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BIOCHEMISTRY MANUAL

Prepared by Admer Rey C. Dablio, v2012

Experiment 4
Score:

Isolation and General Tests of Glycogen


from Chicken Liver
Name: Joann H. Justiniane
Groupmates: Marla C. Basa
Ana Margarita L.
Baytion

Date Performed: July


2016
Instructors Signature:

14,

Objectives:

II

To verify the presence of glycogen from chicken liver.


To prove the existence of carbohydrate.
Chemicals:

III

0.1% CH3COOH
Molischs Reagent
0.01M Iodine

95% ethanol
12M H2SO4

Apparatus/Materials/Equipment:

Chicken liver
50 mL beaker
100 mL beaker
Stirring rod
3 Test tubes
Wire Gauze
Scissor
25 mL Graduated
cylinder

IV

Schematic Diagram of the Procedure:


Please write on a short bond paper.

Summary of Theory

Water bath
Test tube rack
Test tube holder
Hot plate
Bunsen Burner
Tong
Petridish
Mortar and Pestle

Z. Galewska, T. Gogiel, A. Malkowski, L. Romanowicz, K. Sobolewski, M.


Wolaska cited that Glycogen is a branched homopolysaccharide composed of D-glucose residues joined by glycosidic bonds. Straight sections contain glucose
residues joined by the -1,4 bonds, and branches are formed by -1,6- glycosidic
bonds. There is one branching for 8-10 glucose residues incorporated into the
linear chain. The high degree of glycogen chain branching helps fulfil its function.
It improves the solubility of the polysaccharide and increases the number of
terminal glucose residues, which may be released or may serve as acceptors for
the next molecules of this monosaccharide. Glycogen is the major glucose storage
polymer in animals, has a highly branched structure which permits rapid release
of glucose from glycogen stores. The ability to rapidly mobilize glucose is more
essential to animals than to plants. Glycogen is a very compact structure that
results from the curling of the polymer chains. The compactness of these polymer
chains allows large amounts of carbon energy to be stored in a small volume, with
little effect on cellular osmolarity. Glycogen acts as reservoir of glucose. The
amount of glucose stored as glycogen in dynamic equilibrium is rapidly changing

No part of this manual may be reproduced without written permission from the Chemistry Department
of the College of Arts and Sciences, Mindanao University of Science and Technology, Cagayan de Oro
City.

BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012

depending on the state of the body and the organs synthesizing it which is the
muscles and the liver.
The carbohydrate tests used in this experiment can be divided into two
classifications based on the mechanism of action which takes place on the
reagents used in the tests. The first test involves the use of dehydrating acids
followed by condensation of reagents. This is called the two-step analysis, which
often yield highly coloured results. The second classification
is making use of copper (II) ion-containing reagents. The copper (II) ions are
reduced to cuprous oxide copper (I) oxide by the carbohydrates present in the
samples.
The Molischs Test will show positive results for all carbohydrates, with
monosaccharides reacting much faster than disaccharides and polysaccharides.
In this experiment, glycogen was isolated from the chicken liver by
precipitation. Chicken liver is used in this experiment because it is a good source
of glycogen. Glycogen can be separated from other proteins by mincing, grinding,
and boiling the liver.
VI

Observations
Mass chicken liver = 6.0191 g

Sample Used
Sample Purchased from
Description of filtrate
Description of
precipitate

Chicken Liver
Shopwise Supermarket
Light yellow, stinky odor
Stinky, light brown

A. Molischs Test
Table A.1 Molischs Test
Sample
No. of drops of
Molischs
reagent
1 mL
5
Glycogen
B. Iodine Test
Table B.1 Iodine Test
Sample
No. of drops of
0.01M Iodine
soln
1 mL
10
Glycogen
VII

Observation

After adding 2 mL 12M


H2SO4

Peachy
solution

Hot and Purple ring


formation

Observation

After Heating

Slightly Yellow

No change

Analysis
According to Daniel Luzon Morris, Molischs Test was invented by
Hans Molisch, an Australian botanist. He said that this test was base from
the dehydration of carbohydrate with sulfuric acid to form furfural that will
react with alpha-napthol to give a violet or purple colored solution. He
further added that all carbohydrates monosaccharides, disaccharides, and
polysaccharides will give a positive reaction, and nucleic acids and
glycoproteins will also give a positive reaction, as all these compounds are
eventually hydrolyzed to monosaccharides by strong mineral acids.
Pentoses are dehydrated to furfural, while hexoxes are dehydrated to 5hydroxymethylfurfural. Either of these aldehydes, will condense with two
molecules of naphthol to form a purple-colored product, as illustrated
below by the example of glucose:

No part of this manual may be reproduced without written permission from the Chemistry Department
of the College of Arts and Sciences, Mindanao University of Science and Technology, Cagayan de Oro
City.

BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012

In Table A.1, Glycogen gave a positive result. This means that


glycogen is a homopolysaccharide carbohydrate. Glycogen was
dehydrated
by
H2SO4
to
form
hydroxymethylfurfural.
Hydroxymethylfurfural was then condensed with two naphthol to form a
purple colored product.
Morris also cited that Iodine Test was used to identify glycogen and
starch. Glycogen reacts with Iodine reagent to give a reddish-brown color
or blue-black color. It was thought that starch and glycogen form helical
coils. Iodine atoms can then fit into the helices to form a starch-iodine or
glycogen-iodine complex. Starch in the form of amylose and amylopectin
has less branches than glycogen. This means that the helices of starch are
longer than glycogen, therefore binding more iodine atoms. The result is
that the color produced by a starch-iodine complex is more intense than
that obtained with a glycogen-iodine complex.
In Table B.1, Glycogen didnt gave a positive result in Iodine
solution. The formation of reddish-brown or blue-black color indicates the
presence of glycogen. But in this experiment, the glycogen did not gave a
positive result. This kind of occurrence is because of method error. The
Iodine solution might have lost its function due to prolong storage in the
laboratory.
VIII

Conclusion
Therefore, glycogen will give a positive result in molischs reagent
and Iodine solution because it is a carbohydrate.
The experiment was not successful due to method error and human
error. The expected product was not formed because some of the reagent
might have lost its function to react with other samples.
For further experimentation of Isolation and General Tests of
Glycogen from Chicken Liver, the experimenter recommend to freshly
prepared all the chemicals that will be use in the experiment.

No part of this manual may be reproduced without written permission from the Chemistry Department
of the College of Arts and Sciences, Mindanao University of Science and Technology, Cagayan de Oro
City.

BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012

IX

References
1. Z. Galewska, T. Gogiel, A. Malkowski, L. Romanowicz, K. Sobolewski, M.
Wolaska, Biochemistry Workbook for students of the Faculty of
Medicine and the Faculty of Health Sciences Medical University of
Bialystok, Glycogen synthesis and degradation pp. 95-96, 2013.
2. D. L. Morris, Colorimetric determination of Glycogen, Disadvantages of
Iodine Method Received for publication July 15, 1998.

No part of this manual may be reproduced without written permission from the Chemistry Department
of the College of Arts and Sciences, Mindanao University of Science and Technology, Cagayan de Oro
City.

BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012

Appendix

Heating the Chicken


Liver

Addition of Molischs
reagent to a 1 mL solution

Filtration of
Glycogen

Addition of Iodine solution


to Glycogen

No part of this manual may be reproduced without written permission from the Chemistry Department
of the College of Arts and Sciences, Mindanao University of Science and Technology, Cagayan de Oro
City.