Department of Pure and Applied Chemistry

College of Arts and Sciences

Visayas State University
Visca, Baybay City, Leyte

Name: Ana Luisa C. Laurente Date Performed: June 11, 2019

Lab Schedule: 7:00 – 10:00 (M-F) Date Submitted: July 15, 2019

Group No: 6 Score:

Experiment No. 2

pH and Buffer System

A. Abstract

We made a buffer solution by mixing acetic acid and sodium acetate and got
the desired pH amount in our assigned pH. We made a buffer solution using graduated
cylinders and pipette to measure the volumes of solutions to mix and it produced
buffers with significant buffer solution. By conducting the experiment, our results reveal
that a buffer solution always contains significant concentrations of a weak acid and a
conjugate base, and that a buffer solution changes in PH when a strong acid or base
is added to it. Buffers can be prepared by mixing a weak acid and the salt. For our
experiment, we observed that the ph changes when 2ML of 1M of NaOH were added.

B. Introduction

The main purpose of this experiment was to be able to accurately prepare

different pH buffer solutions and identify which has the higher buffer capacity and to
measure the pH accurately utilizing the calibrated pH meter. Calculation of the ratio of
conjugate base to weak acid. utilizing the Henderson-Hasselbalch was performed as
well as the titration of the assigned amino acid.

pH also known as potential Hydrogen is a measure of the hydrogen ion

concentration of a solution. Solutions with a high concentration of hydrogen ions have
a low pH and solutions with a low concentrations of H+ ions have a high pH. A pH
value can range from 1 to 14 with the pH value 7 as the neutral (middle) point. Values
that are below 7 is an indication for acidity which increases as the pH value decreases.
While values that belong above the pH of 7 indicates alkalinity or basic. Determining
the pH value is the most convenient way to identify whether a solution is basic or acidic.
A buffer is an aqueous solution that resists changes in pH upon the addition of a small
amount of acid or base. The action of a buffer is dependent on the common ion effect.
An example of a buffer is the blood which is present in all living organisms.

C. Methodology

1. Calibration of the pH meter

2. Factors affecting buffer capacity
 Effect of concentration of the buffer using 0.1 M phosphate buffer and
acetate buffer solution
 Record the pH of each buffer solution. Add 2 mL of NaOH to each of the
samples assigned to groups
  Effect of the ratio of the conjugate base to the weak acid from Henderson-
Hasselbach equation, calculate the ratio of dihydrogen phosphate and
monohydrogen phosphate and the ratio of acetic acid and acetate
required to produce a solution.
3. Choice and preparation of a buffer system.
 Choosing the proper buffer solution.
 Preparation of the chosen buffer system.
4. Titration of an amino acid
 Pipette in 10mL of assigned amino acid.
 Stir well and measure the pH after each addition.

D. RESULTS

Factors affecting Buffer Capacity

Case 1. Effect of Concentration of buffer (O.1 M Acetate Buffer)

Table 1. Prepared (Phosphate Buffer) and pH change upon addition of strong base
(NaOH)

pH
upon Amount
Expected Actual Difference Magnitude Equivalent
Required addition of buffer
pH pH in pH of pH % of pH
concentration of solution
NaOH
0.005 M 7.2 6.9 -0.3 7.7 +0.8 11.59% 12.5
0.05 M 7.2 7.1 -0.1 7.3 +0.2 2.81% 12.5
0.1 M 7.2 6.9 -0.3 7.0 +0.1 1.44% 25

Case 2. Effect of Concentration of Buffer

Table 2. Prepared (Acetate Buffer) and pH change upon addition of strong base
(NaOH)

pH upon Amount
Required Expected Actual Difference addition Magnitude Equivalent of
concentration pH pH in pH of of pH % of pH Buffer
NaOH solution
0.005 M 4.7 4.4 -0.3 6.8 +2.4 4.54% 12.5
0.05 M 4.7 4.4 -0.3 4.6 +0.2 4.54% 12.5
0.2 M 4.7 4.4 -0.3 5.6 +1.2 25% 25

Effect of the ratio of the conjugate base and weak acid

Case 1. Using potassium hydrogenphosphate and potassiumdihydrogenphosphate

treired

Table 1. Prepared (Phosphate Buffer) and pH change upon addition of strong base
(NaOH)
 Amount of Amount of pH with
Expected Magnitude Equivalent
Group K2HPO4 KH2PO4- 0.1 M
pH pH change pH change
(mL) (g) change
1 6.2 13 1.858 6.8 +0.6 8.82%
2 7.2 7.4 0.10 7.6 +0.4 5.26%
3 8.2 136.15 0.208 8.3 +0.1 1.20%
Case 1. Using potassium hydrogenphosphate and potassiumdihydrogenphosphate
treired

Table 1. Prepared (Acetate Buffer) and pH change upon addition of strong base
(NaOH)

