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Biological Molecule Testing Syllabus

The document outlines an experiment to identify biological molecules (protein, starch, reducing sugar) in four unknown solutions (S1, S2, S3, S4) and a mixture (U) using specific reagents. It provides detailed steps for conducting tests, recording observations, and identifying the presence of these molecules. Additionally, it includes a section on testing two other solutions (Y and Z) for the same molecules and discusses potential conclusions and further testing for enzyme identification.

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alekspydie
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23 views5 pages

Biological Molecule Testing Syllabus

The document outlines an experiment to identify biological molecules (protein, starch, reducing sugar) in four unknown solutions (S1, S2, S3, S4) and a mixture (U) using specific reagents. It provides detailed steps for conducting tests, recording observations, and identifying the presence of these molecules. Additionally, it includes a section on testing two other solutions (Y and Z) for the same molecules and discusses potential conclusions and further testing for enzyme identification.

Uploaded by

alekspydie
Copyright
© All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd

EXPERIMENT NO.

1
Before you proceed, read carefully through the whole of Question 1 and Question 2.

Plan the use of the two hours to make sure that you finish the whole of Question 1 and Question 2.

1 Some biological molecules produce a colour change when tested with a specific reagent.

You will investigate which biological molecules are present in four solutions, S1, S2, S3, and S4.
Each solution contains one or none of the biological molecules: protein, starch, reducing sugar.

You will also investigate the contents of solution U which contains a mixture of biological molecules.

You are provided with the materials shown in Table 1.1.

Table 1.1

labelled contents hazard volume


/ cm3
C 0.15% copper sulfate none 10
solution
P 5% potassium hydroxide harmful 10
solution irritant
iodine iodine solution irritant 5

B Benedict’s solution harmful 20


irritant
S1 unknown solution none 10

S2 unknown solution none 10

S3 unknown solution none 10

S4 unknown solution none 10

U unknown mixture of none 10


biological molecules

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

It is recommended that you wear suitable eye protection.

9700/33/M/J/21
Carry out the test for protein on S1, S2, S3, and S4, using step 1 to step 9.

1. Label four test-tubes, S1, S2, S3, and S4.

2. Put 1 cm3 of S1 into the test-tube labelled S1.

3. Repeat step 2 for S2, S3, and S4.

4. Put 1 cm3 of P into each test-tube. Shake gently to mix.

5. Put 1 cm3 of C into each test-tube. Shake gently to mix.

6. Leave the test-tubes for at least 1 minute. Shake gently to mix.

7. Observe the colour in each test-tube and record your observations in (a)(iii).

8. Identify the solution or solutions (S1, S2, S3, S4) that contain protein and record this in (a)(iv).

9. Keep any test-tubes from step 8 that contain protein. Empty the test-tubes that do not contain
protein into the container labelled ‘For waste’. Rinse the empty test-tubes for use in step 10.

(a) (i) State the reagent that is used to test for reducing sugar and the colour or colours
produced if a reducing sugar is present.

reagent ..............................................

colour or colours ................................................................................................................

Describe how you will use the reagent to carry out the test for reducing sugar.

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

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

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

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

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

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

[3]

10. Carry out the test for reducing sugar on each of the solutions using the labelled test-tubes
from step 9. You do not need to test any solutions identified as containing protein in step 8.

11. Observe the colour in each test-tube and record your observations in (a)(iii).

12. Identify the solution or solutions (S1, S2, S3, S4) that contain reducing sugar and record this
in (a)(iv).
13. Keep any test-tubes from step 12 that contain reducing sugar. Empty the test-tubes that do
not contain reducing sugar into the container labelled ‘For waste’. Rinse the empty test-tubes
for use in step 14.

(ii) State the reagent that is used to test for starch and the colour that is produced if starch is
present.

reagent ..........................................................

colour ............................................................

[1]

14. Carry out the test for starch on each of the solutions using the labelled test-tubes from
step 13. You do not need to test any solutions identified as containing protein or reducing
sugar in step 8 and step 12.

15. Observe the colour in each test-tube and record your observations in (a)(iii).

16. Identify the solution or solutions (S1, S2, S3, S4) that contain starch and record this in (a)(iv).

17. Identify the solution or solutions, if any, that do not contain protein, reducing sugar or starch.
Record this as ‘none’ in (a)(iv).

(iii) Record your observations in an appropriate table.

