New Senior Secondary Mastering Biology (Second Edition)

Practical Workbook for SBA 1A

Suggested answers to Practical Workbook

Ch 1 Introducing biology

Practical 1.1 Design an investigation of the effect of fresh

pineapple on the setting of jelly

Aim (p. 1-2)
To investigate the effect of fresh pineapple on the setting of jelly.

Introduction (p. 1-2)

1 Problem
Does the presence of fresh pineapple prevent the jelly from setting?

2 Hypothesis
The presence of fresh pineapple prevents the jelly from setting.

3 Principle
Prepare two identical jelly solutions. Add pieces of fresh pineapple to one of the jelly
solutions but not to the other. Leave the jelly solutions in the refrigerator for setting.

a Predictions
The jelly with fresh pineapple will not set. The jelly without fresh pineapple will
set.

b Identification of variables
i Presence of fresh pineapple. Cut a slice of fresh pineapple into small pieces and
put some pieces into one of the jelly solutions.

ii Whether the jelly sets or not. Tilt the container after leaving the jelly solution
in the refrigerator and observe if the solution has set or not.

iii Amount of jelly solution in each container, size and shape of the containers,
temperature of the jelly solutions, time of refrigeration, etc.

c Control
Yes. It is used to confirm that the presence of fresh pineapple is the only factor that

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prevents the jelly solution from setting.

d Assumptions
Other chemicals in the fresh pineapple do not affect the setting of jelly.

Materials and apparatus (p. 1-4)

Item Quantity
beaker (500 cm ) 3
1
measuring cylinder (100 cm3) 1
plastic container 2
glass rod 1
white tile 1
knife 1
fresh pineapple 1 slice
jelly powder
hot water
refrigerator 1
electronic balance 1

Procedure (p. 1-4)

1 Label two containers A and B.
2 Add 50 g of jelly powder and 200 cm3 of hot water to a beaker. Stir the mixture with a
glass rod until all the jelly powder dissolves.
3 Pour 100 cm3 of jelly solution into each container. Allow the jelly solutions to cool down
at room temperature.
4 Cut a slice of fresh pineapple into small pieces. Put some pieces into container A only.

5 Refrigerate the two containers overnight. Observe if the jelly solutions set.

Results (p. 1-5)

Container Content Does the jelly set?

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A Jelly solution + fresh pineapple No

B Jelly solution Yes

Discussion (p. 1-5)

1 The fresh pineapple may change the volume and the pH value of the jelly solution.

2 Use pineapple juice instead of pieces of fresh pineapple. Add 5 cm3 of fresh pineapple
juice to the jelly solution in the experimental set-up and add equal volume of distilled
water to the jelly solution in the control set-up.
Check the pH values of the jelly solutions in both set-ups. Add ethanoic acid to the jelly
solution in the control set-up drop by drop until the pH values of the jelly solutions in
both set-ups are the same.

3 Investigation of the minimum amount of fresh pineapple that can prevent the jelly
solution from setting. / Investigation of the effect of other fruits on the setting of jelly.

Conclusion (p. 1-6)

The presence of fresh pineapple prevents the jelly from setting.

Ch 2 The cell as the basic unit of life

Practical 2.1 Observation with a light microscope

Understanding procedure (p. 2-5)
1 To allow entry of a suitable amount of light. A dim image may result if there is
insufficient light while a faint image may result if the light is too bright.

2 The coarse adjustment knob leads to a larger degree of movement of the body tube. Any
downward movement of the body tube controlled by the coarse adjustment knob may
damage the objective or the slide because the distance between the objective and the
slide is very small.

Results (p. 2-5)

(Drawings vary with the cells observed. Drawings of human cheek cells and onion epidermal
cells are given as examples.)

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Questions (p. 2-6)

1
Low-power High-power
magnification magnification

Area of specimen observed

Larger Smaller
(smaller / larger)
Details of specimen
observed Less More
(more / less)
Brightness of image
Brighter Dimmer
(brighter / dimmer)

2 Move the slide to right. The image formed by the microscope is laterally inverted. The
actual movement of the Amoeba is towards the left.

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Practical 2.2 Preparation of temporary mounts of animal cells

and tissues
Understanding procedure (p. 2-8)
1 To stain the nuclei blue and act as a mounting medium.

