LABORATORY REPORT

TECHNIQUE IN BIOLOGY AND BIOCHEMISTRY LABORATORY

(SBL1023)

STUDENT NAME ALISYA SYAZRIN ADILA BINTI

MOHD NASIR
REGISTRATION NO. E20161015898
GROUP B
EXPERIMENT EXPERIMENT 2: TITRATION
LECTURER NAME ASSOCIATE PROF. DR SHAKINAZ
DESA
 PRACTICAL 2: TITRATION

INTRODUCTION
An acid/base titration can be 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 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 color change occurs at what is called end point. 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 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.

OBJECTIVES
1. To determine the equivalence point of the titration.
2. To obtain information about the acid dissociation constant, Ka, of the acid being titrated.

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

PROCEDURE
1. 0.1 M NaOH was filled in the buret. Pipette 25 ml of 0.1 M CH3COOH into a 250 ml
beaker and 3-4 drops of phenolphthalein indicator was added. The beaker was placed on a
white paper towel to best observe color change.
2. The solution was titrated by adding the NaOH titrant in 1-2 ml increments. The beaker was
swirled carefully with each addition.
3. The colored from the phenolphthalein will begin to stay for a while and then disappeared.
At this point the NaOH was added dropwise until the acetic acid is a very light color. This is
the end point for phenolphthalein.
4. The pH of the solution in the beaker was measured and recorded at this end point. Then
the pH probe was rinsed with distilled water and replace the probe tip into its vial.
5. The color change was recorded during the titration. The pH and NaOH volume was added
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. The volumes in the
A column was stored 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
observed the equivalence point and half equivalence point.
8. To verify the equivalence point, the inflection point was determined by calculated the
change in pH per change in volume, ∆pH/∆V, for each recorded volume. Plot ∆pH/∆V vs the
volume of titrant was added. To perform this, the volume was checked 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 ∆pH/∆V values, highlight cell D1
and drag to filled the remaining cells with column C. Column D now contains the values of
∆pH/∆V for each volume reading. The inflection point is found by plotting ∆pH/∆V vs the
volumes in column C.
10. Once the volume at the equivalence points was known, the volume of the half
equivalence point was founded. Using the titration curve, the pH at this volume can be
determined. Thus pKa and hence Ka for acetic acid can be calculated.

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

Volume of NaOH (ml) pH

0.00 2.93
2.00 4.00
4.00 4.02
6.00 4.25
8.00 4.46
10.00 4.57
12.00 4.72
14.00 4.83
16.00 5.00
18.00 5.24
20.00 5.60
22.00 6.36
24.00 11.13
26.00 11.57
28.00 12.32
30.00 12.98
32.00 13.68
 Titration of acetic acid
pH at equivalence
16
point
+ 14
pH= -log [H ]

+
12
9.20= -log [H ]
PH

10
+
-9.20= log [H ]
8 EP= 9.20
+ (-9.20)
[H ]= 10
6
+ -10
[H ]= 6.30 x 10 4
2
0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
VOLUME OF TITRANT (ML)

pH = pKa
Volume
equivalence
point

Graph 1: Titration curve of acetic acid

CALCULATIONS

Vep = amount of titrant added to each EP

Therefore,
Moles of titrant = Vep (L) x concentration of titrant (M)

Vep = 23.00 mL
1𝐿
= 23.00 mL x 1000 𝑚𝐿

= 0.023 L

Moles of titrant = 0.023 L x 0.1 M

= 0.0023 moles of titrant used to reach the equivalence point
 Moles of titrant = moles of analytes

So, moles of analyte = 0.0023 mol

𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑎𝑛𝑎𝑙𝑦𝑡𝑒 (𝑚𝑜𝑙)

Concentration of analyte =
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑎𝑛𝑎𝑙𝑦𝑡𝑒 (𝐿)

0.0023 𝑚𝑜𝑙
Concentration of analyte = 1𝐿
0.025 𝐿 25.00 mL x
1000 𝑚𝐿
= 0.092 mol/L
= 0.025 L

pKa = -log Ka

pH = pKa

Then,
pKa = 4.80
pKa = -log Ka

4.80 = -log Ka
Ka = 10(-4.80)
Ka = 1.58 x 10-5
B. Titration of polyprotic acid (phosphoric acid) with NaOH

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
 Titration of phosphoric acid
14

pH at equivalence 12
point
10
+
pH= -log [H ]
8
+
PH

4.61= -log [H ]

