NANYANG JUNIOR COLLEGE H2 Biology (Practical)

Term 3 Week 3

Name: Class: Date:

PRACTICAL 10 : Effect of starch concentration on diastase activity

1 Enzyme E catalyses the hydrolysis of starch to maltose.

The end-point of the reaction can be found by measuring the time taken for all the starch to
be hydrolysed.

You will investigate the effect of the independent variable, starch concentration, on enzyme
E.

You are provided with the materials shown in Table 1.1.

Table 1.1

labelled contents hazard volume / cm3

E 1% diastase solution irritant 10

S 0.5% starch solution none 30

W distilled water none 100

iodine iodine solution irritant 30

unknown concentration of starch 10

U none
solution

If any solution comes into contact with your skin, wash off immediately under cold water.

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 NANYANG JUNIOR COLLEGE H2 Biology (Practical)
Term 3 Week 3

(a) You are required to investigate this hypothesis by carrying out a serial dilution of starch
solution which reduces the concentration by half between each successive dilution.

Fig. 1.1 shows how to make the first concentration of 0.25% starch solution.
3 3
10 cm of distilled 3 10 cm of distilled
water, W 10 cm of distilled water, W
water, W 3
3
3
10 cm of 0.25% 10 cm of 0.125%
10 cm of starch starch solution starch solution
solution, S

0.125% 0.0625%
3
30 cm of 0.5% 0.25% starch
starch solution, S solution

Fig. 1.1

(i) Complete Fig. 1.1 to show how you will make two further concentrations of starch
solution. [3]
C - 1m for correct diluted concentration – 0.125 AND 0.0625 + units % at least once;
S - 1m for addition of starch solution to both beakers (e.g. 10 cm3 of 0.025 beaker
AND 10 cm3 of 0.125 to next beaker);
W- 1m for addition of distilled water to both beakers (e.g. 10 cm3 of water);

Carry out step 1 to step 15.

1. Prepare the concentrations of starch solution as shown in Fig. 1.1 in the beakers
provided.
2. Label test tubes with the concentrations of starch solutions.

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 NANYANG JUNIOR COLLEGE H2 Biology (Practical)
Term 3 Week 3

3. Wipe the spotting tile clean with a damp paper towel and then dry the tile. Label the tile,
as shown in Fig. 1.2. The numbers indicate the sampling times in seconds.

Fig. 1.2
3
4. Using a 2 cm syringe, put two drops of iodine on the tile at each sampling time, as
shown in Fig. 1.2.
5. Put 3.0 cm3 of S into the labelled test-tube.
6. Put 0.5 cm3 of E into the same test-tube. Use a glass rod to mix.
7. Start timing immediately.
8. After 15 seconds, use the glass rod to transfer a drop of the mixture from the test tube
onto the iodine that is labelled 15, on the spotting tile.
9. Immediately clean the glass rod with a paper towel.
10. Repeat steps 8 to 9 at 15 second intervals until the iodine does not change colour. This
is the end-point.
11. Record the time taken to reach the end-point in an appropriate table in (a)(ii). If the
iodine changes colour at 180 seconds, record ‘more than 180’.
12. Repeat steps 3 to 11 replacing the 3.0 cm3 of S with 3.0 cm3 of the other concentrations
of starch solution.

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 NANYANG JUNIOR COLLEGE H2 Biology (Practical)
Term 3 Week 3

(ii) Record your results in an appropriate table.

Concentration of starch /% Time taken to reach end-point /s

0.5 More than 180
0.25 135
0.125 60
0.0625 30

1. H - heading for independent variable: Concentration of starch / % and before heading for
dependent variable and no units in body of table ;
heading dependent variable: Time taken to reach end-point / s (reject sec or seconds)
and no units in body of table ;
Heading follows what’s stated in qns, reject other variations “time taken for iodine to
remain yellow brown / iodine to not change colour”.
2. R - readings for all samples (in 15s intervals) + result of 0.5% is more than 180 (reject
>180, follow qns) and result of 0.25% is 105-180s ;
3. T - Trend: time taken to reach end point increases with increasing starch concentration ;
4. S - results of seconds recorded to nearest whole second ;

Qns did not ask for mean (conduct of repeats) or rate of reaction.
[4]
13. Label a test‑tube with the letter U.
14. Repeat steps 3 to 11 with U instead of S.
15. Record in (a)(iii) your result for U.

(iii) State the result for U.

result for U ......................................................... [1]
records result for U and seconds ;
(iv) Using your results in (a)(ii) and (a)(iii), estimate the concentration of starch in U.
0.25% or 0.25 - 0.5% or 0.125 – 0.25% ; [1]
(v) State one significant source of error in this investigation.