2 To flatten the specimen so that they can be seen clearly in one plane of focus for the
objective lens. To prevent the objective lens from getting dirty by touching the specimen
or the mounting medium. To prevent the specimen from drying out by evaporation. To
protect the specimen from being damaged.

Results (p. 2-9)

Questions (p. 2-9)

1 Cell membrane, nucleus and cytoplasm.

2 Anywhere inside the cell.

3 No. / Yes, they have small vacuoles.

Practical 2.3 Preparation of temporary mounts of plant cells and

tissues
Understanding procedure (p. 2-14)
1 Hydrilla leaf cells have pigments which can be observed clearly under a microscope.
Onion epidermal cells and banana cells do not have pigments. They need to be stained
with iodine solution before observation.

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2 To spread out the cells so that light can pass through them.

Results (p. 2-15)

Questions (p. 2-16)

1 Cell wall, cell membrane, nucleus, cytoplasm, chloroplast and granule.

2 Near the side of the cell.

3 No. Not all of them contain chloroplasts or chlorophyll. Only those with chloroplasts are
green.

4 Similarities: Both of them have a nucleus / a cell membrane / cytoplasm. (any 2)

Differences: Plant cells are often larger than animal cells. / Plant cells have a definite
shape while animal cells do not. / Plant cells have a cell wall and some have chloroplasts.
Animal cells do not have cell walls or chloroplasts. / Plant cells usually have a large
vacuole while animal cells do not. (any 2)

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Ch 3 Movement of substances across cell membrane

Practical 3.1 Demonstration of osmosis using dialysis tubing

Understanding procedure (p. 3-3)
1 To give a more obvious change in the liquid level in the capillary tube.

2 Sucrose solution on the outside of the tubing will affect the result.

3 To show that any change in the liquid level in the experimental set-up is due to the
sucrose solution inside the dialysis tubing.

Results (p. 3-3)

Set-up Change in the liquid level in the capillary tube

Experimental Rises

Control Falls

Discussion (p. 3-4)

a There is a net movement of water molecules from distilled water to sucrose solution due
to osmosis.

b Water molecules move out of the tubing. This is because the hydrostatic pressure of the
liquid column forces water to move out of the tubing.

Questions (p. 3-4)

a The liquid level will rise faster and higher.

b The liquid level will fall and the tubing will finally shrink.

Practical 3.2 Demonstration of osmosis using living animal tissue

Understanding procedure (p. 3-6)
1 Set-up B is a control. It shows that any change in the liquid level in set-up A is due to the
concentrated sucrose solution inside the thistle funnel.

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2 To give a more obvious change in the liquid level in the tube of the thistle funnel.

Results (p. 3-7)

Set-up Change in the liquid level in the thistle funnel

A Rises

B Falls

Discussion (p. 3-7)

Distilled water has a higher water potential than concentrated sucrose solution, so there is a
net movement of water from the distilled water to the concentrated sucrose solution through
the differentially permeable animal tissue by osmosis. The volume of liquid inside the thistle
funnel increases and the liquid level rises.

Practical 3.3 Study of osmosis in red blood cells

Results (p. 3-9)
Concentration of
Appearance of the red blood cells
sodium chloride solution

0% Many red blood cells swell and burst.

0.45% Few red blood cells swell and burst.

0.9% Red blood cells are normal.

1.35% Few red blood cells shrink and become wrinkled.

1.8% Many red blood cells shrink and become wrinkled.

Questions (p. 3-9)

1 0.9% sodium chloride solution is isotonic to the red blood cells. At this concentration,
the red blood cells appear normal. It shows that there is almost no net movement of
water into or out of the cells because there is no difference in water potential between the
cells and their surroundings.

2 0% and 0.45% sodium chloride solutions are hypotonic to the red blood cells. At these
concentrations, the red blood cells swell and finally burst. It shows that the water

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potential of the red blood cells is lower than that of the sodium chloride solution. Water
enters the red blood cells by osmosis.

3 1.35% and 1.8% sodium chloride solutions are hypertonic to the red blood cells. At these
concentrations, the red blood cells shrink and become wrinkled. It shows that the water
potential of the red blood cells is higher than that of the sodium chloride solution. Water
leaves the red blood cells by osmosis.

