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Objective:
To study the relationship between two carbohydrates solution with saliva and
hydrochloric acid.
Apparatus:
Boiling tubes
Pipette filler
Water bath ( 37-40
Beaker
Metal test tube racks
Graduated glass pipette ( 10ml )
Pasteur pipette
5ml Measuring cylinder
Labelling paper
Materials:
Procedure:
First part
1) A few drop of solution A and B is added on a white tile separately.
2) 2 drops of iodine solution is added to mix with each solution by using a glass
rod.
3) The observation is recorded in Table 1.
Second part
1) 2ml of solution A and 2ml of solution B are added into two boiling tube each.
2) 2ml of Benedicts solution is added to each boiling tube.
3) Both boiling tube is heated in the water bath ( around 90-95
minutes.
4) The observation is record in Table 1.
) for two
5) 10ml of solution B is pipetted into each of four boiling tube. The boiling tube is
labelled with number 1, 2, 3 and 4 respectively with labelling paper near the
mouth of the test tube.
6) Boiling tube 1 and 2 is placed into water bath of ~37
process, the 4 boiling tubes is shook and the observation after one minute is
recorded in Table 2.
15) Boiling tube with label 1, 2, 3, 4, (1), (2), (3) and (4) is washed and 4 boiling
tube is labelled 1, 2, 3 and 4.
16) At the 35th minutes of incubation, boiling tube labelled 1, 2, 3 and 4 is poured
out about one-third of the total volume of the content into the newly labelled
boiling tube.
17) The acid in the boiling tube 2 and is neutralized with 2ml of sodium
hydroxide each.
18) 1ml of the solution in the boiling tube 1, 2, 3 and 4 is drawn out into new
boiling tube and labelled (1), (2), (3) and (4). 1ml of Benedicts solution for
each boiling tube is added. By using test tube holder, the newly labelled
boiling tube is heated in the water bath~95
process, the 4 boiling tube is shook and the observation after one minute is
recorded in Table 2.
Results:
Solution A
Observations
Benedicts test: brick-red solution is
formed
Iodine test: yellow solution is formed
Solution B
Temperature
Tube
Contents
10ml solution B
1ml saliva
37
10ml solution B
1ml 3M HCl
37
Conclusions
Presence of reducing sugar
No starch if presence
No reducing sugar is presence
Presence of starch
Blur colour is
remained.
10ml solution B
1ml HCl
95
10ml solution B
1ml 3M saliva
95
Blue colour is
remained.
Table 2
Discussion:
This experiment contained 2 parts. The first part of the experiment is to test between
solution A and solution B which one is reducing sugar. The second part of the experiment is to
test and investigate the type of reactions. There are 2 types of reaction which is the biological
reaction and chemical reaction. Hydrochloric acid is the chemical enzyme and salivary amylase
is the biological enzyme.
There is an enzyme reaction in this experiment which is hydrolysis reaction. Solution B is
polysaccharide which will be break down into monosaccharide during the hydrolysis reaction.
Hydrochloric acid is strong acid, it will corrode or digest any object that in contact with it.
Hydrochloric acid will acts as chemical enzyme can hydrolyse polysaccharide at high
temperature. For solution B and starch, hydrochloride acid will break them down into their own
respective monosaccharide.
Temperature will have effect on saliva and may have effect on solution A and solution B.
of reaction. But high temperature will cause enzyme itself undergo denaturation. The shape of
the active site of the enzyme changes and causing from the bond between the enzymes itself
disturbed as heat energy is gained. As the shape changes the enzyme cannot function as usual,
thus no reaction can be taken out then the rate of reaction will drop or even does not occur. For
the case of solution B, at high temperature content in solution B gains heat thus increase the
kinetic energy, the activation energy can be achieve easily then it is easier to be catalysed by
the hydrochloric acid.
Products formed after the hydrolysis reaction is glucose as its been broken into
monosaccharide. This is because glucose is reducing sugar that will reduce the copper ions (
2+
+
precipitate in the solution. Thus as we can see in the observation as brick-red precipitate is
observed.
the maximum reaction. In the other hand, the hydrochloric acid cannot carry out reaction in this
temperature as the temperature is not high enough to get that energy to catalyse the reaction.
But when in high temperature, hydrochloric acid will able to convert or break down starch to
glucose molecule because it gains enough energy from high temperature. But, salivary amylase
will denature at this high temperature. This condition will disturb the bond that hold the threedimension shape of the enzyme that altered the shape of the enzyme therefore the starch
molecule cannot bind with the denatured enzyme or active site altered enzyme. Benedicts
solution will not be success when in acidic condition. Therefore the alkaline is added before
running the Benedict test. The alkaline used is sodium hydroxide to neutralize the acidic
condition. Then the Benedicts solution is added to test the presence of reducing sugar for
example glucose.
Carbohydrate Solution B is the complex molecule so it will no show the positive result
of the reaction inBenedicts test. Before hydrochloric acid is added, solution B still is starch.
After hydrolysing reaction take place, is shows positive result to the Benedicts test that means
glucose is present in solution B after hydrolysing.
Conclusion:
Hydrolysis reaction involve salivary amylase enzyme can only take place when it is in
the optimum temperature condition ~37 . But will not show significant positive result in
high temperature condition~95 . Therefore, it will only show positive result in ~37
temperature condition. Hydrochloric acid required high temperature ~95
in order to
catalyse the reaction so it will only show positive result in the high temperature~95
condition. Both the result as shown in the Table 1 and Table 2 is same with the theory.
References:
Phil Bunyan. 2007. Two hydrolytic enzymes and an epistemologicalhistorical approach.
http://www.scienceinschool.org/2007/issue4/enzymes
http://brilliantbiologystudent.weebly.com/benedicts-test-for-reducing-sugars.html