DEPARTMENT OF BIOLOGY

FACULTY OF SCIENCE AND MATHEMATICS

SULTAN IDRIS EDUCATION UNIVERSITY

SBL 1023

TECHNIQUE IN BIOLOGY AND BIOCHEMISTRY LABORATORY

EXPERIMENT 2 : TITRATION

NAME: NOR HAIZATUL BINTI SABIDI

NO MATRIC: E20161015640

GROUP : B

LECTURE NAME : PROFESOR MADYA DR. SHAKINAZ BINTI DESA

Title

Experiment 2: Titration

Objective

1. To determine the equivalence point of the titration

2. To determine the concentration of analyte.

Materials

1. Phenolphthalein indicator
2. Buret
3. Pipette
4. 250mL beakers
5. pH meter
6. White papert towel

Methods

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

1. The buret is filled with 0.1M NaOH. Pipet 25.00 mL of 0.1M CH3CHOOH into a
250mL of beaker and 3-4 of phenolphthalein indicator is dropped inside the beaker.
Beaker is placed on a white paper towel and observed the color changes.
2. The solution is titrate by adding the NaOH in 1-2 mL increments. The beaker is swirl
with each addition to mix the solution.
3. The color form of the phenolphthalein is begin to stay for a while and then disappear.
At this point, the NaOH is dropwise until the acetic acid is very light color. This show
the end point of phenolphthalein.
4. The pH of the solution is measured and recorded in the beaker at this point. Then the
pH probe is rinsed with distilled water and the probe tip is placed into its vial.
5. Any color change observed is recorded during this titration. The pH and added NaOH
volume at that indicator’s endpoint should be used to estimate the target point when
conducting the following procedure.
6. The pH/volume data is transferred to an excel file for the analysis. The volumes is
stored in the A column and the pH values in the B column of the spreadcheet,
beginning in cells A1 and B1.
7. The graph of pH vs Volume of NaOH is prepared using the data saved in Excel file to
observed the equivalence point and half equivalence point.
8. To verify the equivalence point, the inflection point is determined by calculating the
change in pH per change in volume, ∆pH/∆V, for each recorded volume.
9. 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 Ka for acetic acid can be calculated.
 B. Titration of polyprotic acid (phosphoric acid) with NaOH

Repeat all the steps above with phosphoric acid

Results

Acertic acid with Naoh

Table 1: Volume of Titrant and pH

Volume of titrant pH
0 2.93
2 4.00
4 4.02
6 4.25
8 4.46
10 4.57
12 4.72
14 4.83
16 5.00
18 5.24
20 5.60
22 6.36
24 11.13
26 11.57
28 12.32
30 12.98
32 13.68

Acetic Acoid
16
14
12
10
pH

8 ph
6
2 per. Mov. Avg. (ph)
4
2 Linear (ph)
0
0 5 10 15 20 25 30 35
volume of NaOH (mL)

Graph 1
Phosphoric acid with NaOH
Table 2: Volume of Titrant and pH
Volume of Titrant pH
0 1.30
2 1.97
4 2.00
6 2.03
8 2.15
10 2.20
12 2.24
14 2.27
16 2.34
18 2.42
20 2.50
22 2.58
24 2.68
26 2.78
28 2.89
30 3.01
32 3.16
34 3.35
36 3.69
38 4.61
40 5.87
42 6.25
44 6.48
46 6.63
48 6.73
50 6.84
52 6.87
54 7.00
56 7.10
58 7.18
60 7.29
62 7.39
64 7.49
66 7.59
68 7.70
70 7.84
72 8.01
74 8.22
76 8.66
78 9.20
80 9.93
82 10.72
84 11.04
86 11.27
88 11.44
90 11.55
 Phosphoric Acid
14

12

10

8
pH

pH
6
2 per. Mov. Avg. (pH)

4 Linear (pH)

0
0 20 40 60 80 100
volume of NaOH (mL)
Graph 2

Calculation

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

Moles of titrant = moles of analytes

𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑎𝑛𝑎𝑙𝑦𝑡𝑒𝑠,𝑚𝑜𝑙
Concentration of analytes =
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑎𝑛𝑎𝑙𝑦𝑡𝑒𝑠,𝐿

pKa = -log Ka
Acetic acid

Volume of NaOH (L) 0.0232 L

pH at eq. point 9.1
pKa 4

Moles of titrant = 0.0232 L x 0.1 M

= 2.32x10-3 mol

Moles of analytes = 2.32x10-3 mol

2.32x10−3 mol
Concentration of analytes =
0.025 𝐿

= 0.0928 mol/L

pKa = 4

pKa = -log Ka

4 = -log Ka

Ka = 10(-4)

Ka = 1x10-4

Phosphoric acid

Volume of NaOH (L) 0.0382 L

pH1 at eq. point 5
pKa1 2

Moles of titrant = 0.0382 L x 0.1 M

= 3.82x10-3 mol

Moles of analytes = 3.82x10-3 mol

3.82x10−3 mol
Concentration of analytes =
0.025 𝐿

= 0.1528 mol/L
pKa1 = 2

pKa1 = -log Ka1

2 = -log Ka1

Ka1 = 10(-2)

ka1= 0.01

Volume of NaOH (L) 0.0785 L

pH2 at eq. point 9.6
pka2 6.8

Moles of titrant = 0.0785 L x 0.1 M

= 7.85x10-3 mol

Moles of analytes = 7.85x10-3 mol

7.85x10−3 mol
Concentration of analytes =
0.025 𝐿

= 0.314 mol/L

pKa2 = 6.8

pKa2 = -log Ka2

6.8 = -log Ka

Ka = 10(-6.8)

Ka2= 1.585x10-7

Discussion

Generally, experiment of titration is to determine the concentration of an unknown

solution. A solution of known volume of titrant is used to determine the concentration of an
unknown analyte. Often, phenolphthalein indicator is used to signal the end of the reaction in
this experiment of titration. Titrant and analyte is a pair of acid and base.

Phenolphthalein indicator is a molecular substance that changes color when it comes

into contact with acids and bases and will be used to show when the reaction has completely
neutralized or the end point is reach. Phenolphthalein is colorless when in an acidic solution
and turns pink in a basic solution. Once enough base is added to neutralize the acid, the
reaction reaches the equivalence point. The point at which the moles of base is equal to the
moles of acid. The equivalence point can be used to determine the initial concentration of an
acid. The goal of the titration is to get as close as possible to the equivalence point by careful
addition of the base.

Based on the titration curve of acetic acid in graph 1, the pH value at equivalence
point is 9.1. Since the acetic acid is a monoprotic acid, it has only one pKa point which is 4.
So, the ionization constant, Ka, is 1x10-4.

Based on the titration curve of phosphoric acid in graph 2, the pH value at

equivalence point is 5 and 9.6. phosphoric acid is a polyprotic acid, so it has 2 pKa point
which is 2 and 6.8. the ionization constant ,Ka1 is 0.01 and for Ka2 is 1.585x10-7. As an acid,
a polyprotic acids have a very small acid dissociation constant (Ka), which measures the
strength of the 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. Next, we are not precisely added 2 mL of 0.2 M NaOH for every step of titration
process. From this experiment, there have some tips. First, 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.

Reference

 Chemisty Libretexts. Titration. 12 July 2016. From

https://chem.libretexts.org/Demonstrations_and_Experiments/Basic_Lab_Techniques/
Titration

 SCRIBD. Labreport titration. 20 January 2018 from

https://www.scribd.com/doc/21298837/Experiment-3

Reflection

Titration experiment is experiment that test the patience of student while handling it.
It take a time for the solution to turn pink. And they have to make sure that the pink color is
paler to get an accurate result. The best part is we happy when we get the paler solution. The
worst part is we have to be patience and get a strong hand in doing this experiment. In the
future, i wish i can try doing this experiment with other type of indicator because i want
learn more about the indicator that use to detect the neutralization oh acid-base.