Practical 3.4 Study of osmosis in living plant cells

Results (p. 3-12)
In concentrated In less concentrated In very dilute
sucrose solution sucrose solution sucrose solution

Discussion (p. 3-12)

1 The cytoplasm swells up gradually until the cell membrane presses tightly against the
cell wall.

2 No. This is because the water potential of each cell varies.

Practical 3.5 Study of osmosis in living plant tissue

Understanding procedure (p. 3-15)
1 Osmosis cannot take place across the potato peel because the peel is impermeable to
water. Any peel left on the potato discs will affect the results.

2 To prevent the evaporation of water which will change the concentration of the liquids in
the set-ups and affect the results.

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3 To remove the surplus water on the surface of the potato discs which will increase the
weight of the potato discs and affect the results.

4 To minimize the water loss from the potato discs by evaporation. Any water loss will
decrease the weight of the potato discs and affect the results.

Results (p. 3-15)

Title: Percentage change in average weight of potato discs in sucrose solutions of different
concentrations

Initial weight of Final weight of Percentage

Liquid
potato discs (g) potato discs (g) change in
inside
average
the tube Tube Average Tube Average weight (%)

A1 A1
Distilled
A2 A2
water
A3 A3

B1 B1
10% (Results
sucrose B2 vary with B2 (Results vary with Ss.)
solution Ss.)
B3 B3

C1 C1
20%
sucrose C2 C2
solution
C3 C3

Discussion (p. 3-16)

1 The potato discs in distilled water become heavier. This is because the water potential of
distilled water is higher than that of the potato cells. Water enters the potato discs by
osmosis.

2 The average weight of the potato discs in 10% sucrose solution changes very slightly.
This is because the water potential of 10% sucrose solution is nearly the same as that of
the potato cells. There is almost no net movement of water into or out of the potato discs.

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3 The potato discs in 20% sucrose solution become lighter. This is because the water
potential of 20% sucrose solution is lower than that of the potato cells. Water leaves the
potato discs by osmosis.

Ch 4 Enzymes and metabolism

Practical 4.1 Demonstration of the breaking-down action of

enzymes
Understanding procedure (p. 4-3)
It is a control to show that no oxygen is given off from hydrogen peroxide solution without
tissue.

Results (p. 4-3)

Tube Tissue Glowing splint relights

A potato +

B apple +

C liver +

D meat +

E no tissue added –

Discussion (p. 4-3)

1 Tissues of potatoes, apples, liver and meat catalyse the breakdown of hydrogen peroxide
to oxygen.

2 Boiled tissues, instead of fresh tissues, can be used in a further investigation. If the
boiled tissues have no catalytic action, it is more likely that the reaction is catalysed by
an enzyme.

Practical 4.2 Demonstration of the building-up action of enzymes

Understanding procedure (p. 4-7)
1 To break the potato cells and release the enzyme from them.

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2 To prevent the enzyme in the potato tissue from denaturing by the heat produced during
grinding.

3 To remove starch present in the ground potato tissue. The starch can turn the iodine
solution blue-black and affect the results.

Results (p. 4-7)

Iodine solution
Tube
Original colour Final colour

A Brown Blue-black

B Brown Brown

C Brown Brown

Discussion (p. 4-7)

The mixture in tube A turns iodine solution blue-black. This is because starch phosphorylase
in potato tissue catalyses the synthesis of starch from glucose-1-phosphate. Therefore, starch
is formed in tube A.
No colour change is observed in tubes B and C (control set-ups) because no starch is formed
in these tubes.

Practical 4.3 Investigation of the effect of temperature on

enzyme activity
Understanding procedure (p. 4-10)
1 To ensure that the amylase and starch solutions inside the tubes reach the respective
temperatures before the reaction starts.

2 To prevent the changing of the condition of the mixture by any residue in the dropper.

Results (p. 4-10)

Time taken for the disappearance of
Temperature (°C)
blue-black colour (min)

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0 The blue-black colour does not disappear.

20

40 (Results vary with the origin of amylase.)

60

80

100 The blue-black colour does not disappear.

Discussion (p. 4-11)

Amylase is inactive at low temperatures. Its activity increases with temperature and is the
highest at 60 °C. Afterwards the activity decreases and stops at 100 °C. With a rise in
temperature, the kinetic energy of amylase and starch molecules increases. The molecules
collide and react more frequently. As the temperature increases further, the enzyme is
denatured and the reaction rate decreases. At 100 °C, all amylase is denatured and no reaction
takes place.

