1.

INTRODUCTION
Carbohydrates play essential part in almost all the biological processes.
They are one of the four macromolecules vital to natural processes. They serve
as a major energy source for living organisms. For example, glucose which is the
main energy source in plants and animals. They also transport energy, serve as a
structural material and an originator for vast molecules.
A carbohydrate with the functional group aldehyde is classified as aldose
and ketose when it has ketone functional group. The subgroups under
carbohydrates are monosaccharides, oligosaccharides, polysaccharides and
glyconoconjugates. Glycogen is an example of a polysaccharide.
Glycogen, the major glucose storage polymer in animals, has a highly
branched structure which permits rapid release of glucose from glycogen stores,
e.g., in muscle cells during exercise.The ability to rapidly mobilize glucose is
more essential to animals than to plants. Glycogen is a very compact structure
that results from the coiling of the polymer chains.This compactness allows large
amounts of carbon energy to be stored in a small volume, with little effect on
cellular osmolarity. In this experiment, glycogen was isolated from the chicken
liver via precipitation.
1.1 Theories and Principles
Carbohydrates are carbon compounds that contain large quantities of
hydroxyl groups, and are also the most important sources of energy. They have
the basic general formula Cn(H2O)nand they are the most commonly found
organic compounds in living organisms. They are classified into several groups,
namely monosaccharides, disaccharides, and polysaccharides, depending on the
number of their monosaccharide units. In this experiment, chicken liver was the
material used to obtain glycogen, which is a polysaccharide. The glycogen from
chicken liver was extracted and purified through heating and precipitation. After
isolating the glycogen, it was further tested with different tests, such as Molischs
test and Iodine reaction.
For detecting the presence of carbohydrates in Molischs test, the solution
is first treated with a strong acid. This is for hydrolysing the carbohydrate to
monosaccharide.When water is removed from monosaccharides, a compound
called furfurol is formed. This furfurol is condensed to form a violet ring or other
colored compound. The compound furfurol is condensed with alpha nephthol or
other phenol present in molischs reagent.
On the other hand, Iodine reaction is a test for polysaccharides. Iodine
forms a coordinate complex between the helically coiled polysaccharide chain
and iodine centally located within the helix due to absorption. The color obtained
depends upon the length of the unbranched or linear chain available for complex
formation. For glycogen, it gives a reddish brown color.
1.2 Objective
The objective of this experiment is to isolate and perform the general tests to
know whether the glycogen is present in chicken liver.

2. METHODOLOGY.
The isolation of glycogen from the chicken liver is attained by using the
mechanism of precipitation. By mincing, grinding, and boiling the liver,
separation of the proteins from the glycogen found in the sample is elicited. After
isolating the glycogen, it was further tested with different tests, such as Molischs
test, in this test, Molisch reagent is mixed with a dilute solution of carbohydrate.
Concentrated sulfuric acid is introduced carefully, and a purple color develops at
the interface if a carbohydrate is present. The Iodine Test, on the other hand, is
used to identify glycogen and starch. Polysaccharides combine with iodine to
form a positive result as it produced reddish-brown coloration
2.1 Materials
The materials we used in this experiment are the following: Petri dish,
Test tube, Beaker, Glass rod, Mortar & Pestle, Filter paper, Water bath, Wash
bottle, Hot plate and Graduated cylinder. While the reagents we used are 0.1%
CHCOOH, 95% ethanol, 12N H2SO4 and 0.01 M I2.
2.2 Procedure
PART A: Extraction and Precipitation of Glycogen
A.1 For Isolation of Glycogen
Weigh 10 g of chicken liver using a triple beam balance and place on a
Petri dish. Cut the sample with the use of scissors. Transfer sample to a beaker
and pour 24 mL of distilled boiling water. Stir contents with a glass rod and boil
for four minutes using a hot plate to precipitate the proteins in the sample. Pour
the mixture into a mortar and use pestle to grind the sample thoroughly until no
lumps are visible. Add 6 mL distilled water and transfer the mixture into a
beaker. Heat the mixture in a boiling water bath for thirty minutes and, if
necessary, add distilled water to the mixture to avoid evaporation since glycogen
goes to the solution when heating. Filter solution using filter paper and separate
glycogen samples into three portions and transfer to test tubes.
*0.1% acetic acid may be added to improve precipitation of proteins during
heating of sample in boiling water bath.
A.2 For Glycogen Precipitation by Ethanol
Add five to ten drops of ethanol to 1 mL glycogen solution and observe
precipitation.
PART B: General Tests for Polysaccharides
B.1 For Molischs Test
Add a few drops of Molischs Reagent into 1 mL glycogen solution.
Carefully pour 2 mL conc. H 2SO4 down the side of a tube using a glass rod to
form a purple interface.
B.2 For I2 Reaction
Add a few drops of 0.01 M I2 into the sample solution measuring 1 mL.
A red colour produced indicates the presence of glycogen. Warm the mixture in
a water bath and observe if there is any change in colour. Cool and note the
result.

3. RESULTS & DISCUSSIONS

When glycogen was successfully extracted from the chicken liver, it was
isolated from impurities, specifically from proteins, by protein precipitation.
Precipitation of the
proteins, which was enhanced by 0.1% CHCOOH,
was
brought about by boiling the chicken liver with water. While heating, glycogen
was left soluble in the solution. The precipitate was separated from the solution
by the process of filtration. The successful extraction of glycogen was therefore
proved by acquiring positive results from the following test which tested the
presence of glycogen in the extracted solution glycogen precipitation by
ethanol, Molischs test and Iodine reaction.
Table 1. Results from testing the presence of glycogen
Test

Results

Glycogen
precipitation by
ethanol

(+) Flesh precipitate

Molischs Test

(+) Purple interface

I2 Reaction

(-) no reaction

Molischs test and iodine reaction test are assessments for the presence of
carbohydrates and polysaccharides. Glycogen is a carbohydrate and a
polysaccharide so it must be positive for these tests. Table 1 shows the results
from testing the presence of glycogen. The purple interface that forms in
Molischs test indicates that there is a presence of carbohydrates, it is due to the
formation of an unstable condensation product of beta-naphthol with furfural
(produced by the dehydration of the carbohydrate) so this give us a positive
result. For the iodine reaction test, our result shows no reaction. The mixture
should have been turned to reddish-brown for the presence of polysaccharide
because iodine forms coloured adsorption complexes with polysaccharides.
Therefore, the iodine reaction in this experiment was unsuccessful.
4. CONCLUSION
In this experiment, the general tests for polysaccharides were both
successful and unsuccessful. For the Molischs test it gives us a positive result
which indicates that there is a presence of carbohydrates. While in the iodine
reaction shows no any reaction. It may be due to the reagent we used. The
reagents for each test should have been assured to be uncontaminated
5. REFERENCEs
(1) http://www.ausetute.com.au/redsugar.html
(2) https://www.scribd.com/doc/124760418/Formal
(3) http://allmedicalstuff.com/molischs-test-qualitative-test-presencecarbohydrates/