Results and Discussion Carbohydrates

JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY
NASC 2115 – BIOCHEMISTRY LABORATORY
IV. RESULT AND DISCUSSION
B. GLYCOGEN ISOLATION
Glycogen is a multibranched polysaccharide

of glucose that serves as a form of energy
storage in animals, fungi, and bacteria. The
polysaccharide structure represents the main
storage form of glucose in the body.
Glycogen functions as one of two

forms of long-term energy reserves, with the
Figure 1.0 and 1.1: Trituration of
other form being triglyceride stores in
Chicken liver (Top): Before Trichloroacetic
adipose tissue. In humans, glycogen is made
acid (TCA) was added; (Bottom): Triturated
and stored primarily in the cells of the liver
liver with Trichloroacetic acid
and skeletal muscle.
In the experiment, the first step was

weighing 50 grams of chicken liver then cheesecloth. The filtrate left was
placing it on a mortar. Afterwards the pestle measured in a graduated cylinder then
was used to pound the liver until it was transferred to a small beaker. 95% ethanol of
almost grounded. 50 mL of 10% twice the amount of the filtrate was added in
Trichloroacetic acid or TCA was added into the beaker. Glycogen easily separates with
the liver to triturate it completely. alcohol since polysaccharides are less soluble
than sugar in aqueous alcohol. A pinch of salt
was also added in the mixture. After letting
If the mixture was almost liquid, it was
the mixture sit for about 10 minutes, there
filtered through a
was an apparent formation of creamy white
precipitate at the bottom of the beaker. It was
made sure that the mixture was not disturbed using a dropper. What was left of the mixture
during the time it was sitting on the table. was filtered through a filter paper.
Figure 3.0: The accumulated isolated

Glycogen
After a while the leftover residue was

stuck dried on the filter paper, it was washed
off the paper using a calculated amount of
distilled water to make a glycogen solution.
Figure 2.0 and 2.1: (Top): Filtration process Glycogen is a readily mobilized
of triturated liver through the use of storage form of glucose. It is a very large,
Cheesecloth; (Bottom): Collected filtrate of branched polymer of glucose residues that
pulverized chicken liver with TCA can be broken down to yield glucose
molecules when energy is needed.
C. IODINE TEST
A warm water bath was used to help hasten
the formation of precipitate. While in the An iodine and Barfoed’s test were
conducted to a galactose, fructose, glycogen,
bath, more white precipitate can be seen paper pulp mixture (5g of paper macerated in
forming. After carefully removing the beaker 100mL water) and a starch solution
(consisting of 1 gram of corn starch boiled in
from the bath, the clearer upper layer of the 100ml of water).
mixture was removed
For the iodine test, two drops of 1% is formed and will indicate the presence of
iodine were dropped to the carbohydrates reducing sugar. The reaction will be negative
samples, and then heated over a flame, after in the presence of disaccharide sugars
a while, the galactose and fructose produced because they are weaker reducing agents.
a light yellow color, glycogen with a slight This test is specific for monosaccharides.
blue color, and paper pulp and starch Due to the weakly acidic nature of Barfoed's
produced an indigo color. reagent, it is reduced only by
monosaccharides.
A blue-black color results if starch is
present. If starch amylose is not present, then A rusty or brownish-red color
the color will stay orange or yellow. indicates monosaccharides, no color change
or weak change indicates di- and
Starch is a carbohydrate found in polysaccharides. Galactose and fructose are
plants. It consists of two different types of monosaccharides and the rest are not.
polysaccharides that are made up of glucose
units which are connected in two different
ways. One is the linear amylose and the other
is the branched amylopectin. Amylose is the Carbohydrates Iodine Barfoed’s
compound that is responsible for the blue solution reagent
color. Its chain forms a helix shape, and (Yellow- (Blue-
iodine can be bound inside this helix. Violet) Brick red)
It is thought that starch and glycogen Fructose No reaction Brick red

form helical coils. Iodine atoms can then fit (bottom)
into the helices to form a starch-iodine or
Galactose No reaction Brick red
glycogen-iodine complex. Starch in the form
(bottom)
of amylose and amylopectin has fewer
branches than glycogen. This means that the Maltose No reaction Brick red
helices of starch are longer than glycogen, (bottom)
therefore binding more iodine atoms. The Paper pulp Violet No reaction
result is that the color produced by a starch-
iodine complex is more intense than that Starch Violet No reaction
obtained with a glycogen-iodine complex.
Glycogen Violet spot No reaction
For the Barfoed’s test, test tubes
containing the carbohydrates samples were Table 2.0 This table shows the color change
added with 2.5 mL of Barfoed’s reagent and of carbohydrates in reaction to iodine
heated in a boiling water bath for 5 minutes. solution and Barfoed’s reagent.
After 5 minutes the carbohydrates samples
galactose and fructose produced a brick red
color at the bottom, glycogen yielded a blue Figure 2.0 The picture shows the result of
hue. glucose and fructose mixed and heated with
Barfoed’s reagent.
Barfoed's test is used to detect the
presence of monosaccharide (reducing)
sugars in solution. Barfoed's reagent, a
mixture of ethanoic (acetic) acid and copper
(II) acetate, is combined with the test solution
and boiled. A red copper (II) oxide precipitate
Figure 2.1 The picture shows the result of

glycogen mixed and heated with Barfoed’s
reagent.
Figure 2.1 The picture shows the result of

starch, paper pulp, fructose and galactose
mixed and heated with Barfoed’s reagent.
https://en.wikipedia.org/wiki/Glycogen
https://www.ncbi.nlm.nih.gov/books/NBK2119
0/

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JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY

NASC 2115 – BIOCHEMISTRY LABORATORY

IV. RESULT AND DISCUSSION

Glycogen is a multibranched polysaccharide

Glycogen functions as one of two

In the experiment, the first step was

placing it on a mortar. Afterwards the pestle measured in a graduated cylinder then

Figure 3.0: The accumulated isolated

After a while the leftover residue was

Cheesecloth; (Bottom): Collected filtrate of branched polymer of glucose residues that

It is thought that starch and glycogen Fructose No reaction Brick red

Figure 2.1 The picture shows the result of

Figure 2.1 The picture shows the result of

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