Extended questions (p. 4-11)

a Starch would be broken down and blue-black colour would disappear. This is because
the inactive amylase would resume its activity with a rise in temperature.

b Starch would not be broken down and blue-black colour would remain. This is because
the activity of the denatured amylase would not restore even when it is cooled.

Conclusion (p. 4-11)

Amylase is inactive at low temperatures. Its activity increases with temperature until it
reaches a maximum. Afterwards the activity decreases and stops.

Practical 4.4 Design an investigation of the effect of pH on

enzyme activity
Aim (p. 4-13)
To study the effect of pH on the activity of amylase.

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Introduction (p. 4-13)

1 Problem
What is the effect of pH on enzyme activity?

2 Principle
To study the effect of pH on enzyme activity, starch-agar plates can be used. When
incubated with filter paper discs soaked with buffer solution at different pH values and
amylase, the starch-agar in the plate will be broken down and clear zones will be
observed. The larger the diameter of the clear zone, the higher the amylase activity.

a Identification of variables
i pH of the solutions. Add buffer solutions at different pH into the solutions.

ii Diameter of the clear zone on the starch-agar plate. Measure the diameter of
the clear zone with a piece of graph paper.

iii Volumes of buffer and amylase solutions added into the wells of the spot plate,
time and temperature for incubating the starch-agar plate, etc.

b Control
No. The investigation aims to study the enzyme activity at different pH values.

c Assumptions
The diameter of the clear zone on the starch-agar plate has a positive correlation
with the enzyme activity.

Materials and apparatus (p. 4-14)

Item Quantity
starch-agar plate 1
forceps 1 pair
filter paper 1
dropper 7
spot plate 1
graph paper 1
marker pen 1
paper hole punch 1
disposable gloves 1 pair
incubator 1

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citrate-phosphate buffer solution at different

pH values (pH 5, 6, 7, 8 and 9)
0.1% amylase solution
iodine solution
distilled water

Procedure (p. 4-15)

1 Label five wells of a spot plate A to E. Add the following solutions into the wells.
Well Solution
1 drop of amylase solution + 1 drop of citrate-phosphate buffer solution at pH
A
5
1 drop of amylase solution + 1 drop of citrate-phosphate buffer solution at pH
B
6
1 drop of amylase solution + 1 drop of citrate-phosphate buffer solution at pH
C
7
1 drop of amylase solution + 1 drop of citrate-phosphate buffer solution at pH
D
8
1 drop of amylase solution + 1 drop of citrate-phosphate buffer solution at pH
E
9
2 Prepare five filter paper discs using a paper hole punch. Put a paper disc into each well.
3 Label the side of a starch-agar plate A to E. Remove the discs from the wells using a pair
of forceps. Drain away any excess solution from the discs. Place the paper discs onto the
plate according to the labels.
4 Cover the plate with the lid. Incubate the plate at 35 °C for one hour.
5 Remove the paper discs from the plate and flood the plate with iodine solution. Leave it
for one minute.
6 Rinse the plate with distilled water.
7 Place the plate on a piece of graph paper and examine against light. Measure the
diameter of the clear zones around the areas covered by the paper discs.

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Results (p. 4-15)

Diameter of the clear zone
Well Solution (number of squares on
graph paper)

1 drop of amylase solution + 1 drop of

A
citrate-phosphate buffer solution at pH 5

1 drop of amylase solution + 1 drop of

B
citrate-phosphate buffer solution at pH 6

1 drop of amylase solution + 1 drop of

C (Results vary with Ss.)
citrate-phosphate buffer solution at pH 7

1 drop of amylase solution + 1 drop of

D
citrate-phosphate buffer solution at pH 8

1 drop of amylase solution + 1 drop of

E
citrate-phosphate buffer solution at pH 9

Discussion (p. 4-16)

1 Amylase in the paper discs diffuses to the starch-agar and catalyses the breakdown of
starch. When iodine solution is added, the part of the starch-agar plate containing starch
turns blue-black while the part without starch becomes clear. Therefore, clear zones are
formed in the starch-agar.

