Hydrolysis of Starch

Hydrolysis of Starch
Background:
Polysaccharides, such as amylase or amylopectin, are polymers of
glucose molecules. Starch can form an intense, brilliant, dark blue or
violet colored complex with iodine. The straight chain component of
starch (or amylose) gives a blue color, while the branched component
(or amylopectin) yields a purple color. In the presence of iodine,
amylose forms helixes, where the iodine molecules assemble as long
polyiodide chains. The helix– forming branches of amylopectin are
much shorter than those of amylose. Therefore, the polyiodide chains
are also much shorter in the amylopectin–iodine complex than in the
amylose–iodine complex. The result is a different color (purple). When
starch is hydrolyzed and broken down to small carbohydrate units, the
iodine will not give a dark blue (or purple) color. The iodine test is used
in this experiment to indicate the completion of the hydrolysis.
Amylase is an enzyme found in the saliva as well as the intestines
of humans and animals. It begins the breakdown of starch molecules in
the mouth as food is chewed and finishes its job in the intestines.
Since it is a protein, it will coagulate and be rendered useless if it is
treated with high heat.
Purpose:
In this lab two different methods will be applied to break down starch
molecules into smaller units containing of 2 to 3 glucose molecules. In
the first part of the experiment acid and heat will be used to hydrolyze
the starch chains. In the second part, and enzyme will be used to
break apart the starch chains.
Materials:
starch solution, 10% iodine solution, 2M hydrochloric acid, 1M sodium
hydroxide, amylase, Fehling’s solution A, Fehling’s solution B, 3 test
tubes, test tube rack, hot water bath, pipettes, 24-well plate, small
beaker with rinsing water, small waste water beaker
Safety:
Wear goggles and gloves. Be aware that hot plates stay hot for a long
time after they are turned off.
Procedure:
Hydrolysis of Starch 1
Hydrolysis of Starch with Acid:
1. Start the hot water bath.
2. Place a few drops of the starch solution into a well on the spot
plate. Add a couple of drops of the 10% iodine solution and record
the color.
3. Heat the test tube with the starch solution in the water bath for 1
minute and repeat step 2.
4. Add 1.0 mL of dilute hydrochloric acid to the test tube and heat
the solution.
5. After two minutes of heating perform the iodine test for starch
(step 2).
6. Keep heating the solution and test every two minutes for starch
until the solution no longer tests positive for starch. Make sure to
rinse the pipette with distilled water after each test.
7. When the solution no longer tests positive for starch, add 10
drops of NaOH to the starch solution until the solution is slightly
basic. (Place a drop of the solution onto Litmus paper. When the
paper turns blue, the solution is basic.)
8. Perform the Fehling’s test on the starch solution.
Hydrolysis of Starch with and Enzyme:

1. Add a tiny amount of amylase to a test tube containing 10.0 mL of
starch solution and swirl the test tube to mix.
2. Test the solution for the presence of starch.
3. Place the solution into a warm water bath (body temperature).
4. Every two minutes test the solution for presence of starch.
Observations:
Starch solution and iodine:
Hydrolysis with acid

Time Color Time Color Time Color
2 min 12 22
min min
4 min 14 24
min min
6 min 16 26
min min
8 min 18 28
2 Hydrolysis of Starch
min min
10 20 30
min min min
Fehling’s Test:
Hydrolysis with enzyme

Time Color Time Color Time Color
2 min 12 22
min min
4 min 14 24
min min
6 min 16 26
min min
8 min 18 28
min min
10 20 30
min min min
Questions:
1. What indicates a positive result for the Fehling’s Test?
2. Does starch give a positive or a negative result for the Fehling’s

Test?
3. Draw a representation of a short section of the starch molecule.
4. Plant starch is made up of amylose and amylopectin. Draw a

sketch to show the difference between these two forms of
starch?
5. The starch stored in the liver and muscles of animals is called

glycogen. Does glycogen resemble amylose or amylopectin?
6. What is the main function of starch?
7. Write an equation for the formation of starch from glucose

molecules.
4 Hydrolysis of Starch
8. Why is the above reaction called a dehydration reaction?
9. Why is NaOH added to the solution before the Fehling’s Test is

performed?
10. Compare the hydrolysis of starch with acid to the hydrolysis

of starch with amylase.

Hydrolysis of Starch

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Hydrolysis of Starch

Hydrolysis of Starch with and Enzyme:

Hydrolysis with acid

Hydrolysis with enzyme

2. Does starch give a positive or a negative result for the Fehling’s

4. Plant starch is made up of amylose and amylopectin. Draw a

5. The starch stored in the liver and muscles of animals is called

6. What is the main function of starch?

7. Write an equation for the formation of starch from glucose

9. Why is NaOH added to the solution before the Fehling’s Test is

10. Compare the hydrolysis of starch with acid to the hydrolysis

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