EXPT 11.

pKa of pH Indicators

[Key Contents]
- acid dissociation constant, pKa
- buffer, Henderson-Hasselbalch equation
- spectrophotometry, Beer's law
- pH indicator
[References]
Principles of Modern Chemistry, 6th Ed. (Oxtoby et al.)
Ch 15. Acid-Base Equilibria
Chemistry for Life, Chemistry for Better Life (Kim et al.)
Ch 9. Equilibrium Reactions
[Goal]
- to understand basic principles of acid dissociation equilibrium
-

to

learn

to

use

Henderson-Hasselbalch

equation

in

acid

dissociation equilibrium
- to learn basic principles of spectrophotometry
[Background]
A typical pH indicator (HInd) is a weak acid that undergoes acid
dissociation in solution with an accompanying color change.
HInd Ind- + H+
The color of the solution is determined by the concentration ratio
between HInd and Ind-. The concentration of HInd and Ind- can be
measured using Beer's law.
A

= bC

: molar extinction coefficient,

b: path length,

A: absorbance

You will first obtain absorption spectrum of the indicator at acidic

and basic pH and select a wavelength where HInd absorbs strongly
and

Ind-

absorbs

little.

Then

you

will

measure

the

decreased

absorbtion at this wavelength as the pH of the indicator solution is

increased gradually using buffers of different pH. The decrease in
absorbance corresponds to the concentration of Ind-. The pKa of HInd
is the pH at which [HInd] and [Ind-] are the same. You can
determine pKa of the indicator by plotting log([Ind-]/[HInd]) against
pH.
[Apparatus and Chemicals]
spectrophotometer, cuvette, test tubes, 10 mL pipet, Pasteur pipet
0.01 mM bromophnol blue (BPB) solution, 0.02 mM phenolphthalein
solution
0.1 N HCl solution, 0.1 N NaOH solution, 10 mM buffer solutions (pH
3.4, 3.7, 4.0, 4.3, 4.6, 7.0, 8.5, 9.0, 9.5, 9.8)

[Procedure]
Expt 1. pKa of Bromophenol Blue

1) In separate test tubes, prepare following mixtures.

mixture 1 : 2.0 mL 0.05 mM BPB soln + 2.0 mL 0.1 N HCl soln
mixture 2 : 2.0 mL 0.05 mM BPB soln + 2.0 mL pH 3.4 buffer
mixture 3 : 2.0 mL 0.05 mM BPB soln + 2.0 mL pH 3.7 buffer
mixture 4 : 2.0 mL 0.05 mM BPB soln + 2.0 mL pH 4.0 buffer
mixture 5 : 2.0 mL 0.05 mM BPB soln + 2.0 mL pH 4.3 buffer
mixture 6 : 2.0 mL 0.05 mM BPB soln + 2.0 mL pH 4.6 buffer
mixture 7 : 2.0 mL 0.05 mM BPB soln + 2.0 mL pH 7.0 buffer
2) Using a spectrophotometer, obtain absorption spectrum from mixture
1 and 7. Read absorbance between 450-650 nm at 25 nm intervals. Read
absorbance at 5 nm intervals at the absorption peak. Put the solution
back to the test tube. Determine wavelength for maximum absorption (
max)

for HInd from mixture 1. Determine max for Ind- from mixture 7.

3) From mixture 2~6, read absorbance at both max.

4) Visually, estimate the pH where the color of the solution is the mixed
color of mixture 1 and 7 in equal volume.
5) Test of Beer's Law : In a test tube, mix 2 mL of mixture 1 with 2 mL
of 0.1 N HCl solution to make 0.025 mM BPB solution and read
absorption at both max. Next, mix 2 mL of the 0.025 mM BPB solution
with 2 mL of 0.1 N HCl solution to make 0.0125 mM BPB solution and
read absorption at both max. Finally, mix 2 mL of the 0.025 mM BPB
solution with 2 mL of the 0.0125 mM BPB solution to make 0.0188 mM
BPB solution and read absorption at both max.