2 (Answer depends on the results.) (Optimum pH of amylase: 5.6 – 6.9)

Extended questions (p. 4-16)

a No clear zone would be formed. This is because extremely low pH would denature the
amylase and stop its activity.

b No clear zone would be formed. This is because extremely high pH would also denature
the amylase and stop its activity.

Conclusion (p. 4-16)

The activity of amylase is the highest at pH 6 (depends on the results). It slows down or even
stops at extreme pH values.

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Practical 4.5 Investigation of the effect of inhibitors on enzyme

activity
Understanding procedure (p. 4-18)
It is a control to show that the activity of urease is slowed down or stopped by mercuric ions.

Results (p. 4-19)

Colour of bromothymol blue indicator
Tube
Original colour Final colour

A Yellow Yellow

B Yellow Blue

Discussion (p. 4-19)

No colour change is observed in tube A. Mercuric ions are an inhibitor of urease. The activity
of urease slows down or even stops in the presence of mercuric ions. Therefore, no ammonia
is formed in tube A. In tube B, urease catalyses the breakdown of urea into ammonia in the
absence of mercuric ions. The ammonia turns the bromothymol blue indicator blue.

Conclusion (p. 4-19)

Mercuric ions slow down or even stop the activity of urease.

Practical 4.6 Investigation of the effectiveness of different

biological washing powders

Results (p. 4-22)
Tube Change in the size of the egg white cube

B (Results vary with Ss.)

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Discussion (p. 4-22)

1 (Answer depends on the results.)

2 No. We should also consider the price of each brand.

3 Use more washing powder to prepare the washing powder solution.

Extended questions (p. 4-22)

It is because proteins in silk and wool will be broken down by the proteases.

Conclusion (p. 4-23)

(Answer varies with the results.)

Practical 4.7 Design an investigation of protease activities in

different fruit juices

Aim (p. 4-25)
To compare the protease activities in different fruit juices.

Introduction (p. 4-25)

1 Problem
Which fruit juice has the highest protease activity?

2 Principle
To compare the protease activities in different fruit juices, milk-agar plates can be used.
When incubated with fruit juices containing protease, the milk protein in the plate will be
broken down and clear zones will be formed. The larger the diameter of the clear zone,
the higher the protease activity.

a Identification of variables
i Kinds of fruit juices. Add different fruit juices into different wells of the milk-
agar plate.

ii Diameter of the clear zone formed on the milk-agar plate. Measure the
diameter of the clear zone around the well with a piece of graph paper.

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iii Volumes of juice added into the wells of the plate, time and temperature for
incubating the milk-agar plate, etc.

b Control
Yes. It is used to confirm that the formation of clear zones is due to the fruit juices.

c Assumptions
Other chemicals in the fruit juices would not affect the formation of clear zones. /
The diameter of the clear zone has a positive correlation with the enzyme activity.

Materials and apparatus (p. 4-27)

Item Quantity
cork borer 1
Bunsen burner 1
insulating mat 1
dropper 5
test tube 5
test tube holder 1
milk-agar plate 1
graph paper 1
marker pen 1
incubator 1
pineapple juice
kiwi fruit juice
papaya juice
guava juice
distilled water

Procedure (p. 4-27)

1 Heat the end of a cork borer in a Bunsen flame and allow it to cool.
2 Gently press the borer down into the milk-agar to make five wells. Replace the lid as
quickly as possible.
3 Label the wells A to E by marking on the side of the Petri dish.
4 Use five clean droppers to fill up wells A to E with pineapple juice, kiwi fruit juice,
papaya juice, guava juice and distilled water respectively.

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5 Replace the lid. Incubate the plate at 35 °C for one hour.

6 Place the plate on a piece of graph paper and examine against light. Measure the
diameter of the clear zones around the wells.

Results (p. 4-28)

Diameter of clear zone
Well Sample
(number of squares on graph paper)

A Pineapple juice

B Kiwi fruit juice

C Papaya juice (Results vary with Ss.)

D Guava juice

E Distilled water

Discussion (p. 4-28)

1 Protease in the fruit juices diffuses to the milk-agar and catalyses the breakdown of the
white milk protein. Therefore, clear zones are formed around the wells containing fruit
juices with protease.

2 (Answer depends on the results.)

3 (Answer depends on the results.)

Conclusion (p. 4-28)

(Answer depends on the results.)

