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CARBOHYDRATES

Carbohydrates are important in our daily energy requirement. They are found in both
plants and animals where they perform both structural and metabolic roles. In plants, glucose,
a very important monosaccharide, is synthesized from carbon dioxide and water using solar
energy through the process photosynthesis. It is stored as starch or converted to cellulose
which forms the plant's structural framework. Although man can synthesize some forms of
carbohydrates from fats and proteins, his main source of carbohydrate intake are plants. The
knowledge of the properties, structures and physiologic significance of carbohydrates is very
important in understanding their role in the energy use of organisms.

An important carbohydrate absorbed and metabolized in the liver is glucose. Glucose is

the simplest carbohydrate from which other polysaccharides are derived and is considered to
be the major fuel in all living animal cells.

Carbohydrates are aldehyde or ketone derivatives of polyhydric alcohols. The following

are the classifications of carbohydrates:

a. Monosaccharides - carbohydrates that cannot be hydrolyzed further

into simpler forms. Examples are erythrose, ribose and glucose.
b. Disaccharides - carbohydrates that yield two molecules of similar or
different monosaccharides upon hydrolysis. Examples are sucrose,
lactose and maltose.
c. Polysaccharides - are carbohydrates that yield 6 or more molecules of
similar or different monosaccharides upon hydrolysis. Some examples
are amylose, glycogen and starch.

A number of reactions are used in the characterization of carbohydrates. They are

divided primarily into

a. Fragmentation of carbohydrates by strong mineral acids - Molisch

α-naphthol reaction, Bial's orcinol test and Seliwanoff test
b. Reduction of metallic ions by sugars - Benedict's, Barfoed's, Tollen's
and Fehling's Tests
c. Oxidation of Carbohydrates - Phenylhydrazine reaction and Mucic acid
test
d. Hydrolysis of carbohydrates by dilute acids or enzymes - Iodine Test
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Experiment 8 and 9
EXTRACTION OF LIVER GLYCOGEN AND PLANT STARCH /
GENERAL TESTS FOR CARBOHYDRATES

OBJECTIVES

1. To extract glycogen from liver and starch from a plant sample.

2. To determine the presence of carbohydrates in extracts using Iodine
and Molisch Tests.

MATERIALS

1000 mL beaker (for 1 class) glass funnel

2 pcs 250 mL beaker wire gauze
electronic balance spatula
50 mL graduated cylinder test tube brush
4 test tubes test tube holder
test tube rack water bath
filter paper blender
tripod cheese grater
stirring rod cotton cloth
10 mL syringe kitchen knife
bunsen burner chopping board
medicine dropper

Reagents:
10% trichloroacetic acid solution (TCA)
95% ethanol
NaCl
Molisch Reagent
Iodine solution
concentrated sulfuric acid (H2SO4) solution

PROCEDURES

A. Extraction of Liver Glycogen

1. Process in a blender 250 grams of sliced chilled liver and 250 mL of

10% TCA solution.
2. Strain the mixture through a cotton cloth. Collect the filtrate in a 1000-mL beaker.
3. Divide the collected filtrate among the total number of groups.
Measure the volume of the extract obtained.
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4. While stirring the filtrate, slowly add 95% ethanol. (Ratio: 2 mL of

ethanol for every 1 mL of the filtrate).
5. Allow the mixture to stand for a few minutes until a clear liquid
separates from the mixture. If no separation occurs, add a pinch of
NaCl to the mixture then warm in a water bath.
6. Carefully separate the clear liquid from the mixture using a syringe. 7. Place the
liquid in two separate test tubes. Set aside for procedure C.

B. Extraction of Starch from Plant Sample

1. Grate the plant sample using the cheese grater.

2. Add 30 mL of water to the grated sample. Mix well.
3. Squeeze the liquid from the sample using a cotton cloth. Collect the
extract.
4. Allow the liquid to stand until a visible residue settles at the bottom of
the container.
5. Decant the mixture slowly until about 2 mL of the liquid remains.
6. Divide the residue into two parts and place in two separate test
tubes. Set aside for procedure C.

C. General Test for Carbohydrates

1. Iodine Test

a. Place 1 drop of iodine in each extract collected in procedures A and B.

b. Take note of the colors obtained.

2. Molisch Test

a. To 1 mL of the extracts obtained in procedures A and B, add 2 drops of

Molisch reagent. Mix thoroughly.
b. Incline the test tubes and slowly allow 2 mL of concentrated H2SO4 to flow
down the wall of the tube.
c. Take note of the color formed at the junction of the two liquids.
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REPORT SHEET

Group No.: _______________________________ Score: _______________

Name: __________________________________ Date Performed: _______
C/Y/S: ___________________________________ Date Submitted: _______

Experiment 7 and 8
EXTRACTION OF LIVER GLYCOGEN AND PLANT STARCH /
GENERAL TESTS FOR CARBOHYDRATES

DATA and RESULTS

Color Obtained
Extract Iodine Test Molisch Test
Glycogen
Starch

POST LABORATORY DISCUSSION

1. Describe the appearance of the extracts from the liver and plant samples.
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2. What is the purpose of adding TCA to the liver sample? Why is there a need to lower the
temperature of the mixture after the addition of TCA?
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3. Differentiate the extraction processes used for glycogen and starch.

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4. Is there any difference in the colors produced by glycogen and starch in the Iodine Test?
Describe the difference.
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5. What is Molisch Test? Explain/Describe the reaction mechanism behind the test.
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CONCLUSION

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CLINICAL APPLICATIONS

1. What is the importance of carbohydrate in the body? Enumerate some natural sources of
carbohydrates that are found in the diet.
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2. Name some diseases associated with carbohydrate deficiency.

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3. Is there a relationship between mental retardation and chronic/long-term carbohydrate-

deficient diet? Explain.
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