BUFFER ACTIVITY

The following hypothetical laboratory experiment will help you concretely visualize how
buffers work. Consequently, this will also help you to apply the Henderson-Hasselbalch
equation in predicting the resulting pH values.

You were tasked to prepare an acetic acid-acetate buffer in twelve (12) test tubes. The
concentrations of the components are as follows:

Test
[salt] [acid] Log ratio pH
tube
1 2 × 10-4 8 × 10-4
2 2.5 × 10-4 7.5 × 10-4
3 3 × 10-4 7 × 10-4
4 3.5 × 10-4 6.5 × 10-4
5 4 × 10-4 6 × 10-4
6 4.5 × 10-4 5.5 × 10-4
7 5.5 × 10-4 4.5 × 10-4
8 6 × 10-4 4 × 10-4
9 6.5 × 10-4 3.5 × 10-4
10 7 × 10-4 3 × 10-4
11 7.5 × 10-4 2.5 × 10-4
12 8 × 10-4 2 × 10-4

Calculate the log ratio and pH using the Henderson-Hasselbalch equation. Plot the log
ratio (x-axis) against the pH (y-axis).

If you added a 1 × 10-4 M NaOH to the buffer solution in test tube 5, what will be the
resulting pH? Did the solution resist a change in pH?

If you added a 1.5 × 10-5 M HCl to the buffer solution in test tube 11, what will be the
resulting pH? Did the solution resist a change in pH?
QUESTIONS
1. What are the three most important buffers in the human body? Briefly describe each of their
function.
2. Explain how the molar concentration of H2CO3 in the blood would change if the partial
pressure of CO2 in the lungs were to increase?
3. Explain how the molar concentration of H2CO3 in the blood would change if the partial
pressure of CO2 in the lungs were to decrease?
4. Explain how the molar concentration of hydronium ion in the blood would change under each
of the conditions in Questions 2 and 3.
5. Answer the following:
a. Calculate the pH of a mixture of 0.25 M acetic acid and 0.1 M sodium acetate.
b. What happens to the pH in the preceding problem if the mixture consists of 0.1 M acetic
acid and 0.25 M sodium acetate?
c. Calculate the ratio of lactic acid and lactate required in a buffer system of pH 5. The pKa of
lactic acid is 3.86.
d. During the fermentation of wine, a buffer system consisting of tartaric acid and potassium
hydrogen tartrate is created as the result of a biochemical reaction. Assuming that at some
point in the process, the concentrations of the two components are approximately equal,
determine the pH of the wine. The pKa of tartaric acid is 2.96.