EXPERIMENT NO.
2
1 Grapes are fruit that contain high concentrations of soluble sugars such as sucrose, fructose and
glucose.
The proportions of these sugars change as the grapes mature.
You will determine the concentration of reducing sugars in a sample of grape extract by using
known concentrations of reducing sugar.
You are provided with the materials shown in Table 1.1.
Table 1.1
labelled contents hazard volume / cm3
G grape extract none 20
W distilled water none 100
B Benedict’s solution harmful irritant 40
8.0% reducing sugar
R none 40
solution
If any solution comes into contact with your skin, wash off immediately under cold water.
It is recommended that you wear suitable eye protection.
You will need to carry out a serial dilution of the 8.0% reducing sugar solution, R, to reduce the
concentration by half between each successive dilution.
You will need to prepare four concentrations of reducing sugar in addition to 8.0% reducing sugar
solution, R.
After the serial dilution is completed, you will need to have 10 cm3 of each concentration available
to use.
(a) (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 reducing sugar solution transferred
• the volume of distilled water, W, added.
Under each beaker, state the concentration of reducing sugar solution.
9700/32/M/J/23
0 cm3 of W
20 cm3 of 8.0%
reducing sugar
solution, R
·I
10 cm3 of 8.0%
I
reducing sugar
solution to use 10cms
of 4 . 0 %
reducing
goo
10cms
of solution
reducing sugar
........................
%
of 1
reducingsagrse
0
........................ 10cm3
:
........................ 10cms
of W
-
=
%
of
10cms 2 :
0
.......
--
- - - -
- - -
-
-
. . .
- -
...
L Ocas
of
0 .
5%
sugar
reducing
solution to use
Fig. 1.1
[3]
Carry out step 1 to step 16.
step 1 Set up a water-bath and heat to boiling ready for step 6 and step 14.
step 2 Prepare the concentrations of reducing sugar solution as shown in Fig 1.1.
step 3 Label test-tubes with the concentrations shown in Fig. 1.1.
step 4 Put 2 cm3 of the 8.0% reducing sugar solution into the appropriately labelled
test-tube.
step 5 Put 2 cm3 of Benedict’s solution, B, into the same test-tube. Shake gently to mix.
step 6 Put this test-tube in the boiling water-bath. Start timing.
step 7 Measure the time taken to the first appearance of a colour change in the test-tube.
If there is no colour change after 120 seconds, stop timing and record as ‘more than 120’.
step 8 Record the result from step 7 in 1(a)(ii).
step 9 Remove the test-tube from the water-bath. Put the test-tube in the test-tube rack.
step 10 Repeat step 4 to step 9 with the remaining concentrations of reducing sugar solution.
(ii) Record your results in an appropriate table for the known concentrations of reducing
sugar.
Time taken to
first of
Concentration o appearance a
Sugar ( )
reducing
Colour
change
. (s)
& O .
4. O 12
2 O . 16
.O
1
21
0 5 .
28
[5]
(iii) Identify one source of error in step 7.
Difficult to
judge the
first colour
change .
...........................................................................................................................................
...........................................................................................................................................
..................................................................................................................................... [1]
(iv) To determine the concentration of reducing sugar in grape extract G, you will need to test
a sample of the extract.
State the volume of grape extract G that you will use to test for reducing sugars.
2cm3
volume = .......................................
[1]
step 11 Label a test-tube G.
step 12 Transfer the volume of grape extract G that you stated in (a)(iv) into test-tube G.
step 13 Put 2 cm3 of Benedict’s solution, B, into the same test-tube. Shake gently to mix.
step 14 Put this test-tube in the boiling water-bath. Start timing.
step 15 Measure the time taken to the first appearance of a colour change in the test-tube.
If there is no colour change after 120 seconds, stop timing and record as ‘more than 120’.
step 16 Record the result from step 15 in (a)(v).
(v) Record the time taken for the first colour change in test-tube G.
18S
time taken = ......................................................... [1]
(vi) The concentration of reducing sugars in G can be estimated from a graph of your results.
Draw a graph of the results you recorded in (a)(ii) on the grid in Fig. 1.2, using a line of
best fit.
The axes have been labelled for you.
Use a sharp pencil.
32-
(
20 - x
24-
time to first X
colour change 20
/s
18 -
16- X
12- *
& 16 i 6b
percentage concentration of reducing sugars
Fig. 1.2
[2]
(vii) Use your graph to estimate the percentage concentration of reducing sugars in G.
Show on your graph how you determined your answer.
6 % 1
percentage concentration of reducing sugars in G ................................................................
.
[2]
(viii) Suggest how you would modify this investigation to obtain a more accurate estimate for
the concentration of reducing sugars in sample G.
Repeat the test and
find the mean time .
...........................................................................................................................................
Use white card behind the test tube to
...........................................................................................................................................
a
the
see colour
change .
...........................................................................................................................................
..................................................................................................................................... [2]
(b) The concentration of reducing sugars in grapes changes as the grapes age (get older).
Table 1.2 shows the concentration of reducing sugars for grapes of different ages.
Table 1.2
age of grapes / days percentage concentration of reducing sugars
14 1.1
28 1.9
42 2.6
56 3.9
70 7.5
84 11.3
(i) Plot a graph of the data shown in Table 1.2 on the grid in Fig. 1.3.
12
j
10
e
8
·
Y
2
1 .
6
*
0
0 20 40 60 80 100
is of grapes/day
o
Fig. 1.3
[2]
(ii) Use your estimate from (a)(vii) and your graph in (b)(i) to estimate the age of the grapes
that were used to make grape extract G.
27
age of grapes ................................................ days [1]
(c) Grapes contain starch as well as reducing sugars. In a study, the concentration of amylase in
grapes was measured as the grapes aged.
The results of the study are shown in Fig. 1.4.
0.35
0.30
0.25
concentration of 0.20
amylase
/ arbitrary units 0.15
0.10
0.05
0.00
14 28 42 56 70 84
age of grapes / days
Fig. 1.4
Use the data in Fig. 1.3 and Fig. 1.4 to suggest a possible explanation for the change in the
concentration of reducing sugars in grapes as they age.
The concentration and
of amylase reducing sugar
...................................................................................................................................................
increases
...................................................................................................................................................
grapes
as
age
.
As
amylase hydrolyses starch into
reducing
...................................................................................................................................................
sugars
.
amylase means more
reducing
...................................................................................................................................................
More
sugar
.
...................................................................................................................................................
............................................................................................................................................. [3]
[Total 23]