DEPARTMENT OF BIOLOGY

FACULTY OF SCIENCE AND MATHEMATICS

SULTAN IDRIS EDUCATION UNIVERSITY

SBL 1023

TECHNIQUE IN BIOLOGY AND BIOCHEMISTRY LABORATORY

EXPERIMENT 2 : TITRATION

BY

BIL NAME STUDENT ID LECTURER

GROUP

1. NOR AIYNA SYERA BT MOHD ASRI E20161015981 B

Title :

Experiment 2 : Titration

Objective :

- To measure the pH and to standardise the acid solution

- To determine the difference in Ka of monoprotic and polyprotic acid

Equipment and materials

- Phenolphthalein indicator
- Buret
- Pipette
- 250ml beakers
- pH meter
- White paper towel

Methodology :

A) Titration of monoprotic acid ( acetic acid ) with NaOH

1. The buret was filled with 0.1 M NaOH. 25.00 ml of 0.1 M CH3COOH was pipet
into a 250 ml beaker and 4 drops of phenolphthalein indicator was added. The
beaker was placed on a white paper towel to best observe colour changes.
2. Titrate the solution by adding the NaOH titrant in 1-2 ml increments. The beaker
was swirl carefully with each addition.
3. The coloured form of the phenolphthalein will begin to stay for a while and then
disappear. At this point add the NaOH dropwise until the acetic acid is very light
colour. This is the endpoint for phenolphthalein.
4. The pH of the solution in the beaker at this end point was measured and recorded.
The pH probe was rinsed with distilled water and the probe tip was replaced into
its vial.
5. Any colour change observed during the titration was recorded. The pH and added
NaOH volume at that indicator’s end point should be used to estimate the target
point when conducting the following procedure.
 6. The pH/volume data was transferred to an excel file for later analysis. Store the
volumes in the A coloumn and the pH values in the B column of the spreadsheet,
beginning in cells A1 and B1.
7. Using the saved data in the excel file, prepare a plot of pH vs volume of NaOh
added to observe the equivalence point and half equivalence point.
8. To verify the equivalence point, the inflection point was determine by calculating
the change in Ph per change in volume, delta pH /delta V, for each recorded
volume, as shown in figure 2 for the NaOH/HCL titration. Plot delta pH/ delta V
vs the volume of the titrant added. Tp perform this, check that your volume and
pH values are in columns A and B on your excel spreadsheet. In cell C1, type:
=(A1 + A2)/2. Click on cell C1 and drag down to the C cell that is on the same
row as the last filled cell in columns A and B. column C now contains the average
volume of titrant between two readings.
9. In cell; D1, type: = (B2-B1)/ (A2-A1) which will calculate the delta pH/ delta v
values. Highlight cell D1 and drag to fill the remaining cells as you did with
column C. Column D now contains the values of delta pH/ delta v for each
volume reading. The inflection point is found by plotting delta pH/ delta v vs the
volumes in column C.
10. Once the volume at the equivalence point is known, the volume of the half
equivalence point can be found. Using the titration curve, the pH at this volume
can be determined. Thus pKa and hence for acetic acid can be calculated.

B) Titration of polyprotic acid (phosphoric acid ) with NaOH.

- Repeat all the steps above with phosphoric acid.
Result :

1. Part A ( acetic acid )

pH Volume of NaOH ( mL)

4.00 2.00
4.02 4.00
4.25 6.00
4.46 8.00
4.57 10.00
4.72 12.00
4.83 14.00
5.00 16.00
5.24 18.00
5.60 20.00
6.36 22.00
11.13 24.00
11.57 26.00
12.32 28.00
12.98 30.00
13.68 32.00
Table 6.1: Result for acetic acid vs NaOH titration
 Graph pH against volume of NaOH used

