Testing for biological molecules

Sugars

The test for reducing sugars

Method
1. Set up five test tubes containing equal volumes of distilled water, 0.1%
glucose solution, 1.0% glucose solution, 10% glucose solution and
sucrose solution.
2. Add Benedict’s solution to each tube and heat in a water bath.

Results

Solution tested Colour at end of test

A Distilled water Blue
B 0.1% glucose Green
C 1.0% glucose Orange
D 10% glucose Red
E sucrose

Conclusion
1. What does Benedict’s solution contain and what colour is it?
Benedict’s is a deep blue colour and it contains an alkaline copper(II)
sulfate

2. Describe and explain the reaction of Benedict’s solution with a reducing

sugar.
Benedict’s solution contains Cu 2+ ions which are reduced to Cu+ ions
forming orange/red copper(I) oxide

3. Which of your solutions contained a reducing sugar?

B,C,D

4. Explain the differences in the final colour observed in test tubes B, C

and D.
B has a lower solution of glucose therefore the colour is weaker as only
a little bit of precipitate is formed, the colour of C is more stronger as
there is a stronger glucose solution so forms a darker precipitate and D
is the darkest as the colour is proportional to the concentration of sugar
so as the concentration is high more sugar is present.

The test for non-reducing sugars

Method
1. Take a fresh sample of sucrose solution in a boiling tube and add a few
drops of hydrochloric acid.
2. Boil gently.
3. Add sodium hydrogen carbonate powder until the solution stops fizzing.
4. Test for reducing sugar as above.
 Conclusions
1. Describe and explain the colour change observed during the Benedict’s
test.
A positive result (green-yellow-orange-red) indicates that non-reducing
sugar (e.g. sucrose) was present in the original sample.

2. What was the purpose of heating the sucrose solution with hydrochloric
acid?
You have to hydrolyse the bond to free up the reducing groups

3. Why was the solution neutralised before being tested with Benedict’s
solution?
Hydrogen carbonate is then added to neutralise the solution as
Benedick's reagent doesn't work under acidic conditions.

Starch

Method
1. Set up four test tubes containing equal volumes of distilled water,
0.01%, 0.1% and 1% starch solution.
2. Add a few drops of iodine in potassium iodide solution to each tube.

Results

Solution tested Colour at end of test

A Distilled water Yellow
B 0.01% starch solution Dark blue
C 0.1% starch solution Blue/black
D 1% starch solution Black

Conclusions
1. Why was the iodine dissolved in potassium iodide solution and not in
water?
The iodine forms a triiodide ion I3- which slips into the middle of the
amylose helix

2. Explain the results of your test with reference to the shape of starch
molecules.
Longer chains yield a blue black result with iodine whereas shorter
chains give no colour change or a much less intense red purple colour

Lipids
Method
1. Pour some cooking oil into a test tube, add 1cm 3 of absolute ethanol
and shake well. Allow to settle.
2. Half fill a test tube with water and pour the ethanol from the first tube
into the water.
3. Repeat the test on distilled water.

Conclusions
1. Describe and explain your results.
A cloudy white emulsion indicates the presence of lipids. When the test is
repeated with water no emulsion will be shown.

Proteins

Method
1. Add a few drops of Biuret reagent to two test tubes, one containing
protein solution and the other containing distilled water.
2. Warm gently

Conclusions
1. What does Biuret reagent contain?
It contains sodium hydroxide and copper sulfate

2. Describe and explain your results.

If protein is present the colour changes from light blue to lilac. The
colour formed by a complex between the nitrogen atoms in a peptide
chain and Cu 2+ ions, which is why this test detects the presence of
peptide bonds.

3. In each of the experiments what was the purpose of carrying out the
test on distilled water?
Distilled water is used to check whether the experiment is accurate as
when you use distilled water the colour should not change as no
molecules were present.