Maris Stella High School Name: ___________________

GCE O Level Biology Class (Index No.): Sec 3___ ( )

Practical Guidelines 1 Date: ________________

Food Tests
1. General Guidelines
Test samples (the sample you are testing) can either be a:
• Solid sample:
o Organic molecules that you are testing for will be found inside the cells, hence
the sample must be crushed.
o After crushing, a solvent (water or alcohol) will be used to extract out the
molecules. Exception?
• Liquid sample:
o Organic molecules will already be dissolved/suspended in the liquid, hence
solvents are not required in this case. Exception?
o In all cases unless otherwise stated, the volume of test sample used would be
2 cm3.

2. Test for Carbohydrates

A. Test for Reducing Sugars (Benedict’s Test) [Only test requiring heating!]
Examples of reducing sugars include monosaccharides and dissacharides (but excluding
sucrose): Glucose, galactose, fructose, maltose & lactose.

These sugars react with copper (II) ions (blue) in Benedict’s solution and cause these ions
to be reduced to copper (I) oxide (red), which is insoluble (it comes out as a precipitate).

For liquid test solution (use 2 cm3):

1. Add equal volume of Benedict’s solution to the test solution (2 cm3).
2. Shake the mixture and heat it by placing the test-tube into a boiling water bath for
5 minutes.
3. Observe the colour changes in the mixture after 5 minutes.

For solid food (e.g. groundnut/ fruit sample):

1. Crush part of the solid food sample on the white tile provided, then add it to a test-
tube which is 5 cm3 filled with water (or one quarter of the test tube, or the amount
stated in the question). Shake thoroughly, to mix the sample.
(We use water as the solvent as reducing sugars are soluble in water.)
2. Pour half of the contents of this test-tube* into another, empty test-tube and test it
for reducing sugar. (Follow the procedure described above for liquid sample, ie, add
equal volume of Benedict’s solution, etc.)

*The remaining half of the content can be used for the protein test if the question
requires you to do so.

Comparisons:
If you are testing more than 1 sample for reducing sugar, you will need to compare
the estimated amount of reducing sugar in each sample.
To do this, you must:
1. Ensure a fair test by using a same volume of test solution & Benedict’s solution is used
for each sample: The final level in each test tube has to be the same.
2. Observe which test solution changes colour first while heating in boiling water-bath,
and record down colour changes. The solution with the highest concentration of

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O Level Biology Topic 1
reducing sugar will show a colour change first.

Note: Sometimes, when it seems like there is no colour change or no ppt formed, place the
test tube(s) in the test-tube rack first and make a second observation 5-10 mins later (to
allow ppt. if any, to settle down at the bottom of the test-tubes).

Observations:
Amount of reducing sugar Appearance of contents of test tube
Nil/Absent Benedict’s solution remains blue
Small amount Green precipitate is formed
Moderate amount Yellow precipitate is formed
Large amount Brick red precipitate is formed

**Check that you heat only for 5 minutes. A brown precipitate indicates that you have
overheated the sample, and this is not an acceptable observation.

When writing your practical report, look at the space given and marks allocated for the
question. Sometimes more details are required, and more marks may be allocated. Then
your answer can be as follows:

Reaction in this test takes place slowly, with streaks of yellow or green circulating in the
convection currents before the formation of an orange ppt.

In other words, the preliminary green or yellow ppt before the orange ppt MUST be noted.
(Taken from Examiner’s Report 1991.)

B. Test for Starch (Iodine Test)

For liquid test solution (use 2 cm3):

1. Use the dropper to add iodine solution to the test solution (2 cm3) and shake.
You MUST state the number of drops of iodine solution used.
(Note: Sometimes, one drop is already sufficient.)
2. Observe the colour change.

For solid food (e.g. groundnut/ fruit):

1. Crush the solid food sample and place it on a petri-dish.
2. Use a dropper to add iodine solution to it. Ensure the food is immersed in the
iodine solution.
3. After 5-10 minutes or longer, observe the colour change.

Note: Starch is a large molecule (polysaccharide) and hence is not very soluble in water.
Therefore, we do not add water to the solid sample, but add the iodine solution directly to
the sample.

Observations:
Presence of starch :
For liquid test solution:
The yellow iodine solution turned the test solution (e.g. solution 1B) blue-black. (NOT blue.)

For solid food sample:

The crushed food particles (not the solution) gradually turned blue-black. (NOT blue).
Note: Sometimes, some regions of the sample will turn blue-black, while others remain
brown. In such cases, you must state the parts that show a positive result, as well as the
parts that show a negative result.

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Maris Stella High School Biological Molecules
O Level Biology Topic 1
Absence of starch:
The iodine solution / food particles remained yellow in colour.

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Maris Stella High School Biological Molecules
O Level Biology Topic 1
3. Test for Proteins (Biuret Test)
This test involves the reaction of copper (II) ions (in copper sulphate solution) which give a
blue colour, with the peptide bonds that hold the amino acids together, producing a violet
colouration. This reaction requires an alkaline medium, hence sodium hydroxide is added first.

For solid food (e.g. groundnut):

1. Crush the food and prepare a solution as described in the Benedict’s Test.
2. Add equal volume of sodium hydroxide solution to the crushed food solution (about 2
cm3). Shake GENTLY.
3. Add the 1% copper sulphate solution provided, drop by drop, shaking GENTLY after
every drop, to the mixture. Observe the colour change.
If the above result is negative, try the following method:
1. Crush the solid food sample and place it in the test-tube.
2. Add sufficient sodium hydroxide solution to the crushed food sample (about 2 cm above
its level). Shake GENTLY.
3. Add the 1% copper sulphate solution, drop by drop, shaking GENTLY after every drop,
to the mixture. Observe the colour change.

For liquid test solution:

1. Add equal volume of sodium hydroxide solution to the test solution (2 cm3). Shake the
mixture GENTLY.
2. Add the 1% copper sulphate solution provided, drop by drop, shaking GENTLY after
every drop, to the mixture. Observe the colour change.

Observations:
If a lilac colour is observed, it indicates the presence of proteins.
If mixture remains blue, it indicates absence of proteins.
Note: You may need to do a blank to compare the results.
1. A blank is prepared by adding 2 of water to a test-tube, and adding an equal
volume of sodium hydroxide.
2. Add the same number of drops of 1% copper sulphate solution as you have added
to the test sample, and compare the colours of the blank and the test sample.

4. Test for Lipids (Emulsion Test) [Ethanol must be kept away from the flame!]
Lipids are not water soluble, so an organic solvent (eg ethanol) is used to extract out the lipids.

For liquid test solution:

1. Add 5 cm3 of alcohol (or fill a test tube one quarters full, or the amount specified in the
question) to the test solution (2 cm3) and leave the test-tube in the rack for 3 minutes.
2. Decant the top clear alcohol layer to a second, dry test-tube and add a few drops of
deionised water to this second dry test-tube. Observe the colour change.

For solid food (e.g. groundnut):

1. Crush part of the solid food sample (you may need more for this test, e.g. 2 peanut seeds)
on the white tile provided, and place it in a dry test-tube.
2. Add sufficient alcohol to the crushed food sample in the dry test-tube (about 2 cm above
the level of sample) . Leave the test-tube in the rack for 3 minutes.
3. Using the filter funnel and filter paper provided, filter the contents of this test-tube into a
second, dry test-tube.
4. Add a few drops of deionised water to the filtrate in this second dry test-tube. Observe the
colour change.

Observations:
Presence of lipids: A white precipitate was produced when water was added, heat was
released in the reaction. (Test-tube was warm.)

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Absence of lipids: Alcohol remained clear / colourless.

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