EXPEIMENT 4 : DIGESTION OF STARCH IN HUMANS

OBJECTIVE

1. To inspect the hypothesis that saliva contain an enzyme which break down starch to
sugar.
2. To investigate the properties of the enzyme and the conditions under which it works most
effectively.

MATERIALS

1. Test tube (12 / group)

2. Test tube rack
3. Measuring cylinder (20 cm3)
4. Water bath
5. Starch suspension
6. Dextrose (glucose)
7. Iodine solution
8. Benedict’s reagent
9. Hydrochloric acid (1 mol dm-3)
10. Aluminium foil

PROCEDURE

1. 20 cm3 of uncontaminated saliva was collected in a measuring cylinder.

2. If you cannot produce 20 cm3 of saliva, make do with 10 cm3 and all subsequent
quantities and reagents was divided.
3. One quarter of the saliva was transferred to a test tube and placed this in a boiling
water bath for 15 minutes.
4. A further quarter was transferred to another test tube and with 2 drops of hydrochloric
acid (1 mol dm-3) was added and mixed well and left for at least 15 minutes. The rest
of the saliva was kept at room temperature.
5. Six pairs of test tubes was set up as follows, each test tube was labelled with code
number as in Table 1.
 Label Content
Pair A 1 2 cm3 sugar only
2 2 cm3 sugar only
Pair B 3 2 cm3 starch only
4 2 cm3 starch only
Pair C 5 2 cm3 untreated saliva only
6 2 cm3 untreated saliva only
Pair D 7 2 cm3 starch plus
1 cm3 untreated saliva
8 2 cm3 starch only
1 cm3 untreated saliva
Pair E 9 2 cm3 starch plus
1 cm3 pre-heated saliva
10 2 cm3 starch plus
1 cm3 pre-heated saliva
Pair F 11 2 cm3 starch plus
1 cm3 acidified saliva
12 2 cm3 starch plus
1 cm3 acidified saliva
Table 1 : Test Tube Setup

6. The test tubes was left at least 10 minutes before proceeding further.
7. The content of each pair of test tube was tested :
 For starch, by adding 1 drops of iodine solutions.
 For sugar, by heating with 1 mL of Benedict’s reagent.
8. The starch test was used on the first of each pair of test tube, and the sugar test on
the second of each pair.
9. To facilitate comparison, the test tubes in the rack was arranged in numerical order.
10. The colour changes was recorded in a Table 2. Indicate which test tubes show positive
results and which ones show negative results for each starch and sugar contents.
RESULTS

Test Tube Label Content Observation Conclusion

Pair A 1 2 cm3 sugar only Yellow Starch absence

2 2 cm3 sugar only Red brick Sugar presence
Pair B 3 2 cm3 starch only Dark blue Starch presence
4 2 cm3 starch only Greenish blue Sugar absence
Pair C 5 2 cm3 untreated saliva only Cloudy yellow Starch absence
6 2 cm3 untreated saliva only Cloudy blue Sugar absence
Pair D 7 2 cm3 starch plus Partially yellow Starch is partially
and dark blue digested
1 cm3 untreated saliva
8 2 cm3 starch plus Orange Sugar is partially
digested
1 cm3 untreated saliva
Pair E 9 2 cm3 starch plus Dark blue and Starch presence
cloudy layer
1 cm3 pre-heated saliva
10 2 cm3 starch plus Light blue Sugar absence
1 cm3 pre-heated saliva
Pair F 11 2 cm3 starch plus Dark blue Starch presence
1 cm3 acidified saliva
12 2 cm3 starch plus Light blue Sugar absence
1 cm3 acidified saliva
Table 2 : Enzyme Reactions For Sugar and Starch
DISCUSSION

Human’s saliva, there are many substances that had been consumed by us everyday
such as protein carbohydrates, fibres and much more. Some of it contains starch. Starch is a
polysaccharide which has to be broken down into free sugar molecules before absorption.
Starch is mixed well with saliva under various conditions. After a period of time, the mixture
is tested with iodine solution for starch while for sugar with Benedict’s reagent. The role of
amylase in food digestion by using breakdown of starch from polysaccharide to
monosaccharide in reducing the sugar form.

