S.

6 Biology PHYSIOLOGICAL EXPERIMENTS

Page 1

AIM:
To determine the vitamin C content in the edible portion of lemon &
green lantern pepper.

1. You are provided with the apparatus below, design an experiment to

achieve the aim.
2. Write a logical discussion on different aspects of the experiment and make
a concise conclusion.

MATERIALS & APPARATUS:

Item
Amount
For each student on his bench
1. Towel 1 pc
2. Marker pen 1
pc
3. Razor blade 1
pc
4. White tile . . . . . . . . . . . . . . . . 1 pc
5. Mortar & pestle 1
set
6. Filter funnel 1
pc
7. Filter paper 2
pcs
8. Test-tube 6 pcs
9. Boiling tube for collecting DCPIP solution, standard vitamin C solution &
distilled
water separately . . . . . . . . . . . . . . . 3
pcs
10. Metallic test-tube rack
1 pc
11. 10-ml measuring cylinder
1 pc
12. 250-ml beaker for discarding waste
1 pc
13. 5-ml pipette 1
pc
14. Pipette filler . . . . . . . . . . . . . . . . 1
pc
15. Teat pipette 1
pc
On each students’ bench (shared by 2 students):
16. Lemon 1 pc
17. Green lantern pepper 1
pc
18. Electronic Balance 1 pc
On teacher’s bench (amount shown per student):
19. 30 ml DCPIP solution
20.  ml standard vitamin C solution (of known concentration)

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21. Weighing paper 2

pcs
On side bench (to be shared among the class):
22. Distilled water
End of experiment

The report should include:

23. the aim of the experiment
24. procedure and precaution
25. result
26. discussion (errors, suggestions for improvement etc.)
27. conclusion

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S.6 Biology PHYSIOLOGICAL EXPERIMENTS
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AIM:
To determine the vitamin C content in the edible portion of lemon &
green lantern pepper.
Method and Procedures:
1. Extract 10 ml diluted juice from a known mass of edible portion of lemon
tissue & green lantern pepper tissue separately. (the suggested size of
tissue to use is 1 cm3)
2. Collect 5 ml standard vitamin C solution.
3. Find out & record the number of drops of standard vitamin C solution
needed to decolourize a fixed volume of blue DCPIP solution.
4. Repeat steps 3 using lemon extract instead of standard vitamin C solution.
5. Repeat steps 3 using green lantern pepper extract instead of standard
vitamin C solution.
6. Calculate the concentration of vitamin C in the lemon & green lantern
pepper extract. (stating the formula used)
7. Calculate the vitamin C content in the two types of food. (stating the
formula used)
Hints:If the concentration of vitamin C in the food extract is too high (in
case the DCPIP solution is decolourized too quickly), the extract can
be further diluted to a suitable concentration & the food test be
repeated.
RESULTS:
1. List out all the raw data necessary for calculation.
2. Calculate the vitamin C content in the two types of food. (indicating clearly
the units used)
DISCUSSION:
Write a logical discussion on different aspects of the experiment and make a
concise conclusion.
MATERIALS & APPARATUS:
Item
Amount
For each student on his bench
1. Towel 1 pc
2. Marker pen 1
pc
3. Razor blade 1
pc
4. White tile . . . . . . . . . . . . . . . . 1 pc
5. Mortar & pestle 1
set
6. Filter funnel 1
pc
7. Filter paper 2
pcs
8. Test-tube 6 pcs
9. Boiling tube for collecting DCPIP solution, standard vitamin C solution &
distilled
water separately . . . . . . . . . . . . . . . 3
pcs
10. Metallic test-tube rack
1 pc
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11. 10-ml measuring cylinder

1 pc
12. 250-ml beaker for discarding waste
1 pc
13. 5-ml pipette 1
pc
14. Pipette filler . . . . . . . . . . . . . . . . 1
pc
15. Teat pipette 1
pc
On each students’ bench (shared by 2 students):
16. Lemon 1 pc
17. Green lantern pepper 1
pc
18. Electronic Balance 1 pc
On teacher’s bench (amount shown per student):
19. 30 ml DCPIP solution
20.  ml standard vitamin C solution (of known concentration)
21. Weighing paper 2
pcs
On side bench (to be shared among the class):
22. Distilled water
End of handout
TITLE:
Application of food tests (Quantitative analysis)

INTRODUCTION:
The technique of determining the concentration of food substances in an
unknown solution can be extended to area of living tissues. Before a food test
can be carried out, it is necessary to extract juice from the tissue under
investigation. Extraction technique is very common in biological
investigations.

In this practical session, you have a chance to practise how to extract juice
properly from living tissue. With the help of techniques learned in previous
practical sessions, you can roughly find out the vitamin C content (in terms of
milligram vitamin C per gram of tissue) present in two types of food.

Vitamin C (ascorbic acid) is required for the formation of intercellular material.

