Practicum Report

General Biochemistry

Day/Date
Time
PJP
Asistant

: Thursday, 28th April 2016

: 15.00 17.30 WIB
: Syaefudin, SSi, MSi
: Listia Widyawati M
Rizqy Fachria

VITAMIN
Nishalini A/P Magis Paran

B04158008

FACULTY OF VETERINARY MEDICINE

BOGOR AGRICULTURAL UNIVERSITY
2016

INTRODUCTION

Vitamins are organic compounds which are very essential for complete health of
body but our body cannot synthesise it on its own. A very little amount of 13
vitamins must be obtained from diet or tablets for a healthy body. Deficiency of
vitamins can cause illness (Poedjiadi 1994).
Vitamins are classified by their solubility to fat soluble and water soluble
vitamins. Fat soluble vitamins are vitamins that dissolve in fat. It is because they
can be easily stored in fat tissues and liver in our body. This means they can be
accumulated and be saved for later use. The fat-soluble vitamins are A, D, E and
K. Water soluble vitamins are soluble in water and they cannot be stored in body
and will be expelled through urine because our body is a watery environment.
Thus, we have to replenish water soluble vitamins such as Vitamin B and Vitamin
C every day. There are eight B vitamins; they are vitamin B1, B2, B3, B5, B6, B7,
B9 and B12. Their names are thiamine, riboflavin, niacin, pantothenic acid,
pyridoxine, biotin, folate and cobalamin (Basset, J 1994).
The most important role of vitamin C is to increase immune system of body. It
also acts as an inhibitor of histamine, a compound that is released during allergic
reactions. Vitamin C is a powerful antioxidant which can neutralize harmful free
radicals and it helps in neutralizing pollutants and toxins. Vitamin C can
regenerate lipid soluble vitamin E in vitro (Packer et al. 1979; Sharma & Buettner
1993; Wijesunara & Berger 1994). Vitamin C is required for the biosynthesis of
collagen, L-carnitine. Collagen is an essential component of connective tissue,
which plays a vital role in wound healing. Collagen also functions as intercellular
cement and gives structures to muscles, vascular tissues, bones, tendons and
ligaments. Vitamin C also maintains health of teeth and gums from bleeding and
haemorrhaging. Moreover, it absorbs iron from diet and vital for overall eye
health. Vitamin C is a cofactor in at least eight enzymatic reactions, including
several collagen syntheses. Ascorbate is the anion of ascorbic acid which is
required for a range of essential metabolic reactions in all animals and plants. It is
synthesised internally by almost all organisms. Vitamin C is found in high
concentrations in immune cells, and is consumed quickly during infections. The
interaction of vitamin C towards immune system is not certain but it has been
hypothesized to modulate the activities of phagocytes, the production of cytokines
and lymphocytes, and the number of cell adhesion molecules in monocytes.
Structure of vitamin C is attached below as picture 1 (Harjadi, W 1990).
The objective of this experiment is to determine concentration of Vitamin C in
tablet and fruit.

Picture 1: Structure of Vitamin C

METHOD

Time and Place of Practicum

Vitamin practicum was held in Laboratorium B Biokimia, Departemen Biokimia,
Fakultas Matematika dan Ilmu Pengetahuan Alam, Institut Pertanian Bogor. The
time of practicum was at Thursday, 28th April from 1500 to 1730.
Materials and Apparatus
The apparatus used in the lab are beaker, Mohr pipette, bulb, a pipette,
Erlenmeyer flasks, burettes, spatula, water bath and a mortar.
The materials used are tablets of vitamin C, cold distilled water that has been
boiled previously, H2SO4 2N, 0.1N, iodine solution 0.1N thiosulfate solution,
starch solution, and UC1000.
Procedure
Determination of Vitamin C in the Tablet
50 mg of vitamin C tablet dissolved in 5 mL of cold distilled water that has
previously boiled. After that, 3 mL of H2SO4 2N and 10 ml of iodine solution 0.1
N added. Then, the solution titrated with 0.1 N thiosulfate solution and starch
solution used as an indicator. Then, blank titration was done. Blank titration was
done by adding 5 mL of distilled water to 3 mL H2SO4 2N and 10ml of 0.1 N
iodine solution and titrated with 0.1 N thiosulfate solution. Starch solution used as
an indicator.
Determination of Vitamin C in UC1000
5ml of UC1000 added to 3 mL H2SO4 2N and 10 ml of 0.1 N iodine solution.
Then the solution titrated with 0.1 N thiosulfate solution and starch solution used
as an indicator.

