DEPARTMENT OF PURE AND APPLIED CHEMISTRY

VSU, Visca, Baybay City, Leyte

Name: Essielve Batistil Date Performed: 3-15-23

Offering No. and Lab Schedule: R083 (W 10:00–1:00) Date Submitted: 3-24-

23

Group No: 1

Exercise No 1: pH and Buffer Systems Review

Objectives:

 To calibrate a pH-measuring device, multi-parameter meter, by three-point

calibration.

 To apply the concept behind the Henderson-Hasselbalch equation in choosing an

appropriate buffer as well as its preparation.

 To titrate an amino acid and discuss its relation to the concept of buffer systems.
 RESULTS AND DISCUSSION

A. Calibration of pH meter

Three-point calibration was done in pH buffer solutions following the 7, 4, and 10

calibrating sequence. Calibration at different values along the pH scale sets the pH meter to

its standard setting thus ensuring the accuracy of the resultant data upon experimental

measurement.

B. Preparation of a Buffer solution

Table 1. Preparation of 0.1 M Phosphate buffer (HPO42-/ H2 PO4-)

Buffer molarity Buffer’s volume Mass of salt Mass of acid Measured pH of

(K2HPO4) (KH2 PO4) the buffer

0.1 M 25 mL 49.9 g 13.6 g 7.53

The buffer solution was prepared from the salt form of dihydrogen phosphate

(H2PO4-) and hydrogen phosphate (HPO42-), both of which represented the conjugate base and

weak acid respectively. Choosing the appropriate buffer system stems from the proximity of

the required pH to the pKa of its weak acid such that, pH must fall within the range equal to

pKa ± 1. Since the pKa of dihydrogen phosphate is 7.20, it is an appropriate component for a

buffer system with pH around 7. The ratio of the components were determined by using the
Henderson-Hasselbalch equation and the calculated ratio served as the basis for the amount

of the components to be put into the system.

Calculation for the preparation of Buffer:

pH = pKa+log ¿ ¿

pH = pKa+log ¿ ¿

7.2=6.86 +log ¿ ¿

Mass of K2HPO4:

2.1877 mol g 1
× 228.23 × =49.9 g
L mol 1000 mL

Mass of KH2 PO4:

1mol 136.09 g 1
× × × 100 mL=13.6 g
L mol 1000 mL

C. Titration of Amino acid

Table 2. Data for the pH of amino acid upon its titration against 0.1 M HCl
 0.1 M HCl (mL) pH 0.1 M HCl (mL) pH
0 6.57 10.5 3.68
0.3 6.4 11.5 3.65
0.6 5.26 12.5 3.65
0.9 4.92 13.5 3.62
1.2 4.66 15.5 3.61
1.5 4.52 17.5 3.58
1.8 4.43 19.5 3.54
2.1 4.36 21.5 3.55
2.4 4.29 23.5 3.5
2.7 4.24 25.5 3.5
3 4.18 28.5 3.5
3.3 4.14 32 3.44
3.6 4.09 35.5 3.43
3.9 4.04 39 3.4
4.2 4.02 43 3.37
4.5 4 47 3.34
5 3.96 50 3.31
5.5 3.91 55 3.29
6 3.87 60 3.36
6.5 3.84
7 3.81
7.5 3.79
8.5 3.76
9.5 3.73

Table 3. Data for the pH of amino acid upon its titration against 0.1 M NaOH
std. 0.1 M NaOH (mL) pH std. 0.1 M NaOH (mL) pH
0 6.82 10.3 11.81
0.3 7.84 10.8 11.89
0.6 8.28 11.3 11.77
0.9 8.56 11.8 11.86
1.2 8.88 12.3 11.9
1.5 9.11 12.8 11.96
1.8 9.19 13.3 12
2.1 9.28 13.8 12.03
2.4 9.45 14.3 12.08
2.7 9.55 14.8 12.11
3 9.68 15.3 12.14
3.3 9.78 15.8 12.18
3.6 9.92 16.3 12.19
3.9 10.02 17.3 12.22
4.2 10.12 18.3 12.24
4.5 10.17 19.3 12.27
5 10.38 20.8 12.32
5.8 10.64 21.8 12.29
6.3 10.92 22.8 12.32
6.8 11.03 23.8 12.37
7.3 11.2 24.8 12.41
7.8 11.31 25.8 12.44
8.3 11.44 26.8 12.47
8.8 11.54 27.8 12.5
9.3 11.67
9.8 11.76
 14

12

10

8
pH

0
0 5 10 15 20 25 30
0.1 M NaOH (mL)

Figure 1. Titration curve of lysine with a base

4
pH

0
0 5 10 15 20 25

0.1 M HCl (mL)

Figure 2. Titration curve of lysine with an acid

The data for the pH of the amino acid, lysine, against the amount of acid or base

added was tabulated above. An amino acid is known to behave as a buffer system and its

capability to act as a buffer attributes to its amphoteric nature by having both a carboxylic
 and amino group. Whichever titrant was added, either base or acid, the amino acid can

neutralize the effect of the titrant.

The amino acid being subjected had most of its components already in zwitterion

forms for it initially has a pH of around 6. Unless the amino acid is initially at its extreme pH,

it cannot provide the curve with a single titrant alone hence, in the experiment, two titrants,

HCl and NaOH were utilized so as to look for the two sides of the titration curve as presented

in Figure 1 and Figure 2. To create the usual form of the titration curve of a buffer system,

both of the curves were combined as presented in Figure 3. It was noted however that the

negative sign will be disregarded for its value represents the volume of acid added.

13
12
11
10
9
8
7
pH

6
5
4
3
2
1
0
-60 -40 -20 0 20 40 60

0.1 M acid and base added

pH of Lysine against HCl pH of Lysine against NaOH

Figure 3. Combined titration curves of Lysine

The pKa is the point at which equivalence point is reached. For instance, when a basic

titrant is added to amino acid at its extreme low pH, the carboxylic group (-COOH) becomes

deprotonized until such a time when the functional group (-COOH) and its ionized form (-
COO-) form are equal. Eventually, the second pKa is due to the equivalence of the amino

group (–NH2) and its ionized form (–NH3+ ).

According to literature, the pKa1 and pKa2 of lysine is 2.16 and 9.06. However, it was

not clearly manifested in the titration curve. This inconsistency to the supposed trend of the

titration curve must have stemmed from experimental errors and faulty techniques.

CONCLUSION

The experiment was able to meet the objectives for it had involved the calibration of

the pH device, preparation of a buffer and titration of amino acid. Errors of conduct were

however incurred.

References

Garcia, R.G. (2020). pH and Buffer System. Lab Report.

https://www.academia.edu/88569707/pH_and_BUFFER_SYSTEM

Amino Acid. n.d. Accessed 24 March 20223 from

https://fac.ksu.edu.sa/sites/default/files/amino_acid_titration.pdf