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Ch 5 Food and humans

Practical 5.1 Detection of food substances by food tests

Understanding procedure (p. 5-5)
1 No. This is because the food tests are qualitative tests to show the presence of certain
food substances. They are not quantitative tests.

2 The water bath provides a better control over the temperature and can prevent bumping
of the mixture.

Results (p. 5-5)

A Test for glucose using glucose test paper
Colour of the test end of the test paper
Sample
Original colour Final colour

(Results vary with the type

Glucose solution
of the test paper used.)

Distilled water

B Test for reducing sugars using Benedict’s test

Change of the mixture of
Sample
Benedict’s solution and sample

Glucose solution Brick-red precipitate is formed

Distilled water No observable change

C Test for starch using iodine test

Sample Colour of iodine solution

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Original colour Final colour

Starch solution Brown Blue-black

Distilled water Brown Brown

D Test for lipids using grease spot test

Is a translucent spot present after drying?
Sample Before immersing into After immersing into
organic solvent organic solvent

Cooking oil Yes No

Distilled water No No

E Test for proteins using protein test paper

Colour of the test end of the test paper
Sample
Original colour Final colour

(Results vary with the type

Egg white solution
of the test paper used.)

Distilled water

F Test for vitamin C using DCPIP solution

Sample Colour of DCPIP solution

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Original colour Final colour

Vitamin C
Blue Colourless
solution

Distilled water Blue Blue

Discussion (p. 5-6)

1 The translucent spot caused by lipids is permanent. On the contrary, the translucent spot
caused by water disappears when water evaporates.

2 Lipids dissolve in the organic solvent. Therefore, the translucent spot caused by lipids
disappear.

Question (p. 5-7)

As the red colour of the blood will mask the results of the Benedict’s test, the blood sample
should be diluted with distilled water first. Alternatively, the blood sample should be
centrifuged and the plasma collected is used to perform the Benedict’s test.

Practical 5.2 Investigation of the food substances in common

foodstuffs
Results (p. 5-11)
Food Reducing
Glucose Starch Lipids Proteins Vitamin C
sample sugars

(Results vary with the types of foods tested.)

Discussion (p. 5-11)

(Answer depends on the types of foods tested.)

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Practical 5.3 Design an investigation to compare the amount of

vitamin C in different fruits and vegetables

Aim (p. 5-13)
To compare the vitamin C content in different fruits and vegetables.

Introduction (p. 5-13)

1 Problem
Which type of fruit or vegetable has the highest vitamin C content?

2 Principle
Vitamin C is a reducing agent. When it is added to DCPIP solution, the DCPIP is reduced
and decolourized. The relative vitamin C content in juices of different kinds of fruits and
vegetables can be found by comparing the amounts of juices required to decolourize a
fixed amount of DCPIP solution.

a Identification of variables
i Kinds of fruits and vegetables. Add juices of different kinds of fruits and
vegetables into the DCPIP solution.

ii Vitamin C content in juices. Measure the number of drops of juice sample

needed to decolourize a fixed amount of DCPIP solution.

iii Volume and concentration of DCPIP solution used with each sample,
temperature of samples and DCPIP solution, etc.

b Control
Yes. It is used to confirm that the decolourization of DCPIP solution is due to the
juices.

c Assumptions
The decolourization of DCPIP solution is due to the vitamin C only. / No other
reducing agents are present in the juice to decolourize the DCPIP solution.

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Materials and apparatus (p. 5-15)

Item Quantity
test tube 12
test tube rack 1
dropper 7
measuring cylinder (10 cm3) 1
mortar and pestle 1
knife 1
filter funnel 1
fine muslin or filter paper
0.02% DCPIP solution
distilled water
orange, lemon, green pepper,
broccoli, guava, kiwi fruit

Procedure (p. 5-15)

1 Cut the fruit or vegetable into small pieces.
2 Put the small pieces into a mortar and grind them with a small known volume of cool
distilled water (if necessary) using the pestle.
3 Squeeze the ground materials through several layers of pre-moistened fine muslin or
filter them through a filter paper. Collect the filtrate (i.e. the juice). Skip this step if a fine
and fairly colourless suspension is obtained.
4 Put 1 cm3 of DCPIP solution into a test tube.
5 Use a dropper to add the juice, drop by drop, to the DCPIP solution until the solution is
decolourized. Record the number of drops of juice added.

