CHY 47.

1 Basic Biochemistry, Laboratory

Activity 3
Buffer solutions

Activity 3A
Buffer Solutions
Individual Worksheet

Name: MACALAM, Glydel Jean V.

Date: October 10, 2022

Section Code: BSN 1C

Schedule: Tuesday & Thursday 10:00 AM – 1:00 PM

This part provides exercises for theoretical calculations. An online version of Figure 1. Worksheet
online version
this data sheet can be obtained by scanning the QR code in Figure 8 or through the
URL: https://forms.gle/z325BPiBdLrYf2Wo8.

Exercises
1. Can the following acid-conjugate base pairs be used as a buffer? Check the box corresponding to your
answer.
a. Ammonia and ammonium chloride ☒ Yes ☐ No
b. Acetic acid and potassium acetate ☒ Yes ☐ No
c. Nitric acid and sodium nitrate ☐ Yes ☒ No
d. Phosphoric acid and monopotassium phosphate ☒ Yes ☐ No
e. Chlorous acid and tris ☐ Yes ☒ No

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CHY 47.1 Basic Biochemistry, Laboratory

2. Consider a glycolic acid (HOCH2 COOH) and glycolate (HOCH2 COO− ) buffer system. Write the chemical
equation representing

a. The glycolic acid-glycolate equilibrium

b. The buffer system titrated (added) with a strong base

c. The buffer system titrated (added) with a strong acid

3. Prepare a 250-mL buffer with pH = 4.40 from solid reagents benzoic acid (C6 H5 COOH; M = 122.123
g mol−1 ) and sodium benzoate (C6 H5 COONa; M = 144.105 g mol−1 ). The p𝐾𝑎 for potassium dihydro-
gen phosphate is 4.202.

a. Write the balanced equation showing the equilibrium of the buffer solution.

b. Calculate the concentration ratio of benzoate to benzoic acid using the Henderson-Hasselbalch
Equation.

c. Determine the amounts (in mol) of C6 H5 COOH and C6 H5 COONa required to prepare the buffer.

d. Determine the mass of C6 H5 COOH and C6 H5 COONa required to prepare the buffer.

e. How is the buffer solution prepared? Write a step-by-step procedure.

- Dissolve 1 g of C6H5COOH
Dissolve 1.86g C6H5COO
Dilute into 250 mL Volumetric Flask

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CHY 47.1 Basic Biochemistry, Laboratory

4. A 100-mL buffer solution with pH of 4.80 is prepared as a stock solution. Using this stock buffer solu-
tion, three buffers are prepared by diluting a given volume of the stock buffer (as shown below) to
100 mL with water.
Volume used from
Buffer Final volume, mL
original solution, mL
1 5 100
2 10 100
3 20 100

a. Do they have the same pH? Check the box corresponding your answer.
☒ Yes ☐ No

b. Which is true about their absolute concentrations? Check the box corresponding your answer.
☐ [Buffer 1] > [Buffer 2] > [Buffer 3]
☒ [Buffer 3] > [Buffer 2] > [Buffer 1]
☐ [Buffer 3] = [Buffer 2] = [Buffer 1]
☐ [Buffer 3] = [Buffer 2] > [Buffer 1]
☐ [Buffer 3] > [Buffer 2] = [Buffer 1]

c. Which is true about their concentration ratios? Check the box corresponding your answer.
☐ [Buffer 1] > [Buffer 2] > [Buffer 3]
☐ [Buffer 3] > [Buffer 2] > [Buffer 1]
☒ [Buffer 3] = [Buffer 2] = [Buffer 1]
☐ [Buffer 3] = [Buffer 2] > [Buffer 1]
☐ [Buffer 3] > [Buffer 2] = [Buffer 1]

d. Which of the following is true about the ability of the three buffers to resist pH change when 1 mL
of HCl is added to each buffer? Check the box corresponding your answer.
☒ Ability to resist pH change: Buffer 1 < Buffer 2 < Buffer 3
☐ Ability to resist pH change: Buffer 1 > Buffer 2 > Buffer 3
☐ Ability to resist pH change: Buffer 1 = Buffer 2 = Buffer 3
☐ Ability to resist pH change: Buffer 1 < Buffer 2 = Buffer 3
☐ Ability to resist pH change: Buffer 1 = Buffer 2 < Buffer 3

e. Which solution has the highest buffer capacity? Check the box corresponding your answer.
☐ Buffer 1
☐ Buffer 2
☒ Buffer 3
☐ Their buffer capacities are equal

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CHY 47.1 Basic Biochemistry, Laboratory

5. From the stock solutions of 1.00-mol L−1 acetic acid and 1.00-mol L−1 of sodium acetate, six solutions
were prepared with the acetic acid and acetate concentrations shown in the table below. The p𝐾𝑎 of
acetic acid is 4.756.

-1 -1
Solution [Acetic acid], mol L [Acetate], mol L Final volume, mL
A 0.0000351 0 100
B 0.10 0.10 100
C 0.010 0.010 100
D 0.10 0.15 100
E 0.15 0.10 100

a. Determine the volume of the stock solutions of 1.00-mol L−1 acetic acid and 1.00-mol L−1 of so-
dium acetate needed to prepare the solutions. Write your answer in the table below.

[Acetic acid], Vol. acetic acid, [Acetate], mol Vol. of acetate, Final volume,
Solution -1 -1
mol L mL L mL mL
A 0.0000351 0.0351 0 0 100
B 0.10 10 0.10 10 100
C 0.010 1 0.010 1 100
D 0.10 10 0.15 15 100
E 0.15 15 0.10 10 100

b. Calculate for the initial pH of the solutions and write the pH in the table below.

-1 -1
Solution [Acetic acid], mol L [Acetate], mol L Initial pH
A* 0.0000351 0 4.756
B 0.10 0.10 4.756
C 0.010 0.010 4.756
D 0.10 0.15 4.93
E 0.15 0.10 4.58

* Use quadratic formula:

https://www.calculatorsoup.com/calculators/algebra/quadratic-formula-calculator.php

ax2 + bx + c = 0

c. Which solution(s) have the same pH?

- Solution A, B and C

d. Which solution is closest to the p𝐾𝑎 ?

- Solution A, B and C

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CHY 47.1 Basic Biochemistry, Laboratory

e. Calculate the pH of the solutions and the change in pH after adding 5.00 mL of 0.10-mol L−1 NaOH.

Solution Initial pH Final pH pH change

A 4.76 11.68 5.92
B 4.76 4.80 0.04
C 4.76 5.23 0.47
D 4.93 4.97 0.21
E 4.58 4.62 0.04

f. Calculate the pH of the solutions and the change in pH after adding 5.00 mL of 0.10-mol L−1 HCl.

Solution Initial pH Final pH pH change

A 4.76 2.32 - 2.44
B 4.76 4.71 -0.05
C 4.76 4.29 -0.47
D 4.93 4.90 -0.03
E 4.58 4.54 -0.04

g. Which solution is most resistant to pH change?

- Solution D

h. Which solution is least resistant to pH change?

- Solution A