Food tests

1. General guidelines
Test samples can either be a:
Solid sample:
o Organic molecules that you are testing 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.
Liquid sample:
o Organic molecules will already be dissolved/ suspended in the liquid, hence
solvents are not required in this case.
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 (Benedicts Test) [Only test requiring
heating!]
Examples of reducing sugars include monosaccharides and disaccharides (but
excluding sucrose): Glucose, galactose, fructose, maltose & lactose.
These sugars react with copper (II) ions (blue) in Benedicts solution and cause
these ions to be reduced to copper (II) oxide (red), which is insoluble (it comes
out as a brick-red precipitate).
For liquid test solution (use 2 cm3):
1. Add equal volume of Benedict 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 (ex: groundnut/ fruit sample):
1. Crush part of the solid food sample, then add it to a test-tube which is 5 cm3
filled with water (or one quarter of the test tube). Shake thoroughly to mix the
sample.
2. Pour half the contents of this test tube into another, empty test tube and test it
for reducing sugar. Follow the procedure for liquid sample.
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 & Benedicts
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 waterbath, and record down colour changes. The solution with the highest
concentration of reducing sugar will show a colour change first.
Note: Sometimes, when it seems like there is not colour change or no precipitate
forms, place the test tube in the test tube rack first and make a second

observation 5-10mins later (to allow precipitate to be formed, and to settle at the
bottom of the test tubes).
Observations:
Amount of Reducing Sugar
Nil/ Absent
Small amount
Moderate amount
Large amount

Appearance of contents of test tube

Benedicts solution remains blue
Green precipitate is formed
Yellow precipitate is formed
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.

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 (ex: groundnut/fruit):
1. Crush the solid food sample and place it on a petri dish.
2. Use a dripper to add iodine solution to it. Ensure the food is immersed in the
iodine solution.
3. After 5-10mins 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 ad the iodine
solution directly to the sample.
Observations:
Presence of starch:
For liquid test solution:
The yellow iodine solution turned the test solution 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.
Absence of starch:
The iodine solution/ food particles remain yellow in colour.

3. Test for Proteins (Biuret Test)

This test involves the reaction of copper (II) ions (in copper (II) sulphate solution) which
give a blue colour, with the peptide bonds that hold the amino acids together, producing
a violet colouration. This reaction requires and alkaline medium, hence sodium
hydroxide is added first.
For solid food (ex: groundnut):
1. Crush the food and prepare a solution as described in the Benedicts Test.

2. Add equal volume of sodium hydroxide solution to the crushed food solution
(about 2 cm3). Shake GENTLY.
3. Add the 1% copper (II) sulphate solution provided, drop by drop, shaking
GENTLY after every drop, to the mixture. Observe the colour change.
If the result above 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 cm3
above its level). Shake GENTLY.
3. Add the 1% copper (II) 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 cm3 of water to a test-tube, ad adding an equal
volume of sodium hydroxide.
2. Add the same number of drops of 1% copper (II) sulphate 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 (ex: ethanol) is used to extract out the
lipids.
For liquid test solution:
1. Add 5 cm3 of alcohol (or fill a test tube one quarter full) 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 (ex: groundnut):
1. Crush part of the solid food sample 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)
Absence of lipids: Alcohol remained clear/ colourless.