EXPERIMENT NO.

5
1 Plant extracts contain molecules which have many uses in industry and as medicines.

It is important for scientists to be able to estimate the concentration of these molecules in plant
extracts.

You will estimate the concentration of molecule R in a sample of plant extract U.

You are provided with the materials shown in Table 1.1.

Table 1.1

labelled contents hazard volume / cm3

10R 10% solution of molecule R none 100

W distilled water none 100

U unknown concentration of none 40

molecule R in a plant extract
A sulfuric acid solution harmful 20
irritant
K potassium manganate(VII) solution irritant 20

If any of the solutions come into contact with your skin, wash off immediately under cold water.

It is recommended that you wear suitable eye protection and wear gloves to protect your hands
when using A and K.

9700/31/M/J/22
(a) You will need to carry out a serial dilution of the 10% molecule R solution, 10R, to reduce the
concentration by half between each successive dilution.

You will need to prepare four concentrations of molecule R solution in addition to the 10%
molecule R solution, 10R.

After the serial dilution is completed, you will need to have 10 cm3 of each concentration
available to use.

(i) Complete Fig. 1.1 to show how you will prepare your serial dilution.

Fig. 1.1 shows the first two beakers you will use to make your serial dilution. You will
need to draw three additional beakers.

For each beaker add labelled arrows to show:

• the volume of molecule R solution transferred

• the volume of distilled water, W, added.

Under each beaker, state the concentration of molecule R solution.

 0 cm3 of W

20 cm3 of
10.0% R, 10cms of
.............................
%
10

10R .............................

.............................

.............................
cocm
of

!
I&
of 5
locm" .

%
%

10 cm3 of
10.0% R
solution to
use 10cmqW ocm3
f2 R
.
5%

cit
10cm
of 5 0% R
.............................
.
10cm
first

I
Solution to use
--- ~

.............................
-

of
locm
=
=.
-

.............................

.............................
f
solation
a
locms Sit -
-- -

X -

= =- -

loamof .e
!Si
- - ~
-

use - =
-

20cm3
f
0 . 625% R

solution to use

Fig. 1.1

[3]
Molecule R changes an acidified potassium manganate(VII) solution, K, from a pink colour to
colourless.

The lower the concentration of molecule R, the greater the volume of R needed to change the
potassium manganate(VII) from pink to colourless (the end-point).

Carry out step 1 to step 12.

step 1 Prepare the concentrations of molecule R solution, as decided in (a)(i), in the beakers
provided.

step 2 Label 5 test-tubes with the concentrations of R you prepared in step 1.

step 3 Put 1 cm3 of A into the test-tube labelled 10%.

step 4 Put 1 cm3 of K into the same test-tube and mix well.

step 5 Put the nozzle of a clean 10 cm3 syringe into the beaker containing 10% R.

step 6 Pull the plunger out to the 5 cm3 mark so that 5 cm3 of 10% R enters the syringe.

step 7 Put the syringe containing 10% R into the test-tube from step 4 as shown in Fig. 1.2.

syringe resting on
top of test-tube

drop of solution
5 cm3 of solution
containing molecule R
containing molecule R

1 cm3 of A and 1 cm3 of K

Fig. 1.2

You will be counting the number of drops needed to change K from pink to colourless (the
end-point).

step 8 Gently press the plunger to add one drop of solution containing molecule R.

step 9 Mix well.

step 10 If the mixture stays pink repeat step 8 and step 9. Count the number of drops released to
reach the end-point. If the number of drops needed to reach the end-point is more than
100, record as ‘more than 100’.

step 11 Record the results in (a)(ii).

step 12 Repeat step 3 to step 11 for each of the concentrations of molecule R prepared in step 1.
 (ii) Record your results in an appropriate table.

Concentration
f R solution Number
of drops
(%)

L
(O

5 23

2 .
5 39

58
1 .

25

0 625 .
&2

[5]

(iii) State two significant sources of error when measuring the dependent variable in this
investigation.

Drop
In varies
...........................................................................................................................................
size .

The end point

q-D
fficult
to
judge
...........................................................................................................................................
.

...........................................................................................................................................

...........................................................................................................................................

..................................................................................................................................... [2]

(iv) Suggest how you could improve the procedure to reduce one of the errors stated in
(a)(iii).
the instead
1 . Measure voume used
of number
of drops
........................................................................................................................................... .

OR 2 - Use a colorimeter to
judge
the endpoint
...........................................................................................................................................
.

..................................................................................................................................... [1]

step 13 Repeat step 3 to step 10 with U, the unknown concentration of molecule R.

step 14 Record your result in (a)(v).

 (v) 48
State the number of drops of U needed to reach the end-point .................................... [1]

(vi) Complete Fig. 1.3 to show the positions of each of the percentage concentrations of
molecule R solution recorded in (a)(ii).
[1]
U

0.0% 10.0%
1 ↓
0 625% 1 25% 2 5% 5 0%
.
. .

percentage concentration of molecule R

Fig. 1.3

(vii) Use your results in (a)(ii) and (a)(v) to estimate the concentration of molecule R in
solution U.

Show this estimate of the concentration of U on Fig. 1.3, by placing the letter U on the
line in the correct position.
[1]
(b) A student investigated the concentration of molecule R in extracts from different plants.

The student added a fixed volume of the extract to acidified potassium manganate(VII) and
timed how long it took to reach the end-point.

The results are shown in Table 1.2.

Table 1.2

plant extract time / s

B 15
C 59
D 42
E 10
F 7

(i) Plot a bar chart of the data shown in Table 1.2 on the grid in Fig. 1.4.

Use a sharp pencil.

60
-

50
-

40-

30
-

f 20-

10
-

O -

C D E F
B
Plant Extract
Fig. 1.4
[4]
 (ii) Suggest which plant extract would contain the highest concentration of R.

Plant extract F
......................................................... [1]

(c) Scientists have suggested that a different molecule in the plant extract might act as an
antibiotic.

The scientists tested this by:

• spreading bacteria over the surface of agar gel containing nutrients

• putting small drops (3 μm3) of different concentrations of the plant extract into
separate wells in the agar gel
• incubating the agar gel for 20 hours
• measuring the inhibition area (where the bacteria were not observed) for each
concentration of plant extract.

The results are shown in Fig. 1.5.

120

100

9G

80

inhibition
60
area / mm2

40

20

0
0 20 40 60 80 100
22 concentration of solution
of molecule R / μg cm–3

Fig. 1.5
(i) Use the graph in Fig. 1.5 to estimate the concentration of plant extract that results in an
inhibition area of 92 mm2.

22
ngcm-
concentration of plant extract = ......................................................... [1]

(ii) Suggest how the molecule in the plant extract may inhibit the growth of the bacteria.

- Prevent formation of cell membrane

...........................................................................................................................................
1
.

Bacterial cell
8
Lysis
...........................................................................................................................................
.
-

Inhibits protein synthesis

...........................................................................................................................................
3 -
.

Inhibits cell division.

...........................................................................................................................................
4 -

..................................................................................................................................... [2]

Cary two]
[Total: 22]