Laboratory Report No.

1
pH and Buffers

Esconde, Yvonne Keithlene J. BS Nursing 1A

Grp. 3/ Thurs/ Aug. 16, 2018
Rating: ________

I. Objectives

1. To illustrate the buffering properties of phosphates and acetates.

2. To provide the students a sense of how buffers work.

II. Introduction

The term pH was introduced in 1909 by Sorensen, who defined pH as negative log of
hydrogen ion concentration. In first year chemistry, one has learned to calculate the pH
of solutions by (a) calculating hydrogen ion concentration, (b) calculating the base 10
log of hydrogen ion, and (c) pH is the negative of the value found in base 10 log. This
information will be adequate for most biochemical purposes of this biochemistry
experiment.
According to Bronsted-Lowry definition, acids are proton donors and bases are proton
acceptors. A distinction was made between strong acids, which completely dissociate
in solution, and weak acids, which does not completely dissociate in solution. A
similar distinction was likewise made between strong and weak bases. Many
biochemical are weak acids. It is then important to know how to calculate the pH of
acidic and basic solutions.

III. Materials

A. Equipment
• (4) 50-mL Beaker
• (2) 10-mL Volumetric Pipette
• (1) 10-mL Graduated Cylinder
• (2) 200-mL Beaker
• pH meter

B. Reagents
• 1M KH2PO4
• 1M CH3COOH
• 1M HCl
• 1M K2HPO4
• 1M CH3COO-Na+
IIII. Methodology (Schematic Diagram)
A.

B. Buffering Effect
V. Data and Results

Mixture pH Meter pH Paper

KH2PO4 / K2HPO4 6.8 7
CH3COOH / CH3COO-Na+ 5.7 4
Distilled Water (Beaker 1) 7.5 6
Distilled Water + HCl (Beaker 2) 4.3 4
Phosphate Buffer + HCl (Beaker 3) 6.5 7
Acetate Buffer + HCl (Beaker 4) 5.8 4

VI. Discussion

Buffers are special solutions that can maintain a certain range of hydrogen ion concentration
(its pH levels) for a substance upon the addition of acidic or basic components. These are
created by combining a weak acid or a weak base with its salt (Britannica, 2013). An
example of a common buffer is CH3COOH + CH3COO-Na-, or acetic acid and sodium
acetate, which was also used in the experiment.
In the experiment, HCl was added to all beakers except Beaker 1, which would serve as a
basis for the results of Beaker 2, wherein the pH level of the distilled water went from 7.5 to
4.3 when no buffer was also added. The Phosphate Buffer suffered less changes,
maintaining a range of 6-7 throughout the experiment. And the same thing was applied to
the Acetate Buffer, as it also maintained its initial pH level even after HCl was added to it.

VII. Conclusion

As one can observe in the results, there was only a slight difference in the pH levels of the
mixtures with buffers when hydrochloric acid was added, considering the fact that it is a
strong acid and could have done more intensive changes. On the other hand, the distilled
water without any buffers underwent a significant decrease in its pH levels. The buffers
were effective in their role to maintain the pH levels despite HCl’s own acidity levels.

VIII. Assessment (Q&A)

1. Show the equilibrium equations for the ionization of acetic acid and KH2PO4.

CH3COOH (aq) + H2PO4-(aq) <=> H3PO4 (aq) + CH3COO-(aq)

2. Derive Hendersen-Hasselback equation.

3. An acetate buffer was prepared by mixing 10 mL of 0.1 M acetic acid and 100 mL of 0.1 M
sodium acetate. What is the pH of the buffer solution?

(M1V1) = (M2V2)
(0.1M acetic acid) (0.01mL) = (?) (0.11mL)
?= 0.001/0.11 M = 0.009
CH3COOH (0.1M sodium acetate) (0.1mL) = (?) (0.11mL)
? = 0.01/0.11 M = 0.09
CH3COOHNa+ pH = pKa + log [salt]/ [acid] = 4.74 + log [0.09]/[0.009] = 5.744.
4. Can a buffer solution be prepared from a mixture of NaNo3 and HNO3? Explain.
No. In order to prepare a buffer, the salt must be mixed with a weak acid or base. NaNo3 and
HNO3, when combined, unfortunately forms a strong acid and salt mixture, and will thus not be
inductive in the creation of a buffer.

IX. References

Britannica. (2013). Buffer. Chemistry. Retrieved

from https://www.britannica.com/science/buffer-chemistry
Mehta, A. (2012). Hendersen-Hasselback equation: Derivation of pKa and
pKb. Pharmaxchange. Retrieved
from https://pharmaxchange.info/2012/10/henderson%E2%80%93hasselbalch-equation-
derivation-of-pka-and-pkb/

X. Certification / Conforme

Esconde, Yvonne Keithlene J.

Signature over printed name

August 23, 2018

Date Submitted: