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DEPARTMENT OF BIOLOGY

FACULTY SCIENCE AND MATHEMATICS

SULTAN IDRIS EDUCATION UNIVERSITY

SBL 1023

TECHNIQUE IN BIOLOGY AND BIOCHEMISTRY LABORATORY

Lab 2: Titration

AYU ILYANA BT ZULKIFLI


STUDENT`S NAME
( E20161014077)
EXPERIMENT`S NO LAB 2 : Titration

DATE/DAY/PLACE 21/11/2017 /TUESDAY/ B2-L3-MP 13

LECTURE`S NAME Professor Madya Dr. Shakinaz Binti Desa


1.0 Title : TITRATION

2.0 INTRODUCTION

Titration is a technique which a solution of the titrant was added to a flask which contain an
analyte, to know the unknown concentration of the analyte. In this experiment, we conduct two
experiment which include a weak acid, which is a titration of acetic acid and phosphoric acid,
which both the acid was titrate with sodium hydroxide.

The acid-base titration was monitored with an indicator or with a pH meter. In either case,
the goal is to determine the equivalence point of the titration. This is the point at which enough
titrant has been added to the analyte to just exactly neutralize the analyte.

In this experiment, knowledge of the equivalence point will be used to obtain information
about the acid dissociation constant, Ka, of the acid being titrated. When an indicator is used
in a titration, the colour change occurs at what is called the endpoint. If the indicator has been
properly selected, this point will be the same as the equivalence point. When a pH meter is
used, the pH of the solution is recorded as the titrant is added. The pH versus the volume of
titrant added can be plotted on what is called a titration curve. In this case the equivalence
point occurs at the point where very small additions of titrant cause a very rapid rise in the ph.

3.0 OBJECTIVE
 To determine the concentration of acetic acid
 To determine the concentration of phosphoric acid
 To obtain an equivalence point of titration that used to know the information about
the acid dissociation constant, Ka, of the acid being titrated.

4.0 EQUIPMENTS
 Phenolphthalein indicator
 Burette
 Pipette
 250ml beakers
 pH meter
 White paper towel
5.0 METHODOLOGY
A) Titration of monoprotic acid (acetic acid) with NaOH
1. 0.1M NaOH filled the burette. 25.00 mL of 0.1 M CH3COOH was pipetted and transferred
into a 250 mL beaker and 3-4 drops of phenolphthalein indicator was added. To obtain
best observed colour changes, the beaker was placed on a white paper towel
2. The solution was titrated by added the NaOH titrant in 2 mL increments. The beaker was
carefully swirled with each addition.
3. The coloured form of the phenolphthalein will began to stay for a while and then
disappeared. At this point the NaOH was added dropwise until the acetic acid is a 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. Then,
the pH probe was rinsed with distilled water and the probe tip placed to its vial.
5. During the titration, any colour changed observed was recorded. The pH and added NaOH
volume at that indicator’s endpoint used to estimate the target point.
6. The pH/volume data was transferred to an Excel file for analysis. The volumes stored in
the A column, while, the pH values in the B column of the spreadsheet, beginning in cells
A1 and B1.
7. The saved data in the Excel file used to prepare a plot of pH vs. Volume of NaOH added
and used to observe the equivalence point and half equivalence point.
8. The inflection point was determined to verify the equivalence point, by calculating the
change in pH per change in volume, ΔpH/ΔV, for each recorded volume. ΔpH/ΔV vs. the
volume of titrant added was plotted. Then, in cell C1 of the excel, (A1+A2)/2 was typed.
Then, cell C1 clicked and dragged down to the C cell that is on the same row as the last
filled cell in columns A and B. Column C now contained the average volume of titrant
between two readings.
9. In cell D1, (B2-B1)/ (A2-A1) was typed, which will calculated the ΔpH/ΔV values. Highlight
cell D1 and dragged to fill the remaining cells as done with column C. Column D now
contained the values of ΔpH/ΔV for each volume reading. By plotting ΔpH/ΔV versus the
volumes in column C, the inflection point was found.
10. Then, the volume of the half equivalence point can be found, once the volume at the
equivalence point is known. The pH at this volume can be determined at the titration
curved. Thus, it shown the pKa and hence Ka for acetic acid been calculated.
B) Titration of polyprotic acid (phosphoric acid) with NaOH
Repeat all the steps 1 to 10 with phosphoric acid.

6.0 DATA AND RESULT

DATA PART A

Volume of NaOH (ml) pH


0.00 2.64
2.00 4.35
4.50 4.77
6.10 4.89
8.00 5.05
10.00 5.20
12.00 5.30
14.00 5.54
16.00 5.67
18.00 5.77
20.00 6.04
22.00 6.41
24.00 7.56
24.20 10.83
26.00 12.07
28.00 12.27
30.00 12.40
Table 1.0: Titration of monoprotic acid (acetic acid) with NaOH
DATA PART B

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 2.0: Titration of polyprotic acid (phosphoric acid) with NaOH.

Data credit to Jasmi Arif


GRAPH

GRAPH: PART A

pH vs volume of NaOH
14

12.4
12.27
12.07
12

10.83

10

pH at eq.(9.195 pH)

8
7.56
pH

6.41
6.04
6 5.67 5.77
5.54
5.2 5.3
5.05
pH = pKa = 4.77 4.77 4.89
4.35

2.64

0
0 5 10 15 20 25 30 35
Volume of NaOH
VNaOH = 24.1 mL

Graph 1.0: Titration of monoprotic acid (acetic acid) with NaOH


GRAPH: PART B

pH vs volume of NaOH
14

12 11.55
11.44
11.27
11.04
pH at eq = 9.5 10.72
= - log [H+]
[H+] = 3.16 X 10 -10 9.93
10
9.2

8.66
8.22
pH = pKa = 7.2 8.01
7.84
8 ka = 6.31 x 10 -8 7.7
7.59
7.49
7.39
7.29
7.18
7 7.1
6.87
6.84
6.73
pH

6.63
6.48
6.25
pH = 4.85 5.87
6
pH = - log [H+]
[H+] = 1.41 x 10 -5
4.61

4 3.69
pH = pKa = 2.15 3.35
ka = 7.08 x 10 -3 3.16
3.01
2.89
2.78
2.68
2.58
2.5
2.42
2.34
2.2
2.15 2.27
2.24
1.9722.03
2

Volume at eq.
= 38.5 ml
0.13
0
0 10 20 30 40 50 60 70 80 90 100
Volume of NaOH (mL)

Graph 2.0: Titration of polyprotic acid (phosphoric acid) with NaOH


RESULT

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

Diagram 1.0: Titration NaOH solution with acetic acid solution

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

Diagram 2.0: Titration NaOH solution with phosphoric acid solution

7.0 DISCUSSION
In this experiment, we have done two investigation on two analyte which is acetic acid
and phosphoric acid with the titrant, which is the sodium hydroxide (NaOH). The titration was
made until reach the equilibrium point and end point where, when the solution in the flask, the
analyte turn from colourless to light pink in colour .

By the experiment that had been conduct, the volume of analyte that was used was
same, 25 mL, but the volume of titrant which is used to titrate the analyte was differ, which is
for equivalent point for acetic acid is 24.1 mL, while for the phosphoric acid is 38.5 ml. This is
because acid dissociation in the water was differ. The phosphoric acid was higher than acetic
acid. The phosphoric acid can donated two or three protons ions respectively.

Besides, based on the both graph that had been plotted, both graph was different with
each other, by the s-shaped curve. This is the titration curve. The acetic acid have only one s-
shaped curve. While, the phosphoric acid solution titrate with Naoh contained 2 S-shaped.
This is because both has difference in donation of hydrogen ions. According the graph in 1.0
and 2.0, we can determined what type of acid is. The acetic acid is a monoprotic acid, which
acid that can donate only one hydrogen ions. While, phosphoric acid is a polyprotic acid, acid
that can donate multiple hydrogen ions. Monoprotic and polyprotic acid can be identified from
the graph of titration according to the number titration curves.

The acetic acid dissociation:

CH3COOH ⇆ CH3COO- + H3O+

The phosphoric acid dissociation

H3PO4 ⇆ H2PO4- + H+

Furthermore, the titration was started with calibration of the pH meter. This process is
important because we need to standardize the buffer at pH in range 4, 7 and 10. With the
calibration of the pH meter, it can maintained accuracy. To make a calibration curve at least
three standards are needed. Without the standardized pH buffer to calibrate the meter the
results will be inaccurate and useless. Because, buffer solutions are used to calibrate pH
meters because they resist changes in ph. The function of the pH meter is to measure H+
activity (acidity or alkalinity) in the solution.
8.0 CALCULATION

Calculation part A: Titration NaOH solution with acetic acid solution

1) Mole of titrant (NaOH)

= 0.0241 x 0.1 (M)

=2.41 X 10-3 mol NaOH

Mole of titrant = mole of analyte

Concentration of analyte

= 2.41 x 10 -3 mol NaOH = 0.0964 M oxalic acid

0.025 L

2) pKa =pH= 4.77

3) pKa = -log Ka

Ka = 10-4.77

Ka = 1.698 x 10-5

Conclusion part B: Titration of polyprotic acid (phosphoric acid) with NaOH

Vequivalence point 1 = amount of titrant added to reach equivalence point

= 38.5 ml

Therefore,

𝒎𝒐𝒍𝒆𝒔 𝒐𝒇 𝒕𝒊𝒕𝒓𝒂𝒏𝒕 = 𝒗𝒐𝒍𝒖𝒎𝒆𝒔 𝒐𝒇 𝒕𝒊𝒕𝒓𝒂𝒏𝒕 (𝑳) × 𝒄𝒐𝒏𝒄𝒆𝒏𝒕𝒓𝒂𝒕𝒊𝒐𝒏 𝒐𝒇 𝒕𝒊𝒕𝒓𝒂𝒏𝒕 (𝑴)

1𝐿 𝑚𝑜𝑙
= 38.5 𝑚𝑙 𝑥 𝑚𝑙 𝑥 0.1
1000 𝑙

= 0.00385 𝑚𝑜𝑙

Moles of analyte = moles of titrant = 0.00385 mol

(𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑝ℎ𝑜𝑠𝑝ℎ𝑜𝑟𝑖𝑐 𝑎𝑐𝑖𝑑)


𝑐𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝑎𝑐𝑒𝑡𝑖𝑐 𝑎𝑐𝑖𝑑 = 𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑝ℎ𝑜𝑠𝑝ℎ𝑜𝑟𝑖𝑐 𝑎𝑐𝑖𝑑
𝑚𝑜𝑙 1000𝑚𝑙
= 0.00385 25
𝑚𝑙 𝑥 1𝐿

= 0.154 M

𝑽𝒆𝒒𝒖𝒊𝒗𝒂𝒍𝒆𝒏𝒄𝒆 𝒑𝒐𝒊𝒏𝒕 𝟐

= 𝒂𝒎𝒐𝒖𝒏𝒕 𝒐𝒇 𝒕𝒊𝒕𝒓𝒂𝒏𝒕 𝒂𝒅𝒅𝒆𝒅 𝒕𝒐 𝒓𝒆𝒂𝒄𝒉 𝒆𝒖𝒊𝒗𝒂𝒍𝒆𝒏𝒕 𝒑𝒐𝒊𝒏𝒕 = 𝟕𝟖. 𝟓 𝒎𝒍

Therefore,

𝒎𝒐𝒍𝒆𝒔 𝒐𝒇 𝒕𝒊𝒕𝒓𝒂𝒏𝒕 = 𝒗𝒐𝒍𝒖𝒎𝒆 𝒐𝒇 𝒕𝒊𝒊𝒕𝒂𝒏𝒕 (𝑳)𝒙 𝒄𝒐𝒏𝒄𝒆𝒏𝒕𝒓𝒂𝒕𝒊𝒐𝒏 𝒐𝒇 𝒕𝒊𝒕𝒓𝒂𝒏𝒕 (𝑴)

1𝐿 1𝑚𝑜𝑙
78.5 𝑚𝑙 𝑥 𝑥 0.
1000𝑚𝑙 𝐿

= 0.00785 𝑚𝑜𝑙

Moles of analyte= moles of titrant = 0.00785 mol

𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑝ℎ𝑜𝑠𝑝ℎ𝑜𝑟𝑖𝑐 𝑎𝑐𝑖𝑑


𝑐𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝑎𝑐𝑒𝑡𝑖𝑐 𝑎𝑐𝑖𝑑 =
(𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑝ℎ𝑜𝑠𝑝ℎ𝑜𝑟𝑖𝑐 𝑎𝑐𝑖𝑑

0.00785 𝑚𝑜𝑙 1000𝑚𝑙


= 𝑥
25 𝑚𝑙 1𝐿

= 0.314 𝑀

𝒑𝑯 𝒂𝒕 𝒆𝒒𝒖𝒊𝒗𝒂𝒍𝒆𝒏𝒄𝒆 𝒑𝒐𝒊𝒏𝒕 𝟏 = 𝟒. 𝟖𝟓

𝑝𝐻 = −𝑙𝑜𝑔 [𝐻+]

[𝐻+] = 1.41 𝑥 10 − 5

𝑝𝐻 = 𝑝𝐾𝑎

𝑝𝐾𝑎 = −𝑙𝑜𝑔 𝑘𝑎

𝑝𝑘𝑎 = 2.15

𝑘𝑎 = 7.08 𝑥 10 − 3

𝒑𝑯 𝒂𝒕 𝒆𝒒𝒖𝒊𝒗𝒂𝒍𝒆𝒏𝒄𝒆 𝒑𝒐𝒊𝒏𝒕 𝟐 = 𝟗. 𝟓

𝑝𝐻 = −𝑙𝑜𝑔 [𝐻+]

[𝐻+] = 3.16 𝑋 10 − 10
𝑝𝐻 = 𝑝𝑘𝑎

𝑝𝑘𝑎 = −𝑙𝑜𝑔 𝑘𝑎

𝑝𝑘𝑎 = 7.2

𝑘𝑎 = 6.31 𝑥 10 − 8

9.0 REFLECTION

In 21
November 2017, we had done the titration lab. Through this lab, we conclude that if the higher
the pH value, the higher the amounts of volume NaOH were used until it reached the
equivalent point. Then, after it approach the end point the volume is high again as, the pH is
higher. Based on the result, I can conclude that the volume of titrant required to completely
neutralize an acid depends on the amount of hydrogen ions contained in an acid. Besides, the
difference between the acetic acid and phosphoric acid is in the donation of the hydrogen ions.
This is the reason why the acetic acid required less volume of NaOH than phosphoric acid, to
reach the equilibrium point. The concentration of the acids was determined by doing some
calculation. Lastly, in a titration, the parallax error must been avoided for more precise and
accurate results. For example, we must take accurate reading of the data. Besides, we must
use a right technique such as drop the titrant slowly and be patient until it reach the equivalent
points. Furthermore, we must sure all the equipment were clean, not broken and in a good
condition. Thus, that are a few precaution that we can take through this titration laboratory.

10.0 REFERENCE
1) 1. Lab manual
2) 2. Lecture notes