6 Repeat steps 4 and 5 with the juices extracted from different fruits and vegetables. If the
decolourization is too quick (i.e. the juice is too concentrated), dilute the juice by a
known volume of distilled water and repeat steps 4 and 5. Take this dilution factor into
consideration in the comparison of vitamin C content.
7 Repeat steps 4 and 5 with distilled water instead of juice as a control.

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Results (p. 5-16)

Number of drops of sample
Relative vitamin C
Sample needed to decolourize 1 cm3
content
of DCPIP solution

Orange

Lemon

Green pepper (Results vary with Ss.)

Broccoli

Guava

Kiwi fruit

Distilled water

Discussion (p. 5-16)

1 (Answer depends on the results.)

2 (Answer depends on the results.)

3 Reducing property

4 The vitamin C in the juices is oxidized by air. / The observation of complete

decolourization of DCPIP solution is subjective, especially when the juices are
coloured. / There are other reducing agents in the juices.

5 Prepare the juices quickly to prevent the oxidation of vitamin C.

Use fruits or vegetables with light-coloured juices.

6 Fruits and vegetables with dark-coloured juices cannot be used. The dark colour of the
juices may mask the colour of the DCPIP solution.

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Practical 5.4 Design an investigation to study the effect of boiling

on the amount of vitamin C in vegetables

Aim (p. 5-19)
To study the effect of boiling on the vitamin C content in vegetables.

Introduction (p. 5-19)

1 Problem
Does boiling reduce the vitamin C content in vegetables?

2 Principle
Vitamin C is a reducing agent. When it is added to DCPIP solution, the DCPIP is
reduced and decolourized. The relative vitamin C content in juices of boiled vegetable
and unboiled vegetable can be found by comparing the amounts of juices required to
decolourize a fixed amount of DCPIP solution.

a Identification of variables
i Whether the vegetable is boiled. Boil some vegetable samples in water and
leave some samples unboiled.

ii Vitamin C content in the juice. Measure the number of drops of the juice
needed to decolourize a fixed amount of DCPIP solution.

iii Kind of vegetable, temperature of the juice and DCPIP solution, etc.

b Control
Yes. It is used to confirm that boiling is the only factor that reduces the vitamin C
content in vegetables.

c Assumptions
The decolourization of DCPIP solution is due to the vitamin C only. / No other
reducing agents are present in the juice to decolourize the DCPIP solution.

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Materials and apparatus (p. 5-20)

Item Quantity
test tube holder 1
test tube 6
test tube rack 1
dropper 3
measuring cylinder (10 cm ) 3
2
mortar and pestle 1
water bath 1
knife 1
safety goggles 1 pair
filter funnel 1
filter paper
0.02% DCPIP solution
distilled water
green pepper

Procedure (p. 5-21)

1 Cut a green pepper in half. Then, remove the seeds inside. Cut the one of the pieces into
small pieces.
2 Put five pieces of green pepper into a test tube containing 10 cm3 of distilled water. Then,
put the test tube into a boiling water bath for about 10 minutes. Remove the pieces of
boiled green pepper from the test tube.
3 Put the pieces of green pepper into a mortar and grind them with a small known volume
of cool distilled water using a pestle.
4 Squeeze the ground materials through a filter paper. Collect the filtrate (i.e. the juice).
5 Put 1 cm3 of DCPIP solution into a test tube.
6 Use a dropper to add the boiled juice, drop by drop, to the DCPIP solution until the
solution is decolourized. Record the number of drops of juice added.

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7 Repeat steps 3 to 6 with the other half of the unboiled green pepper. If the
decolourization is too quick (i.e. the juice is too concentrated), dilute the juice by a
known volume of distilled water and repeat steps 5 and 6. Take this dilution factor into
consideration in the comparison of vitamin C content.

Results (p. 5-21)

Number of drops of sample
Relative vitamin C
Sample needed to decolourize 1 cm3
content
of DCPIP solution

Boiled green
(Results vary with Ss.)
pepper

Unboiled green
pepper

Discussion (p. 5-22)

1 The vitamin C content in boiled green pepper is lower than that of unboiled green pepper
because vitamin C may be destroyed by high temperatures or may dissolve in the boiling
water.