[4]
(iv) Identify the biological molecules present in solutions S1, S2, S3 and S4. If any solution
does not contain protein, reducing sugar or starch write the answer as 'none'.

S1 ......................................................................................................................................

S2 ......................................................................................................................................

S3 ......................................................................................................................................

S4 ......................................................................................................................................
[3]

18. Carry out the tests for protein, reducing sugar and starch using solution U.

19. Observe the colour from each test and record your observations in (a)(v).

(v) Record the colours observed for each test for U.

protein test ............................................................

reducing sugar test ................................................

starch test ..............................................................


[1]

(vi) Identify the biological molecules present in U.

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

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

...........................................................................................................................................
[1]
(b) A student carries out the same tests for protein, reducing sugar and starch on two unknown
solutions, Y and Z. The student also carries out the test for the presence of lipid.

(i) Describe the test for lipid and the expected observation if a lipid is present.

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

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

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

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

observation if a lipid is present ..........................................................................................


[2]

From the observations for each test, the student concludes:

• solution Y contains protein and reducing sugar


• solution Z contains protein and lipid.

The student also carries out the test for non-reducing sugar on solution Y and on solution Z.

The student observes a positive result for the presence of a non-reducing sugar in solution Y
and in solution Z.

The student concludes that a non-reducing sugar is present in solution Y and in solution Z.

(ii) Suggest why the student’s conclusion may not be correct for solution Y.

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

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

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

(iii) The student thought that the protein in solution Z could be the enzyme amylase.

Describe how the student could determine whether the protein in solution Z is amylase.

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

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

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

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

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

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

...........................................................................................................................................
[3]
[Total: 19]

Common questions

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To confirm the presence of proteins in a solution, add 1 cm³ of the solution to a test-tube, then add 1 cm³ of 5% potassium hydroxide solution and 1 cm³ of 0.15% copper sulfate solution. Shake gently and observe the color after one minute. A color change indicating the presence of proteins is typically a violet or purple hue .

It is important to wear suitable eye protection to prevent irritation from reagents like potassium hydroxide, iodine, and Benedict’s solution, which are harmful irritants. Additionally, one should ensure that if any solution comes into contact with the skin, it should be washed off immediately with cold water to minimize harm. Proper lab hygiene requires rinsing test-tubes after use and disposing of waste solutions in labelled containers .

A systematic approach involves sequential testing: first testing for proteins using potassium hydroxide and copper sulfate, then for reducing sugars using Benedict’s solution, followed by starch testing with iodine. Each test should be observed for expected color changes corresponding to the presence of specific molecules. Maintaining a clear sequence and documentation helps in accurately identifying the molecules .

To test for reducing sugars, add Benedict's solution to the mixture and heat it in a boiling water bath. A positive result will be indicated by a color change ranging from green to brick-red, depending on the amount of reducing sugar present. The solution should be warmed for a sufficient duration to observe this color change .

A lipid test involves using a simple procedure like the emulsion test, where the sample is mixed with ethanol, followed by water. A milky-white emulsion indicates the presence of lipids. This result is due to the dispersion of lipid droplets in the aqueous phase, differentiating it from protein, starch, or sugars .

Ensure comprehensive recording by using clear and detailed tables with columns for each test and observations. Record exact color changes and any anomalies. For evaluation, compare expected outcomes with actual results, and re-test if necessary to verify inconsistencies or unexpected findings. Thorough documentation includes all procedural notes and any deviations from standard methods .

The student's conclusion might be incorrect if the test for non-reducing sugars was not performed correctly or if there were interfering substances affecting the test results. Additionally, incomplete hydrolysis or failure to neutralize the acid could result in false negatives or misidentification .

To determine if amylase is present, perform an enzymatic activity assay where the solution is mixed with a starch substrate and incubated at optimal temperature and pH. Measure the breakdown of starch by periodically testing for reducing sugars using iodine or Benedict's reagent. A decrease in starch and presence of maltose confirm amylase activity .

A positive test for both reducing and non-reducing sugars could occur if the solution contains both types of sugars separately or if a disaccharide that is initially non-reducing was hydrolyzed to smaller reducing sugars. To confirm, perform hydrolysis by heating with acid or using specific enzymes such as invertase, then proceed to test with Benedict's solution. This confirms the presence of additional reducing sugars post-hydrolysis .

To test for starch, add iodine solution to the sample. The presence of starch will result in a blue-black color change. This reaction occurs as the iodine interacts with the coiled structure of the starch molecule .

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