SBL1023

TECHNIQUES IN BIOLOGICAL AND

BIOCHEMISTRY LABORATORY

TITRATION

NAME: NUR IZZATIE BINTI KHAIZAAR

MATRICS NUMBER: E20161013729
GROUP: A
LECTURERS NAME: DR. SHAKINAZ BINTI DESA
 TITRATION
INTRODUCTION
Titration or also known as titrimetry is about laboratory method of quantitative chemical
analysis that is used to determine the unknown concentration of an identified analyte.
In this experiment, equivalence point is very important information about acid
dissociation constant, Ka, of the acid being titrated. The colour change occurs when
the end point reach as we added the proper indicator in the analyte. The pH meter is
used to record the value of pH as the titrant is added. The recorded data could be
plotted into graph between pH values against volume of titrant added to the analyte
and form a titration curve. From the titration curve (s-shape), we could find the
equivalence point.

OBJECTIVE
To differentiate the monoprotic acid and polyprotic acid by using titration method.

METHODOLOGY
A) Titration of monoprotic acid (acetic acid) with NaOH
1. The burette is filled with 0.1M NaOH. 25.00 mL 0.1M CH3COOH is pipetted
into a 250 mL beaker and 3-4 drops of phenolphthalein indicator are added.
The beaker is placed on a white paper towel to best observe colour
changes.
2. The solution is titrated by adding the NaOH titrant in 2 mL increments. The
beaker is swirled carefully with addition.
3. The coloured of the phenolphthalein is formed when it begin to stay for a
while and then disappear. At this point, the NaOH is added dropwise until
the acetic acid is a very light colour. This is the endpoint for phenolphthalein.
4. Then, the pH probe is rinsed with distilled water and the probe tip is replaced
into vial.
5. Any colour change observed is recorded during the titration. The pH and
added NaOH volume at that indicators endpoint is estimated the target point
when conducting the following procedure.
6. The pH and volume is transferred into Excel file for later analysis. The
volume is stored in A column and the pH values in the B column of the
spreadsheet, beginning in cells A1 and B1.
7. The saved data is used in the Excel file to prepare a plot of pH vs. volume
of NaOH added in order to observe the equivalence point and half
equivalence point.
8. Then, the graph plotted is used to find concentration of analyte, pKa and Ka.
9. All the steps are repeated by using phosphoric acid.
RESULTS
Acetic acid
Volume of titrant added, mL pH

2.0 3.18
4.0 3.67
6.0 3.90
8.0 4.09
10.0 4.37
12.0 4.53
14.0 4.73
16.0 4.90
18.0 5.08
20.0 5.38
22.0 6.06
24.0 11.27
26.0 11.70
28.0 11.90
30.0 12.04

Graph of pH value versus volume titrant of added

14

12

10

8
pH value

6 pH

0
0 5 10 15 20 25 30 35
volume of titrant added, mL
Equivalence point
(11.2 pH + 6.0 pH) 2 = 8.6 pH
At volume 23 mL

Moles of titrant (NaOH)

0.1 M =
23
Moles of titrant = 0.1 M 0.023 L
= 0.0023 moles
Since the moles of titrant is same with the moles of analyte, so we can calculate the
concentration of analyte.

Concentration of analyte (Acetic acid)

0.0023
= = 0.092 M
0.025

Since the pH is equals to pKa, pKa is equals to log Ka.

Calculation of Ka
pH = pKa = 8.6
pKa = -log Ka
Ka = 10- 8.6
= 2.5 10-9
Phosphoric acid
Volume of titrant added, mL pH

2.0 1.46
4.0 1.52
6.0 1.58
8.0 1.64
10.0 1.78
12.0 1.81
14.0 1.91
16.0 2.00
18.0 2.02
20.0 2.12
22.0 2.19
24.0 2.27
26.0 2.37
28.0 2.46
30.0 2.59
32.0 2.73
34.0 2.94
36.0 3.22
38.0 3.98
40.0 5.52
42.0 5.80
44.0 6.04
46.0 6.21
48.0 6.38
50.0 6.53
52.0 6.68
54.0 6.73
56.0 6.86
58.0 6.96
60.0 7.07
62.0 7.28
64.0 7.40
66.0 7.56
68.0 7.70
70.0 7.95
72.0 8.37
74.0 8.78
76.0 10.07
78.0 10.65
 Graph of pH versus volume titrant added
12

10

8
pH value

6
pH
4

0
0 10 20 30 40 50 60 70 80 90
volume of titrant added, mL

Equivalence point
(5.6 pH+ 4.0 pH) 2 = 4.8 pH
At the volume 39 mL.
As phosphoric acid has multiple equivalence points, we just choose the first as general
rule.
Moles of titrant (NaOH)

0.1 M =
39
Moles of titrant = 0.1 M 0.039 L
= 0.0039 moles
Since the moles of titrant is same with the moles of analyte, so we can calculate the
concentration of analyte.

Concentration of analyte (Phosphoric acid)

0.0039
= = 0.156 M
0.025

Since the pH is equals to pKa, pKa is equals to log Ka.

Calculation of Ka
pH = pKa = 4.8
pKa = -log Ka
Ka = 10- 4.8
= 1.5810-5
DISCUSSION
Based on these two experiments, the titration by using acetic acid (CH3COOH),
the volume of titrant (sodium hydroxide, NaOH) needed to neutralise the acetic acid is
lower than the volume of titrant to neutralise phosphoric acid (H3PO4). This is because,
from the pH meter, we could know that acetic acid is weaker than phosphoric acid. As
the initial reading for both acids, acetic acid is 3.18 pH value compared to phosphoric
acid is 1.46 pH value. For experiment titration of acetic acid, as the graph plotted
between pH values versus volume of titrant added, we could find the equivalence point
in order to find the moles of titrant. This is because, when we got the moles of titrant,
we could calculate the concentration of analyte. By using the formula:

From the formula, the concentration of analyte, acetic acid, we got 0.092 M.
However, in the lab manual, the concentration of acetic has given in 0.1 M. We think
that maybe there is something error happened to our experiment since we got the
colour change due to endpoint bright pink.
For concentration of analyte, phosphoric acid, we got 0.156 M. From these
results, we can conclude that acetic acid has a lower concentration than phosphoric
acid. To differentiate between acetic acid and phosphoric acid either they are
monoprotic or polyprotic, we can determine through the chemical writing. As we know,
acetic acid is CH3COOH and phosphoric acid is H3PO4. Apart from that, we know that
have only one hydrogen ion, H+ to combine with CH3COO-. Meanwhile, phosphoric
have three hydrogen ion, H+ to combine with PO43-. When acetic acid only contain one
hydrogen ion, is called as monoprotic acid. Then, phosphoric acid contain three
hydrogen ions which is called as polyprotic acid.
After calculate the concentration of analyte in order to find the value of Ka,
acetic acid got the value of Ka is 2.5 10-9. However, the Ka of acetic acid from the
table provided, should be 1.74 10-5. Also for phosphoric acid, the value of Ka in this
experiment is 1.5810-5. Meanwhile, from the table provided, Ka for phosphoric acid
should be 4.8 x 10-13 or 2.2 x 10-13. This is means that something wrong with my
calculation because we have did some mistake due carry out this experiment. When
the titration is reaching the end point, we did careless with titrated it over of titrant. So,
the colour change of the end point is bright pink. It should be light pink precisely.
Therefore, the data volume of titrant added ruined the calculation to get the exactly
value of Ka.