Deficiency of vitamin C leads to scurvy in human. Vitamin C is a powerful
reducing agent which can reduce DCPIP (2,6-dichlorophenolindophenol) &
changes the colour of DCPIP solution from blue to colourless. More vitamin C
can decolourize more DCPIP.

To determine the concentration of vitamin C in an unknown sample solution.

The amount (A ml) of standard vitamin C solution (concentration, B mg/ml)
needed to decolourize a fixed volume (C ml) of DCPIP solution is found out first.
Then the amount (D ml) of unknown sample required to decolourize the same
amount (C ml) of DCPIP solution is also found out. The concentration of

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vitamin C present in the unknown sample can be calculated by the following

formula:

Concentration of vitamin C AxB

in unknown sample = ----------------- mg/ml
D
AIM:
To determine the vitamin C content in the edible portion of lemon & green
lantern pepper.

METHOD:
1. Extract juice from a known mass of edible portion of lemon tissue according
to the following procedure:
a. Cut out a small cube (roughly 1 cm 3) of lemon tissue using a razor
blade.
b. Weigh the tissue by means of balance.
c. Grind the tissue by means of mortar & pestle to obtain as much juice as
possible.
d. Filter the juice using filter funnel & filter paper & collect the filtrate into
a 10-ml measuring cylinder.
e. Add distilled water to the macerated tissue to collect a total of 10 ml
filtrate.
f. Transfer the filtrate to a test-tube & label the tube as lemon”.
2. Repeat step 1 using the edible portion of green lantern pepper instead of
lemon. Cut the tissue cube into smaller pieces before grinding. Label the
tube containing the filtrate as pepper”.
3. Collect about 5 ml standard vitamin C solution in a test-tube labelled as
standard”.
4. Transfer 5 ml of DCPIP solution into a clean test-tube using a 5-ml pipette.
5. Add standard vitamin C solution into the 5 ml DCPIP solution drop by drop
using a teat pipette. Shake to mix the content after adding each drop.
6. Find out & record the number of drops needed to decolourize the blue
DCPIP solution.
7. Repeat steps 4 to 6 using lemon extract instead of standard vitamin C
solution.
8. Repeat steps 4 to 6 using green lantern pepper extract instead of standard
vitamin C solution.
9. Calculate the concentration of vitamin C in the lemon & green lantern
pepper extract according to the following formula:
AxB
Concentration of vitamin C in extract = ------------ mg/ml
D
(where A = number of drops of standard vitamin C solution used,
B = concentration, in mg/ml, of vitamin C solution used,
D = number of drops of extract used)
10. Calculate the vitamin C content in the two types of food according to the
following formula:
concentration of extract (mg/ml) x 10 ml
Vitamin C content = ---------------------------------------------------------
mass of tissue used (g)
RESULTS:
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Amount of standard vitamin C solution used (A) = drops

Concentration of vitamin C solution used (B) = mg/ml

Mass of lemon tissue used (M1) = g

Amount of lemon extract used (D1) = drops

A x B x 10
Vitamin C content in lemon = ------------------- mg vitamin C per g tissue
D1 x M1

= mg vitamin C per g tissue

Mass of green lantern pepper tissue used (M2) = g

Amount of green lantern pepper extract used (D2) = drops

Vitamin C content A x B x 10
in green lantern = ---------------------- mg vitamin C per g tissue
pepper D2 x M2

= mg vitamin C per g tissue

DISCUSSION:
1. What is the mechanism of DCPIP test?
2. Why is different amount of lemon & lantern pepper extract needed in
decolourizing the same amount of DCPIP solution?
3. Which food has a higher vitamin C content? What is the significance of this
finding?
4. Is it essential to measure all volumes very accurately?
5. What are the sources of error?
6. What is the limitation of this method?
7. Is there any suggestion to improve the method?

MATERIALS & APPARATUS:

Item
Amount
1. White trough 1
pc
2. Towel 1 pc
3. Marker pen 1
pc

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4. Razor blade 1
pc
5. White tile . . . . . . . . . . . . . . . . 1 pc
6. Mortar & pestle 1
set
7. Filter funnel 1
pc
8. Filter paper 2
pcs
9. Test-tube 6 pcs
10. Boiling tube for collecting DCPIP solution, standard vitamin C solution &
distilled
water separately . . . . . . . . . . . . . . . 3
pcs
11. Metallic test-tube rack
1 pc
12. 10-ml measuring cylinder
1 pc
13. 250-ml beaker for discarding waste
1 pc
14. 5-ml pipette 1
pc
15. Pipette filler . . . . . . . . . . . . . . . . 1
pc
16. Teat pipette 1
pc

On each students’ bench:

17. Lemon 1/3 pc
18. Green lantern pepper 1/3
pc

On teacher’s bench:
19. 20 ml DCPIP solution
20.  ml standard vitamin C solution (of known concentration)
21. Balance
22. Weighing paper 2
pcs

On side bench:
23. Distilled water
End of handout