ml of titrant
35

30

25

20
pH

15
ml of titrant
10

0
0 2 4 6 8 10 12 14 16
volume of titrant

Graph 6.1

2. Part B ( phosphoric acid )

Volume of NaOH ( mL ) pH
0.00 1.30
2.00 1.97
4.00 2.00
6.00 2.03
8.00 2.15
10.00 2.20
12.00 2.24
14.00 2.27
16.00 2.34
18.00 2.42
20.00 2.50
22.00 2.58
24.00 2.68
26.00 2.78
28.00 2.89
30.00 3.01
32.00 3.16
34.00 3.35
36.00 3.69
38.00 4.61
40.00 5.87
42.00 6.25
44.00 6.48
46.00 6.63
48.00 6.73
50.00 6.84
52.00 6.87
54.00 7.00
56.00 7.10
58.00 7.18
60.00 7.29
62.00 7.39
64.00 7.49
66.00 7.59
68.00 7.70
70.00 7.84
72.00 8.01
74.00 8.22
76.00 8.66
78.00 9.20
80.00 9.93
82.00 10.72
84.00 11.04
86.00 11.27
88.00 11.44
90.00 11.55
Table 6.2: Result of phosphoric acid vs NaOH
 Graph pH against the volume of NaOH added

Titration of phosphoric acid

14

12

10

8
pH

6
pH
4

0
0 20 40 60 80 100
Volume of titrant (mL)

Graph 6.2

Calculation:

A) Titration of monoprotic acid (acetic acid) with NaOH

Moles of titrant = volume of titrant (L) X concentration of titrant (M)

= 0.023 L X 0.1 M

= =2.3 X 10-3 mol NaOH

Moles of titrant = moles of analyte

Moles of analyte = 2.3 X 10-3mol

Concentration of analytes = mol of analytes, mol / volumes of analytes, L

Concentration of analytes = 2.3 X 10-3mol / 0.025 L

= 0.092 mol/L
 pKa = - log Ka
pH = pKa

pKa = 4.80

Ka = 10 (-4.80 )

= 1.58 X 10-5

B) Titration of polyprotic acid (phosphoric acid) with NaOH

Moles of titrant = volume of titrant (L) X concentration of titrant (M)

= 0.038 L X 0.1 M
= 0.0038 mol NaOH

Moles of titrant = moles of analyte

Moles of analyte = 0.0038 mol

Concentration of analytes = mol of analytes, mol / volumes of analytes, L

Concentration of analytes = 0.0038 mol / 0.025 L

= 0.152 mol/L

pKa = - log Ka
pH = pKa

pKa = 2.27
Ka = 10 (-2.27)
= 5.37 X 10-3
Discussion

Titration is a common technique that used to quantitatively determine the concentration

of an identified analyte. We used phenolphthalein indicator for this experiment of titration.
Phenolphthalein is slightly soluble in water and usually dissolves in alcohol. From the
titration experiments, colour change from colourless to light pink. In this experiment, the
sample that we used is acetic acid and phosphoric acid.

For the titration of acetic acid is shown in Graph 6.1. Acetic acid which is a weak acid
has been titrated with a 0.1 M solution of NaOH solution. All differences in this experiment
between phosphorus and acetic acid are not affected by the type of the acid because both of
that acid is weak acid. It only affected by their category, whether they are monoprotic or
polyprotic.

In the acetic acid, the NaOH volume used is less than in phosphorus acid. Acetic acid is
a monoprotic acid since it has only one pKa point. By analysing graph of acetic acid, at the
infection point, the pH becomes 4.80 as 12.40 mL NaOH has been added. The titration of
phosphoric acid is shown in Graph 6.2. Phosphoric acid is a polyprotic acid. When some
NaOH is added, the solution now contains acid and conjugate bases (dihydrogen phosphate).
This conjugate base act as a buffer which resists the change in pH. Phosphorus acid which is
can release three protons per molecule need more volume of NaOH than the acetic acid.

From this experiment, the result was not too accurate compare to the theoretical value.
This might be happened because of the pH meter that we used is not measured the accurate
pH reading. There are several precaution steps to make sure the data collected is correct and
it also to ensure the safety. We must to be ensuring there is no bubble trapped at the tip of the
burette during fill the sodium hydroxide solution and we need measure the reading sharply to
avoid the error. Calibrate the pH meter before using it and make sure the beaker is swirled
regularly during the experiment.

Conclusion

From this experiment, we can know the differences between the monoprotic and polyprotic
acids. We also know how to calibrate the pH using pH meter.