Amount of Amount of pH with

Expected Magnitude Equivalent
Group K2HPO4 KH2PO4- 0.1 M
pH pH change pH change
(mL) (g) change
1 6.2 5 120 3.9 +0.2 5.13%
2 7.2 2.67 69.8 4.9 +0.2 4.08%
3 8.2 0.5 0.208 6.2 +0.6 8.07%

E. Titration of Amino Acid (Alanine)

Alanine 4.6 21 6.8 43

pH Amino 4.7 22 6.9 44
Acid 4.8 23 7 45
2.7 6 4.9 24 7.1 46
2.8 6.5 5 25 7.2 47
2.9 7 5.1 26 7.3 48
3 7.5 5.2 27 7.4 49
3.1 8 5.3 28 7.5 50
3.2 8.5 5.4 29 7.6 51
3.3 9 5.5 30 7.7 52
3.4 9.5 5.6 31 7.8 60
3.5 10 5.7 32 7.9 63
3.6 11 5.8 33 8 66
3.7 12 5.9 34 8.1 69
3.8 13 6 35 8.2 72
3.9 14 6.1 36 8.3 75
4 15 6.2 37 8.4 78
4.1 16 6.3 38 8.5 81
4.2 17 6.4 39 8.6 84
4.3 18 6.5 40 8.7 87
4.4 19 6.6 41 8.8 90
4.5 20 6.7 42 8.9 93
 9 96 9.7 139 10.4 209
9.1 99 9.8 149 10.5 219
9.2 104 9.9 159 10.6 229
9.3 109 10 169 10.7 239
9.4 114 10.1 179 10.8 249
9.5 119 10.2 189 10.9 259
9.6 129 10.3 199

DISCUSSION

Calibration of the pH meter

The calibration of the pH meter follows a procedure. The electrode must be placed into
a pH 7.00 buffer solution. When using the pH meter, the electrode must first be washed by
distilled water and dried gently by a tissue paper. There are two pH indicators with the colors
of red and green. Readings of the electrode must first be stabilized before recording them to
obtain accurate and precise data.

Factors Affecting Buffer Capacity

Buffer solutions have the useful property of resisting changes in pH when hydrogen
ions are added to or subtracted from the solution by chemical reactions or by other means.
This is because a buffer solution contains both an acidic and a basic component. Buffer
solutions are important because many reactions will work best within a certain range of pH
values.

In table 1 (acetate buffer), the results have quiet similar pH value. When the original
concentration of buffer is high the account for magnitude change in pH is lower. This is the
effect of concentration of buffer. Upon conducting,

A 25 mL 0.005 M acetate buffer was prepared and initial pH was recorded having pH of
4.4 using the calibrated pH meter. An addition of 2 mL of 0.1 M NaOH was done and pH was
recorded having pH of 6.8 with the magnitude change in pH accounted for as (+) 2.4. A
separate acetate buffer in 25 mL 0.05 M concentration was prepared and the same procedure
was done, from pH 4.4 the pH a rise of (+) 0.2 magnitude making it to achieve pH 4.6 after
adding 2 mL of 0.1 M NaOH. The same goes for the 25 mL 0.10 M acetate buffer which was
separately prepared and the same procedure was done. From pH value of 4.4, an increase of
(+) 1.2 magnitude after adding 2 mL 0.1 M NaOH was recorded making it pH 5.6. 0.05 M buffer
concentration as the least magnitude change in pH making it as the buffer with greatest
capacity to resist change in pH compared to the other two.

Titration of Amino Acid (Alanine)

Alanine is one of the most widely used for protein construction and is involved in the
metabolism of tryptophan and vitamin pyridoxine. Alanine was pipetted into a 50 mL beaker

The pH was adjusted to 1.5. 0.1M NaOH in 0.5 mL increments and was added until
the pH value of 12 was reached. The exact volume of each increment rise is recorded and the
results are graphed. However, some changes were done by changing the increments from 1
mL to 3 mL to 5 mL and lastly 10 mL Changes increments were done to maximize time, to
wait for pH 1.5 to rise to pH 12 with just 0.5 mL increments. Approximately 379 mL of 0.1 M
NaOH was used to achieve a pH of 12.

F. Conclusion

With help of the Henderson-Hasselbalch equation it concludes that the factors

affecting buffer capacity is the actual concentrations of acid and base and on the pKa and
the [acid]/[base] ratio is of importance because the greater the concentration of the weak
acid and its conjugate base, the greater the buffer capacity.

It is also concluded that the pH of buffer solutions depends on the amount of acid or
base added to the buffer solution. In determining ph levels, calibrated ph meter choosing
the appropriate buffer system and titration of amino acid is very important.

G. References

http://academic.brooklyn.cuny.edu/biology/bio4fv/page/ph_def.htm

http://www.businessdictionary.com/definition/pH-scale.html

http://www.savitapall.com/Acids-Bases/Notes/Buffers%20and%20pH%20of%20Buffers.pdf

https://pubchem.ncbi.nlm.nih.gov/compound/L-Alanine