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Experiment 4
Score:
14,
Objectives:
II
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
Summary of Theory
Water bath
Test tube rack
Test tube holder
Hot plate
Bunsen Burner
Tong
Petridish
Mortar and Pestle
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
Peachy
solution
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
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
Addition of Molischs
reagent to a 1 mL solution
Filtration of
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.