Name: Missy Arabella C.

Pame Date Performed: June 20, 2021

Course and Year: Bachelor of Science in Nrsing1 Name of Lab Instructor: Ma’am Guadalupe Oracion

GLYCOGEN
Experiment No. 4

I. OBJECTIVES:
1. To prepare and study the properties of glycogen
II. APPARATUS: Bunsen burner, test tubes, mortar and pestle, evaporating dish, spatula, beaker
(250 ml), stirring rod, erlenmeyer flask (125 ml), funnel, test tube holder, test tube brush, dropper
III. MATERIALS: Oyster’s meat, sand, acetic acid, sodium chloride, starch solution, iodine solution,
Fehling’s solution A & B, concentrated hydrochloric acid, 6 N sodium hydroxide.
IV. PROCEDURE:
Preparation of Glycogen:
Mince one or two pieces of oyster meat and grind together with sand in a mortar. Transfer the
mixture to an evaporating dish, add about 40 ml of water and boil for 20 minutes. Replenish the water
lost by evaporation. Note the opalescence of the solution. Filter. Wash the residue on the filter paper
twice with two 10 ml. Portions of water adding the washings to the first filtrate. Heat the combined
filtrates to boiling and filter again. Concentrate the filtrate to a small volume by boiling; faintly acidify
with acetic acid and filter off any coagulated protein. Use the opalescent filtrate (glycogen solution)
for the following tests:
1. To 5 ml of the glycogen solution add 3-5 drops of sodium chloride and several drops of iodine
solution. Note the color and compare it with the color produced by starch with I2.
2. Take 3 ml of glycogen solution and add 6 ml of 95% alcohol. Note the precipitation of
glycogen.
3. Test the precipitate obtained from 2 with Fehling’s solution, boiling only for a short time. Note
the result.
4. Hydrolyze 5 ml of the glycogen solution with a few drops of concentrated HCl, boiling for 10
minutes. Cool solution, neutralize with NaOH and test with Fehling’s solution. Explain the
result obtained.
RESULTS AND DISCUSSIONS:
Answer:
This experiment was divided into two part; the first part is the preparation of the glycogen
sample and the second part is the reaction test of the isolated glycogen sample. The
preparation consist of the grinding of the meat of the seashells and sand giving us an animal
starch. After adding 40 ml of water it was boiled for 20 minutes giving us a light to pinkish
color as a result. After boiling it was then filtered and added with acetic acid, with the use of
blue litmus paper we tested its acidity; when it turns red it could be said that the solution is
already acidic. After acidifying the solution it was filtered again giving us an isolated glycogen
sample, it has a milky white color. Then the isolated glycogen sample was added with iodine
solution and giving as a mixture with a reddish brown color. So after adding iodine solution to
the isolated glycogen sample we then compare the result from the previous result in the
 experiment in carbohydrates specifically the starch, upon comparison and observation I could
say that they both have different colors, the color of the starch from the experiment in
carbohydrates is blue-black color and the resulting color of glycogen with iodine solution is
reddish brown. So after comparing we then go to the next experiment which was the addition
of 6 ml of 95% alcohol into the tube with an isolated glycogen sample and the result is that
the glycogen is insoluble in alcohol which then creates a precipitation that traps the nucleic
acid then a visible pellet is formed, which greatly facilitates handling of the precipitated nucleic
acids. Then the result of the precipitated alcohol and glycogen was added with Fehling’s
solution and was the boiled and after boiling it, upon observation the precipitated solution with
Fehling’s solution was mixed into one and it has become soluble and color of the mixture is
blue. For the experiment number 4 the mixture that we obtained has a brownish red color,
this was due to the heat reducing the sugars in the mixture and this also helps in the detection
of reducing sugars in the test solutions.
APPLICATION:
1. What is the difference between starch and glycogen?
Answer: The difference between starch and glycogen; starch is produced by vegetables, it is
made up of two molecule, and has both chain and branched components while glycogen is
produced by animals and it is known as animal starch, it is only made up of one molecule,
and has a branch structure. The color between the two is different, the color starch is blue-
black while for glycogen the color is reddish brown.
2. What is the purpose of hydrolyzing the glycogen before submitting it to Fehling’s test?
Answer: the purpose of hydrolyzing the glycogen before going to Fehling’s test so that the
glycogen can be broken down into its D-glucose subunits and it would become soluble.
REFERENCES:
Tebb, M.C. (1898) hydrolysis of glycogen. Retrieved from:
https://physoc.onlinelibrary.wiley.com/doi/pdf/10.1113/jphysiol.1898.sp000704
Brust, H., Orzeckowski, S., Fettke, J (2020) Starch and Glycogen Analyses: Methods and
Techniques. Retrieved from:
https://www.researchgate.net/publication/342855958_Starch_and_Glycogen_Analyses_Met
hods_and_Techniques
Galewska, Z., Gogiel, T., Malkowski, A., Romanowicz L., Sobolewski, K., Wolañska, M. (2013)
Glycogen synthesis and degradation. Retrieved from:
https://www.umb.edu.pl/photo/pliki/WL_jednostki/zaklad_biochemii_lekarskiej/pdf/bioche
mistry_workbook.pdf
Kumar, D., Bhat, A. (2017) Glycogenesis. Retrieved from:
https://www.researchgate.net/publication/323691879_Glycogenesis
Costales, A. (2015) exercise 7 isolation of glycogen, exercise 8 determination of glycogen
purity. Retrieved from:
https://www.academia.edu/14939091/Glycogen_isolation_and_determination_of_its_pur
ity