The experiment was carried out on the human saliva to investigate the action of saliva
and 3 M Hydrochloric Acid in two carbohydrate solutions. The salivary amylase is also known
as ptyalin is involved in this experiment which is produced by salivary gland in mouth. The
complex starch broke down into simple sugars by the help of the salivary amylase, so that the
sugar can be easily converted into energy. The enzyme does not breakdown carbohydrates
to its simplest form. The enzyme also acted as the biological catalyst which helps to reduce
the activation energy needed for the Benedict’s test. When the temperature increases to 95C,
the enzyme denature the increasing of temperature cause the molecules to vibrate more
violently. The weak bonds of the enzyme such as hydrogen bond will break first. The enzyme
bond of enzymes such as ionic bonds and disulphide bridges break later. Structure and surface
of active site are destroyed and altered. The catalytic function of enzyme losses and it said to
be denatured. When the structure of the enzyme is fully destroyed, they will be no catalysis
take place on it. Hydrochloric acid, HCl hydrolyse starch into glucose at temperature 95C.
Therefore, glucose was present in tube 2. Tube 2 has incubated at optimum temperature of
HCl, thus HCl hydrolyse starch (Solution B) into glucose which show the positive result also
tested with Benedict’s solution. The blue solution turns to yellowish brown and lastly the
solution forms brick red precipitation. This is because with the temperature reach 95C,
hydrolysis still occurs before it undergoes denaturation. To prevent this, enzyme can be heated
until it reached 95C. Then use it to react with the solution B instead of heating them together
by the presence of starch and sugar can be tested by adding iodine solution and Benedict’s
reagent into the specimen which is the saliva or others specimen. For test tubes Pair A contains
sugar only while for Pair B contains starch only. When the iodine solution and the Benedict’s
reagent added into it, the colour of the solution for Pair A which had been added with
Benedict’s reagent will change colour to red brick while for Pair B which had been added with
iodine solution will change colour to dark blue. For this experiment, our saliva can be tested
by adding these two reagent investigate whether there are any sugar which had been tested
by adding Benedict’s reagent or any starch which had been tested by adding iodine solution.

For test tube 5, 6, 7, and 8, the saliva which had been tested contains an enzyme
which breaks down the starch to sugar. The enzyme that was used to break down the starch
to sugar is the amylase and glucose. For the test tube 5, 6, 7, and 8 the saliva that was used
for this test tubes is the untreated saliva which the saliva does not undergo any process which
can make the enzyme of the saliva to denatured. That is why the enzyme still remain in the
saliva and used to break down the starch to sugar. For test tube 5 and 6, the colour does
change but not in the colour which was the starch and sugar presence. For 7 and 8, the
present of the starch and sugar can be found because the starch and sugar was partially
digested which make the colour still changing but partially.
POST LAB QUESTION

1. Why must the contents of test tubes be left undisturbed for at least 10 minutes before
starting the analysis?
 The contents of test tubes have be left undisturbed for at least 10 minutes before
starting the analysis is to let the enzyme reacts completely with the solution.
2. What is the purpose of testing test tube 1 to 6?
 The purpose of testing test tube 1 till 6 were to control the experiment or variable.
3. Based on your experiment, which of the test tube are used to test the hypothesis that
saliva contains an enzyme which breaks down starch to sugar?
 The test tube are used in the test of the hypothesis that saliva contains an enzyme
which breaks down starch to sugar are test tube 5, 6, 7, and 8.
4. What is the name of the enzyme in saliva and what composed does it produce from
starch?
 The enzyme in saliva and, that composed does it produce from starch is amylase or
glucose.
5. Did the hypothesis turn out to be correct? Would you say it was proven?
 Yes, it was proven because the untreated saliva do not undergo any condition that
can denatured the enzymes.
6. Which test tube provides information on the properties of the enzyme?
 Test tubes 5, 6, 7, and 8 are provides information on the properties of the enzyme.

CONCLUSION

In this experiment, the hypothesis that saliva contain an enzyme which break down
starch to sugar was inspected which can be found in the test tube 5, 6, 7, and 8. The properties
of the enzyme and the conditions under which it works most effectively was investigated and
it is also can be found in the test tubes 5, 6, 7, and 8.
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Neil, A. C., & Jane, B. R. (2009). Campbell Biology - 9th edition. San Francisco : Pearson
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Reece, J. B., & Campbell, N. A. (2011). Campbell Biology - Tenth Edition . USA : Pearson
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Rusell, P. J., Hertz, P. E., & McMillan, B. (2014). Biology : The Dynamic Science - Fourth
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Taylor, D. J., Green, N. P., Stout, G. W., & Soper, R. (1997). Biological Science ! & 2 .
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