CHAPTER 4:

Experiment 4.1
Aim

To study the effects of temperature on the activity of enzyme amylase on starch.

Problem statement:
What is the effect of temperature on the activity of amylase on starch?
Variables

a) Manipulated variable : Temperature of reaction medium

b) Responding variable : Time taken for the complete hydrolysis of starch
c) Constant variable
Hypothesis

: Substrate concentration

The higher the temperature, the higher the rate of enzymatic reaction until it reaches
its optimum temperature of 37 C.
Materials

1% starch suspension iodine solution, ice, distilled water and 1% of enzyme amylase.
Apparatus

Five beakers, ten test tubes, syringe, dropper, glass rod, white tile, thermometer,
Bunsen burner, tripod stand, wire gauze and stopwatch.
Procedure

1. Rinse your mouth with clean water and collect 5 ml of saliva in a beaker.
2. Add 5 ml of distilled water into the beaker to dilute the saliva.
3. Label five tubes as A, B, C, D, E and fill up each with 1ml of saliva using a
syringe.
4. Label five test tubes as A1 , B1 , C1 , D1 , E1 and fill each with 5ml of 1% starch
suspension using a second syringe.
5. Place test tubes A and A1 into a beaker of cold water and maintain it at
temperature of 5 C. Leave it for 10 minutes.
6. Drop few drops of iodine separately on a while tile.
7. After 10 minutes, pour the content in test tube A into test tube A1 . Maintain
the temperature of the mixture at 5 C.
8. Stir the mixture with a glass rod. Use a dropper to take out small amounts of
the mixture and add it to the iodine drop. Start the stopwatch immediately.

9. Observe the change in the colour of the iodine.

10. Repeat the iodine test at an interval of 1 minute for 10 minutes.
11. Record the title when the mixture no longer changes the colour of iodine.
12. Repeat steps 5 to 11 with the pair of test tubes at different temperature as
shown in the table below.
Pair of test tubes
Temperature ( C)

B, B1
28

C, C1
37

D, D1
45

E ,E1
60

13. Record all your observations.

14. Plot a graph of rate of reaction

Results

1
]
time against temperature ( C) .

Temperature( C)
5
28
37
45
60

Time taken for the

complete hydrolysis of
starch (minute)

Rate of reaction
(min-1)

1
]
time

Experiment 4.2
Aim

To study the effects of pH on the activity of pepsin.

Problem statement:
What is the effects of pH on the activity of pepsin?
Variables

a) Manipulated variable

: pH of solution

b) Responding variable

: Time taken for content in the test tube to turn

clear
c) Constant variable
Hypothesis

: Temperature of solution

Pepsin works best in an acidic medium at pH 3.

Materials

Albumen suspension, 1% pepsin solution, 0.1 M hydrochloric acid, 0.1 M sodium

hydroxide solution, pH paper and distilled water.
Apparatus

Test tubes, syringe, thermometer, stopwatch, beaker, Bunsen burner, tripod stand,
wine gauze and filter funnel.
Procedure

1. Pour 5 ml of albumen suspension into each of the three test tubes, labelled as P,
Q and R.
2. Add the following solutions into each test tube according to the table below.
Test tube
P
Q
R

Solution
1ml of 0.1 M hydrochloric acid + 1ml of 1% pepsin solution
1ml of 0.1 M sodium hydroxide solution + 1ml of 1% pepsin solution
1ml distilled water + 1ml of 1% pepsin solution

3. Dip a piece of pH paper into each test tube. Record the pH value.
4. Place all test tubes into a beaker bath at 37 C.
5. Determine the time taken for the content in each test tube to turn clear using a
stopwatch. Record all data.

6. Calculate the rate of reaction using the formula

Results
Test tube

1
]
time .

:
pH

Time taken for the content in

test tube to turn clear (min)

Rate of reaction
(min-1)

P
Q
R

1
]
time

Experiment 4.3
Aim

To study the effects of substrate concentration on the activity of salivary amylase.

