 0.1 M HOAc + 1 mL 0.1 M
NaOAc + 8 mL H2O (A) 4.8 4.74

B 1 mL 0.1 M HOAc + 0.1 mL

NaOAc + 8.9 mL H2O (B) 3.2 3.74

C 0.1 mL 0.1 M HOAc + 1 mL 0.1

6.8 5.74
M NaOAc + 8.9 mL H2O (C)

D
Conclusion
• The pH of a solution can be
approximated with the use of
colorimetry and pH indicators. But it
is also important to have to have
proper knowledge on which
indicator to be used on certain pH
ranges and their color transitions for
a successful colorimetric analysis.
Recommendation
• It is strongly advised to have
accurate measurements for the
preparation of buffer solutions to
have an efficient standard and also
for the solutions that will be used for
colorimetric analysis.
References
• Lemay, H., Brown, T., Bursten, B., & Burdge, J. (2004).
Chemistry: The Central Science. New Jersey: Pearson
Education South Asia Pte Ltd.
•  http://www.answers.com/topic/colorimetry#ixzz1MK1Yv3Sl
• http://www.ph-meter.info/pH-measurements-indicators
• http://www.chem.purdue.edu/gchelp/howtosolveit/equilibriu
m/buffers.htm
• http://www.biochemlab.cn/shiji/peizhi/20993.html
• http://medical-dictionary.thefreedictionary.com/Henderson-
Hasselbalch+equation
• http://www.inc.bme.hu/en/subjects/genchem/phdet2.pdf