Introduction

Glycogen is a branched polysaccharide composed of α- D-glucose residues joined

by glycosidic bonds.

Glucose residues joined by the α-1,4 bonds, and branches are formed by α- 1,6-
glycosidic bonds..

High degree of glycogen chain branching helps to improves the solubility

Glycogen is the major glucose storage polymer in animals.

The compactness of these polymer chains allows large amounts of carbon energy
to be stored in a small volume.
Principle
The concentration of glycogen is higher in the liver (10% by weight) than in
muscle (2% by weight).

The carbohydrate tests can be divided into two classifications based on the
mechanism of action .

The first test involves the use of dehydrating acids followed by

condensation of reagents.

The Molisch’s Test should show positive results for all carbohydrates.

Glycogen can be separated from other proteins by mincing, grinding, and

boiling the liver
Materials
 0.1% Methanol  glass Stirring rod  Test tube holder

 95% ethanol  3 Test tubes  Hot plate

12M H2SO4  Wire Gauze  Bunsen Burner

 Molisch’s Reagent  Scissor  Petri plates

 Chicken liver  25 mL Graduated cylinder  Mortar and Pestl

 50 mL beaker  Water bath  0.01M Iodine

 100 mL beaker  Test tube rack

Methods
• Wash chicken liver with distilled water in a Petri plate

• Take 20 grams of washed chicken liver in mortar and pestle

• Add 50 mL of phosphate buffer (7.4 pH)

• Mince the sample into a homogenize sample to even and smooth

consistency and then transfer to falcon tubes 25 mL each.

• centrifuge at 3000 rpm for 10 minutes

• Collect supernatant in a test tube

• Add 1 mL of 10% acetic acid

Methods
• Boil the sample in water bath for five minutes

• Let it cool down and then centrifuge at 3000 rpm for five minutes

• Collect supernatant to a new test tube and place it in a refrigerator for 30 min

• Add 5 mL of Absolute ethanol and place test tube in refrigerator for another 60
min

• Centrifuge at 3000 rpm for 10 minutes

• Discard supernatant and Collect precipitate (Glycogen)

• Dissolve the precipitate in 5 mL distilled water.

Methods
Hydrolysis

• Take 3 mL of Glycogen solution and add equal amount of 6M HCl.

• Boil the test tube in a boiling water bath for 30 minutes.

• Add 3 mL of concentrated ammonium hydroxide

Molisch test

• Take 10 drops of hydrolyzed sample and add 10 drops of Molisch reagent

• Mix thoroughly

• TILT the test tube carefully and add one mL of H2SO4 slide down the side of the
test tube to form a layer at the bottom and observe the color
Results & interpretation
Description Result

Chicken liver Fresh, radish

Homogenized mixture Light yellow, stinky odor

Precipitation Stinky, white to light brown

After refrigeration White precipitates

Molisch test Hot and Purple ring formation

Iodine test reddish-brown or blue-black

color
Conclusion
Glycogen will give a positive result in molisch’s reagent and Iodine solution
because it is a carbohydrate.