Problem statement:
What is the effects of substrate concentration on the activity of salivary amylase?
Variables

a) Manipulated variable

: Substrate concentration

b) Responding variable

: Time taken for hydrolysis of starch to be

completed
c) Constant variable
Hypothesis

: Temperature of solution

The higher the substrate concentration, the higher the rate of biochemical reaction
until it reaches a maximum rate.
Materials

Starch suspensions at various concentrations (0.1%, 0.2%, 0.3%, 0.4%, 0.5% and
0.6%), 0.1% saliva suspension and iodine solution.
Apparatus

Test tubes, syringes, glass rod, stopwatch, white tiles with grooves, droppers and
measuring cylinders
Procedure

1. Prepare 10 ml of 0.1% salivary amylase.

2. Prepare six test tubes labelled as A, B, C, D, E and F.
3. Pour 4 ml of starch suspensions of various concentrations into the test
tubes using different syringes as shown in the table below.
Test tube
A
B
C
D
E

Concentration of starch suspension (%)

0.1
0.2
0.3
0.4
0.5

0.6

4. Immerse the test tubes in a water bath at 37 C.

5. Add drops of iodine solution separately onto the grooves of the white tile.
6. Add 1 ml of 0.1% amylase to test tube A using a syringe.
7. Start the stopwatch immediately. Stir the contents with a glass rod. Repeat
this step at 30-second intervals until the mixture stops turning blue-black
in colour when tested with iodine solution.
8. Repeat steps 6 and 7 with test tubes B, C, D, E and F. At every sampling,
rinse the dropper with clean distilled water.
9. Record the results in a table.
10. Plot a graph of rate of reaction

Results
Test tube

1
]
time against substrate concentration.

:
Concentration of

Time taken for the

starch suspension

complete hydrolysis

(%)

of starch (minute)

A
B
C
D
E
F

Experiment 4.4

0.1
0.2
0.3
0.4
0.5
0.6

Rate of reaction
(min-1)

1
]
time

Aim

To study the effects of enzyme concentration on the activity of salivary amylase.

Problem statement:
What is the effects of enzyme concentration on the activity of salivary amylase?
Variables

a) Manipulated variable : Enzyme concentration

b) Responding variable : Time taken for hydrolysis of starch to be completed
c) Constant variable
Hypothesis

: Temperature of solution

The higher the enzyme concentration, the higher the rate of biochemical reaction until
it reaches a maximum rate.
Materials

1% starch suspension, 0.5% saliva suspension, iodine solution and distilled water.
Apparatus

Test tubes, syringes, glass rod, stopwatch, white tiles with grooves, droppers and
measuring cylinders
Procedure

:
1. Label six test tubes as A, B, C, D, E and F.
2. Fill each test tubes with different volumes of saliva and distilled water
as shown in the table below.
Test tube
A
B

A
0.5
2.5

B
1.0
2.0

C
1.5
1.5

D
2.0
1.0

E
2.5
0.5

F
3.0
0.0

3. Immerse the test tubes in a water bath at 37 C.

4. Add drops of iodine solution separately onto the grooves of the white
tile.
5. Add 4 ml of 0.5% saliva suspension to test tube A using a syringe.
6. Start the stopwatch immediately. Stir the contents with a glass rod.
7. Draw a small amount of the mixture using a dropper and add to a drop
of iodine on the white tile immediately.
8. Observe the colour change in iodine.

9. Carry out the iodine test at an interval of 30 seconds until the mixture
stops turning blue-black in colour when tested with iodine solution.
10. Record the time taken when the colour of iodine remained.
11. Repeat steps 5 to 10 for test tubes B, C, D, E and F. At every sampling,
rinse the dropper with clean distilled water.
12. Record the results in a table.
13. Plot a graph of rate of reaction

1
]
time against enzyme

concentration.

Results

Test tube

Enzyme

Time taken for the

concentaration (%)

complete hydrolysis
of starch (minute)

A
B
C
D
E
F

Experiment 6.1

Rate of reaction
(min-1)

1
]
time

Aim

To determine the vitamin C content in fruit juice.

Problem statement:
What fruit juice has higher vitamin C content?
Variables

a) Manipulated variable

: Type of fruit juice

b) Responding variable

: Volume of fruit juice needed to decolourise

DCPIP solution

c) Constant variable
Hypothesis

: Amount of DCPIP solution.