Explain why this is a source of error.

source of error

explanation
any one from:
1 difficulty in judging exact time of colour change ;
2 colour may change between time intervals ;
3 using glass rod (apparatus needs to be mentioned) causes drop sizes of sample
(reject iodine as syringe is used) to be of unequal sizes / volumes (reject amount as
this is vague);
[1]

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 NANYANG JUNIOR COLLEGE H2 Biology (Practical)
Term 3 Week 3

Reject the following points:

1. cross contamination resulting in wrong starch concentration
2. time lag in transferring drop of solution onto tile at each sampling time
3. temperature for each starch concentration is different
4. mention of color standards e.g. insufficient color standards
5. lack of repeats
6. drop size taken up by glass rod too small / insignificant
7. glass rod not wiped properly
8. inconsistent stirring of mixture
9. drops of iodine added having inconsistent volumes (syringe is used to
standardise the drop size)
10. iodine oxidised before mixture is added

Sources of error do NOT refer to mistakes made when conducting the experiment.
Sources of errors are unavoidable limitations of the apparatus, measuring instruments,
experimental technique or experimental design that prevent the accuracy of the results.

(vi) Suggest how you could make one improvement to the independent variable so that a
more accurate estimate of the concentration of starch in U can be obtained.

any one from :

more concentrations between the two concentrations that lie each side of the
estimate + give 2 values for eg between 0.125 and 0.25% ; reject between 0.5 and
0.0625%
AVP ; @ the idea that more INTERMEDIATE concentrations is required. ;

® wider range (vague) / burette /wash apparatus with starch

[1]
(vii) Suggest how you could make two improvements to the measuring of the
dependent variable so that a more accurate estimate of the concentration of starch
in U can be obtained.

any two from:

1 use colorimeter ;
2 standard colours to compare ;
3 more intermediate times ;
4 choose end point that is dark brown ;
5 use pipette / syringe for sample; @ syringe ® dropper

® using syringe for iodine (already used) / equilibration / magnetic stirrer /

repeats  reliability, not accuracy / less drops / increase sampling time

[2]

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 NANYANG JUNIOR COLLEGE H2 Biology (Practical)
Term 3 Week 3

Part II
Proteins can be affected by presence of metal ions such as Cu2+ or Pb2+.

Table 1.1 shows the results of an investigation into the effect of the concentration of copper
sulfate solution on a protein suspension. A protein suspension was mixed with different
concentrations of copper sulfate.
After a set time, the percentage absorbance of light was measured using a colourimeter.

Table 1.1

Copper sulfate absorbance of light by protein suspension / %

concentration
/ mol dm-3 trial 1 trial 2 trial 3 trial 4 trial 5 mean
x 10-3
25.0 100 99 100 99 100 100
12.5 97 95 80 97 94 96
5.5 78 81 79 82 80 80
3.5 84 59 58 58 62
1.5 9 11 10 9 8 9

(a) (i) Draw a circle around each of the anomalous results and complete the table. [2]

12.5  80 AND 3.5  84 ;

Correct mean: 59 ;

Note to teachers: Anomalous data usually occur during ACE part, not the MMO part of
practical (i.e. students’ results based on experiment usually don’t contain anomalous
results)

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 NANYANG JUNIOR COLLEGE H2 Biology (Practical)
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(ii) Plot a graph of the data shown in Table 1.1.

Correct axis with units: x-axis (copper sulfate

concentration / moldm-3 x10-3) AND y-axis (mean
absorbance of light by protein suspension /%) ;

Appropriate scale:
x-axis  5.0 to 2 cm, label each 2cm
y-axis  20 to 2 cm, label each 2cm
label 0

correct plotting of each point to within half a

square;

best fit curve, no extrapolation ;

Reject point to point as mean value is used so there is a relationship between the
variables [4]

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 NANYANG JUNIOR COLLEGE H2 Biology (Practical)
Term 3 Week 3

(iii) Explain the effect of copper sulfate solution on protein suspension. Effect of heavy
metal on protein

Max 2:

1. (protein) coagulates / clots as concentration of copper sulfate increases ;

2. denatures;

3. detail on the effect of changing the protein structure ; eg bonds broken/ disrupted
(heavy metal cations form bond with negatively charged R groups of proteins and
disrupt ionic bonds) + loss/ distortion of unique 3D (spell out) conformation / altered
quaternary or tertiary structure ;

Reject any link to enzyme

[2]
[Total: 21]

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