HO

Expt 2. pKa of Phenolphthalein

OH

1) In separate test tubes, prepare following mixtures.

O

phenolphthalein

mixture 1 : 2.0 mL 0.02 mM phenolphthalein + 2.0 mL pH 7.0 buffer

mixture 2 : 2.0 mL 0.02 mM phenolphthalein + 2.0 mL pH 8.5 buffer
mixture 3 : 2.0 mL 0.02 mM phenolphthalein + 2.0 mL pH 9.0 buffer
mixture 4 : 2.0 mL 0.02 mM phenolphthalein + 2.0 mL pH 9.5 buffer
mixture 5 : 2.0 mL 0.02 mM phenolphthalein + 2.0 mL pH 9.8 buffer
mixture 6 : 2.0 mL 0.02 mM phenolphthalein + 2.0 mL 0.1 N NaOH
2) Using a spectrophotometer, obtain absorption spectrum from mixture
1 and 6. Read absorbance between 450-650 nm at 25 nm intervals. Read
absorbance at 5 nm intervals at the absorption peak. Put the solution
back to the test tube. Determine max for Ind- from mixture 6.
3) From mixture 1~5, read absorbance at max.
4) Visually, estimate the pH where the color of the solution is the mixed
color of mixture 1 and 6 in equal volume.
5) Test of Beer's Law : In a test tube, mix 1.5 mL of mixture 6 with 1.5
mL of 0.1 N NaOH solution to make 0.01 mM phenolphthalein solution

and read absorption at max. Next, mix 1.5 mL of the 0.01 mM

phenolphthalein solution with 1.5 mL of pH 7.0 buffer solution to make
0.005 mM phenolphthalein solution and read absorption at both max.
Finally, mix 1.5 mL of the 0.01 mM phenolphthalein solution with 1.5 mL
of

the

0.02

mM

phenolphthalein

solution

to

make

0.015

mM

phenolphthalein solution and read absorption at both max.

[Data Analysis]
Expt 1. pKa of Bromophenol Blue

1) Draw an absorption spectrum for mixture 1 and determine max and

at max. Calculate [HInd] from absorbance at max.
2) Plot [HInd] and [Ind-] againt the molar concentration of BPB. Is
the Beer's law valid?
3) For mixture 2~6, calculate

[HInd]

at

different

pH's

from

absorbance at max.
4) Calculate [Ind-] as difference between [HInd] for mixture 1 and
[HInd] at different pH's.
5) Plot log[Ind-]/[HInd] againt pH and determine pKa of BPB as the pH
where log[Ind-]/[HInd] is zero.
6) Draw an absorption spectrum for mixture 7 and determine max and
at max. Calculate [Ind-] from absorbance at max.
7) For mixture 2~6, calculate

[Ind-]

at

different

pH's

from

absorbance at max.
8) Calculate [HInd] as difference between [Ind-] for mixture 7 and

[Ind-] at different pH's.

9) Plot log[Ind-]/[HInd] againt pH and determine pKa of BPB as the pH
where log[Ind-]/[HInd] is zero.
10) Compare pKa values obtained by both methods. How good is the
agreement? Compare the average of the two pKa values with the
literature value of Ka, 1.5 x 10-4.

Expt 2. pKa of Phenolphthalein

1) Draw an absorption spectrum for mixture 6 and determine max and

at max. Calculate [Ind-] from absorbance at max.
2) Plot [Ind-] againt the molar concentration of BPB. Is the Beer's
law valid?
3)

Plot

log[Ind-]/[HInd]

againt

pH

and

determine

pKa

of

phenolphthalein as the pH where log[Ind-]/[HInd] is zero.

4) Compare pKa values obtained with the literature value of 9.3. Discuss
why phenolphthalein is a good indicator for titrating strong acid with
strong base.

[Additional Material]
Phenol Red and Phenolphthalein

pH 5.5

8.0

phenol red

phenolphthalein

10.0

12