Lemon juice has a higher vitamin C content than that of pineapple and papaya juice.
Materials

0.1% ascorbic acid solution, 1.0% dichlorophenolindophenol solution (DCPIP),

lemon juice, pineapple juice and papaya juice.
Apparatus

Specimen tubes, syringers and measuring cylinder (5ml)

Procedure

1. Label four specimen tubes as A, B, C dan D.

2. Place 1 ml of DCPIP solution in each specimem tube.
3. Fill a syringe with 5 ml of ascorbic acid solution.
4. Immerse the needle of the syringe in the DCPIP solution as shown in the diagram.
5. Add the ascorbic acid solution to the DCPIP solution drop-by-drop, and shake the
tube slowly.
6. Record the volumn of ascorbic acid solution used to turn the DCPIP solution
colourless.
7. Repeat steps 3 to 6 using lemon juice, pineapple juice and papaya juice.
8. Calculate the percentage and concentration of vitamin C in these three types of
fruit juices using the formula below:

Percentage of vitamin C =

Volume of 0.1 ascorbic acid solution

0.1
Volume of fruit juice

Concentration of vitamin C =

Results
Solution

Volume of 0.1 ascorbic acid solution

0.1
Volume of fruit juice

:
Initial

Final

Volume used

volume

volume

(ml)

(ml)

(ml)

Ascorbic acid
Lemon juice
Pineapple

5.0
5.0
5.0

juice
Papaya juice

5.0

Percentag
e of
vitamin C
(%)

Concentration
of vitamin C
(mg cm-3)

Experiment 6.2
Aim

To study the digestion of starch.

Problem statement:
What is the action of salivary amylase on starch?
Variables

d) Manipulated variable : Absence or presence of starch

e) Responding variable : Presence of reducing sugar
f) Constant variable
Hypothesis

: Concentration of starch.

Salivary amylase catalyses the break down of starch to reducing sugar.

Materials

1% starch suspension, Benedicts solution, iodine solution, saliva and distilled water.
Apparatus

Test tubes, beakers, measuring cylinders (5ml), test tube holder, Bunsen burner, tripod
stand and wire gauze.
Procedure

1. Collect 2 ml of saliva in a beaker and dilute with 2 ml of distilled water.

2. Put 1 ml of diluted saliva into a test tube. Test for the presence of starch.
3. Put 1 ml of diluted saliva into another test tube. Test for the presence of
reducing sugar.
4. Repeat steps 2 and 3 using 1% starch suspension.
5. Label three test tubes as A, B and C.
6. Put 1 ml of distilled water into test tube B and 1 ml of saliva into test tubes A
and C.
7. Heat test tube C in boiling water for 5 minutes.
8. Add 5 ml of 1% starch suspension into each test tube.
9. Immerse all test tubes in a water bath of 37 C.
10. After 30 minutes, carry out iodine test and Benedicts test on the content in
test tubes A. B and C. Record all results.

14. Immerse the test tubes in a water bath at 37 C for 30 minutes.

15. Add drops of iodine solution separately onto the grooves of the white
tile.
16. Add 4 ml of 0.5% saliva suspension to test tube A using a syringe.
17. Start the stopwatch immediately. Stir the contents with a glass rod.
18. Draw a small amount of the mixture using a dropper and add to a drop
of iodine on the white tile immediately.
19. Observe the colour change in iodine.
20. Carry out the iodine test at an interval of 30 seconds until the mixture
stops turning blue-black in colour when tested with iodine solution.
21. Record the time taken when the colour of iodine remained.
22. Repeat steps 5 to 10 for test tubes B, C, D, E and F. At every sampling,
rinse the dropper with clean distilled water.
23. Record the results in a table.
24. Plot a graph of rate of reaction

1
]
time against enzyme

concentration.

Results

:
Test

Content

Iodine test

Starch suspension
Saliva
Starch suspension
Saliva

Benedicts test

Test

Test tube

Iodine test

A
B
C
A
B
C

Benedicts test

Observation

Observation

Experiment 6.3
Aim

To study the digestion of protein.

Problem statement:
What is the action of enzyme on a protein food sample?
Variables

a) Manipulated variable : Absence or presence of enzyme

b) Responding variable : Condition of solution
c) Constant variable
Hypothesis

: Volume of albumen suspension

Pepsin catalyses the hydrolysis of albumen to polypeptide.

Materials

Albumen suspension, pepsin solution, dilute hydrochloric acid and distilled water
Apparatus

10 ml pipette, 500 ml beaker, test tubes, test-tube rack, droppers, stopwatch and
thermometer.
Procedure

:
1. Label three test tubes as A, B and C.
2. Put 2 ml of albumen suspension into each test tubes.
3. Add 1 ml of pepsin solution into test tube A and B. Add 5 drops od
dilute hydrochloric acid into test tube B.
4. Put all test tubes in a water bath of 37 C.
5. Record all the observations and leave the apparatus for 30 minutes.
6. After 30 minutes, record the observations.