2 Prepare unboiled vegetables (salad) instead of boiled vegetables.

3 The vitamin C in the juices is oxidized by air. / The observation of complete

decolourization of DCPIP solution is subjective, especially when the juices are
coloured. / There are other reducing agents in the juices.

4 Prepare the juices quickly to prevent the oxidation of vitamin C. / Use vegetables with
light-coloured juices.

5 Vegetables with dark-coloured juices cannot be used. The dark colour of the juices may

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 New Senior Secondary Mastering Biology (Second Edition)
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mask the colour of the DCPIP solution.

Conclusion (p. 5-22)

Boiling reduces the vitamin C content in vegetables.

Ch 6 Nutrition in humans

Practical 6.1 Examination of the mammalian digestive system

Questions (p. 6-2)

1
A Mouth cavity B Oesophagus

C Stomach D Duodenum

E Pancreas F Appendix

G Ileum H Caecum

I Liver J Colon

K Rectum L Anus

2 A  B  C  D  G 
J  K  L

3
Structure Nutrition process involved

A Ingestion, digestion

C Digestion

D Digestion, absorption

G Digestion, absorption

L Egestion

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Practical 6.2 Investigation of the action of pepsin

Understanding procedure (p. 6-6)
1 To provide an acidic medium for the action of pepsin.

2 To sterilize the end of the cork borer.

3 To minimize the exposure of the milk-agar plate to air and reduce the chance of
contamination.

4 Well B is a control to show that the formation of clear zones is due to pepsin. Well C is a
control to show that the formation of clear zones is not due to dilute hydrochloric acid.

Results (p. 6-7)

Presence of clear zone
Well Solution
around the well

pepsin solution +
A +
dilute hydrochloric acid

boiled pepsin solution +

B –
dilute hydrochloric acid

distilled water +
C –
dilute hydrochloric acid

Discussion (p. 6-7)

1 A clear zone is formed around well A. Pepsin in well A diffuses to the milk-agar and
helps digest the milk protein. Boiling denatures pepsin, so no clear zone is formed
around well B. Well C contains no pepsin, so no clear zone is formed around the well.

2 Peptides

Conclusion (p. 6-7)

Pepsin helps digest milk protein in an acidic medium.

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Practical 6.3 Demonstration of the effect of bile salts on oil

Results (p. 6-9)
Mixture Appearance of the mixture

Oil and
An emulsion is formed.
bile salt solution
Two layers of liquids can be seen: oil on the top
Oil and
and water at the bottom. The two liquids do not
distilled water
mix.

Discussion (p. 6-9)

Water cannot break down oil into tiny droplets as the bile salt solution does. Therefore, no
emulsion is formed and two layers of liquids can be seen.

Practical 6.4 Simulation of digestion and absorption in the small

intestine using dialysis tubing

Understanding procedure (p. 6-12)
1 Starch solution on the outside of the tubing will affect the result.

2 Less water allows a higher concentration of starch or reducing sugar molecules for easy
detection.

Results (p. 6-12)

Starch Reducing sugars

At start – –

After one hour – +

Questions (p. 6-13)

1
Part of the set-up Part of the human body

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Dialysis tubing Wall of the small intestine

Water surrounding the

Blood
dialysis tubing

Starch and amylase mixture Mixture of undigested food and digestive

in the dialysis tubing enzymes

2 a Reducing sugar (maltose)

b Amylase digests starch into maltose. Maltose molecules are small enough to pass
through the tubing and diffuse into the water outside the tubing, whereas the large
molecules (starch) are retained inside the tubing.

3 Through digestion, food substances are broken down into small molecules that can
diffuse through the wall of small intestine into the blood for use in our body.

4 Maltose molecules are not small enough to pass through the small intestine. / The small
intestine can absorb digested food by active transport but the dialysis tubing cannot. /
The small intestine can secrete enzymes but the set-up cannot. / The small intestine
shows peristalsis but the set-up does not. / There are many types of food molecules in the
small intestine apart from starch. / The food molecules have to pass through more than
one layer of cells instead of only one layer of tubing. / The blood is enclosed in blood
vessels. (any 3)

5 Diffusion rate of the reducing sugar molecules can be increased by stirring the
surrounding water and using a water bath at a higher temperature.
More concentrated solutions of starch and amylase can be used to speed up the reaction
rate.

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