RESULTS AND DISCUSSION

Determining the concentration of vitamins in a material is done by indirect
Iodometry. 0.1N Sodium thiosulphate is used as the solution to be titrated (Harjadi
1986). H2SO4 solution was added so that Iodine solution does not undergo
oxidation when mixed with vitamin C solution which has oxidising properties.
Iodine reagent and starch are added as an indicator during the titration to
determine the concentration of vitamin C. Iodine will react to the presence of
excess titrant by changing it from red to pale yellow color. Starch with I2 form a
dark blue compound react to the presence of titrant by turning into pale yellow.
The endpoint was evident with a color change (Mulyono 2005).

Table 1 Concentration of Vitamin C in Tablet

Repetition
Control

Titrated Volume (ml)

Actual

Concentration of

Initial Final Used

Volume (ml)

Vitamin C (mg/tablet)

20.7

32.0

11.3

14.0

22.5

8.5

2.8

4.92

39.5

47.0

7.5

3.8

6.69

Average: 5.81mg
Calculation:
Concentration of vitamin C in a tablet = Actual Volume of Thiosulphate x 8.8mg
_________________________________
Volume of Vitamin C Solution
1st experiment = 2.8 x 8.8
5
= 4.92
2nd experiment = 3.8 x 8.8
5
= 6.69
The average concentration of vitamin C in tablet obtained from experiment is
5.81mg but the real concentration of vitamin C in the tablet is 50mg. The result is
far different from the actual. This could be because of the errors present during the
experiment. There could be mistakes happened during the titration maybe lesser
or more volume of Thiosulphate titrated.

Table 2 Concentration of Vitamin C in UC1000

Repetition
Control

Titrated Volume (ml)

Actual

Concentration of

Initial Final Used

Volume (ml)

Vitamin C (mg/tablet)

14.0

32.0

11.3

17.0

22.0

5.0

6.3

0.40

0.0

10.1

10.1

1.2

0.075

Average: 0.24mg

Calculation:
Concentration of vitamin C in UC1000 = Actual Volume of Thiosulphate x 8.8mg
_________________________________
Volume of Vitamin C Solution
1st experiment = 6.3 x 8.8
140
= 0.40
2nd experiment = 1.2 x 8.8
140
= 0.075
The average concentration of vitamin C in tablet obtained from experiment is
0.24mg but the real concentration of vitamin C in the tablet is 7mg. The result is
far different from the actual. This could be because of the errors present during the
experiment. There could be mistakes happened during the titration maybe lesser
or more volume of Thiosulphate titrated.
For the experiment using tablet, when H2SO4 added to the solution it turns to
brown. After titrated with Thiosulphate, the solution changes to blue. Then starch
added and titrated until yellow colour produced. For the experiment using the
UC1000, when H2SO4 added, the solution turned to brown. After titrated with
thiosulphate it turned to dark yellow. After that starch added and it was titrated
with thiosulphate until clear yellow coloured solution resulted.
Activity of vitamin C is generally destroyed by heat treatment of food, especially
when there are traces of metals such as copper, but it is resistant to freezing.
Vitamin C is probably one of the most unstable among the vitamins soluble in
water. In nature there are only L-form of ascorbic and dehydroascorbic
acid.Vitamin C is very soluble in water and methanol, and about five times lower
in ethanol. Ascorbic acid is well oxidized by air oxygen, especially in the presence
of heavy metal ions.In the absence of oxygen, ascorbic acid can withstand heat up
to 100C. Empirical formula of vitamin C is C6H8O6. Molecular weight of it is
176.1. Melting point is about 190C with decomposition.The appearance of
vitamin C is white to slightly yellowish crystalline powder, practically odorless,
with a strong acidic taste.
Scurvy is an avitaminosis resulting from deficiency of vitamin C because without
this vitamin, the synthesized collagen is too unstable to carry out its function.
Scurvy results to the formation of brown spots on the skin, spongy gums, and
bleeding from all mucous membranes. The spots are most found on the thighs and
legs, and a person with the ailment looks pale, feels depressed, and is partially
immobilized. In advanced scurvy there are open, suppurating wounds and loss of
teeth and, eventually, death.

CONCLUSION
Determining of vitamin C can be done by indirect titration. Concentration of
vitamin C in vitamin C tablets is 5.81 mg. The concentration of vitamin C in fruit
juice (UC 1000) is 0.24mg. Concentration of vitamin C in tablet is greater than the
concentration of vitamin C in UC1000.

REFRENCE
Basset, J. Dkk. 1994. Buku Ajar Vogel Kimia Analisis Kuantitatif Anorganik.
Jakarta.
Harjadi, W. 1990. Ilmu Kimia Analitik Dasar. Jakarta : Penerbit PT Gramedia.
Poedjiadi, Anna. 1994. DasarDasar Biokimia. Jakarta : Penerbit Universitas
Indonesia.