+
6
-4.61= log [H ]
4 EP= 4.61
+ (-4.61)
[H ]= 10
2
+ -5
[H ]= 2.45 x 10
0
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45
VOLUME OF TITRANT (ML)

Volume
pH = pKa
equivalence
point

Graph 2: Titration curve of phosphoric acid

CALCULATIONS

Vep = amount of titrant added to each EP

Therefore,

Moles of titrant = Vep (L) x concentration of titrant (M)

Vep = 38.00 mL
1𝐿
= 38.00 mL x 1000 𝑚𝐿

= 0.038 L

Moles of titrant = 0.038 L x 0.1 M

= 0.0038 moles of titrant used to reach the equivalence point
 Moles of titrant = moles of analytes

So, moles of analyte = 0.0038 mol

𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑎𝑛𝑎𝑙𝑦𝑡𝑒 (𝑚𝑜𝑙)

Concentration of analyte =
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑎𝑛𝑎𝑙𝑦𝑡𝑒 (𝐿)

0.0038 𝑚𝑜𝑙
Concentration of analyte = 1𝐿
0.025 𝐿 25.00 mL x 1000 𝑚𝐿
= 0.152 mol/L
= 0.025 L

pKa = -log Ka

pH = pKa

Then,
pKa = 2.27
pKa = -log Ka

2.27 = -log Ka
Ka = 10(-2.27)
Ka = 5.37 x 10-3
DISCUSSION
Titration is a technique for determining either the concentration of a solution of unknown
molarity or the number of moles of a substance in a given simple. A chemical reaction is
used for this purpose, and the reaction must be fast, be complete, and have a determinable
end point. The reactions of strong acids and bases generally meet these criteria, and acid-
base titrations are among the most important examples of this technique. In this experiment,
the sample is acetic acid and phosphoric acid. The titration curve for the titration of acetic
acid is shown in Graph 1. Acetic acid which is a weak acid has been titrated with a 0.1 M
solution of NaOH solution. At the first point on the curve, the pH value is 2.93. The pH at this
point is the value of pure acetic acid. During the titration, the concentration of acid will
decrease. 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. Hence, the pKa for acetic acid also be 4.80 (by applying the Henderson –
Hesselbalch equation, pH – pKa). So, the ionization constant, Ka, is 1.58 x 10-5.
There are some mistake like use more than acid volume to titrate solution of NaOH. The
volume has pass the end point, so the volume uses is much more than needed. So, the
colour has changed to dark purple from light pink. To overcome this problem, we can titrate
slowly and shake the volumetric flask for 30 seconds when the solutions show changing in
colour from colourless to light pink. The other reasons is the using of volumetric flask that’s
had affect. To overcome this problem, we must make sure the volumetric flask is totally
clean and dry. That’s problem had affect our result. That’s why our result is not precise and
accurate to the correct value.
There is some recommendation for this experiment. First, we must ensure that there are no
bubbles trapped at the tip of the burette during the filling of NaOH solution. Second, we
,must avoid from make a parallex error for most of the measured content. Third, the NaOH
solution is titration drop by drop when there are change colour of the acid in the conical flask.
Lastly, we must take each measurement must be sharply as can so that the final result being
better.

CONCLUSION
An indicator in the erlenmeyer flask to indicate end-point. The end-point is where indicator
starting colour changes to another colour. For example the phenolphthalein changes from
colourless (acid) to light pink (base). The equivalence point is very close to this change in
colour where, the concentration of OH- ions is equal to the concentration of hydrogen ions:
[OH-] = [H+]. The [H+] in the erlenmeyer flask is determined by the volume NaOH dispensed
from the burette when reached the equivalence point which is approximately equal or close
to the end-point. The pH of having a strong base and a concentrated acid reacting together
to go to a complete reaction is approximately 7. If the titrant volume and molarity is known,
and the volume of the acid put into the flask is known, we can calculate the molarity of the
phosphoric and acetic acid. M1V1 = M2V2, rearrange for M2. Molarity of sulfuric acid =
(Molarity of NaOH x dispensed volume of NaOH / volume of acid in flask). NaOH is a strong
base and usually ionizes 99% or more in solution. H2SO4 is also a strong acid but is
diprotic. Has 2 hydrogens or 2 protons that can be dissociated in a chemical reaction if
enough base is present.
REFERENCES
1. Quantitave Analysis by Using Redox Titration, Alberta Education, 1996, Evanston AGC
United Learning.
2. http://www.easychem.com.au/the-acidic-environment/acid-base-definitions/titration-
technique
3. Fundamental of Chemistry, David E. Goldberg, McGraw Hill, 303.