Results

:
Test tube

Condition of content
Beginning of experiment

A
B
C

End of experiment

Experiment 6.4
Aim

To study the effects of macronutrient deficiency in plants.

Problem statement:
What are the effects of macronutrient deficiency in plants?
Variables

a) Manipulated variable : Components of minerals in culture solution

b) Responding variable : Condition of plant
c) Constant variable
Hypothesis

: Volume of solution

: A plant is healthier if it is grown in a complete culture solution

(Knops solution).
Materials

: Maize seedlings, potassium nitrate (KNO3), potassium dihydrogen

phosphate (KH2PO4), magnesium sulphate (MgSO4), calcium nitrate (Ca (NO3)2),

ferum (III) phosphate (FePO4), distilled water, cotton wool and black paper.
Apparatus

Glass jars, rubber bungs with holes, straight glass tubes to fit into the holes of the
rubber bungs, L-shaped delivery tubes to be connected to a vacuum pump and a knife.

Procedure

1. Label eight boiling tubes as A, B, C, D, E, F, G and H

2. Fill each boiling tube with different culture solution as shown in the table below.
Boiling tube

Culture solution
Potassium
Magnesium

Ferum (III)

Distilled
water

Calcium

Potassium

nitrate

nitrate

dihydrogen

sulphate

phosphate

(0.8g)

(0.2g)

phosphate

(0.2g)

(trace)

(0.2g)
A
(Distilled water)
B
(Complete culture)
C

Replace

Replace

(No nitrogen)

with

with

calcium

potassium

chloride

chloride

Replace

Replace

(No phosphorus)

with

with ferum

potassium

(III) oxide

Replace

chloride
Replace

(No potassium)

with

with

sodium

calcium

nitrate

phosphate

Replace

(No calcium)

with sodium
nitrate

Replace with

(No magnesium)

potassium

sulphate
Replace with

(No sulphur)

magnesium
chloride

3. Place one maize seeding in each culture solution.

4. Cover all the boiling tubes with black paper.
5. Place all the boiling tubes in a condition where all the seedlings received an
equal amount of sunlight.
6. Use distilled water to top up the solution from time to time.
7. Pump air into the solution using an air pump.
8. Change the culture solution once a week.
9. Record the growth of each seeding after two weeks.

Results
Boiling tube
A
B
C
D
E
F
G
H

Experiment 6.5

:
Nutrient deficient

Effects on seedlings

Aim

To investigate the effects of light intensity on the rate of photosynthesis

Problem statement:
What is the effect of light intensity on the rate of photosynthesis?
Variables

a) Manipulated variable : Distance of Hydrilla sp. from the light source

b) Responding variable : Number of bubbles produced per minute
c) Constant variable

: Type of plant

Hypothesis

: The higher the light intensity, the higher the rate of photosynthesis.

Materials

: A few sprigs of Hydrilla sp. , 1% sodium hydrogen carbonate

solution, distilled water and plasticine.

Apparatus

: 60W bulb, 500ml beaker, a test tube, a glass funnel, a stopwatch, a

thermometer and a metre rule.

Procedure

1. Maintain the temperature of water in the beaker at 28 C

2. Choose a few strands of Hydrilla sp.
3. Make a clean oblique cut with a sharp razor near the lower end of the
Hydrilla sp. stem under water.
4. Place the aquatic plant with the bubbling end upwards, inside a glass filter
funnel. Place the funnel upside down in a beaker that contains 1% sodium
hydrogen carbonate solution.
5. Place a test tube filled with 1% sodium hydrogen carbonate solution over
the filter funnel.
6. Place a table lamp 50cm away from the beaker and switch it on.
7. After the plant releases bubbles at a constant rate, count the number of gas
bubbles released in one minute.
8. Repeat step 7 by putting the table lamp at different distances of 40cm,
30cm, 20cm and 10cm from the beaker. Record the results.
9. Plot a graph of number of bubbles produced against distance of the light
resource from the beaker
Results

Distance of the light source

(cm)
Number of gas bubbles
produced per minute

10

20

30

40

50