To2.3 Topic 2.

3
Carbohydrates disaccharides and
Practical worksheet polysaccharides

Food test 2: biochemical tests for

non-reducing sugars and starch
Aims
In this practical work you will carry out two qualitative food tests to find out whether certain
types of food molecules occur in various food samples. For example, does starch occur in
potatoes?
You will test materials that should give positive results and test controls that should give
negative results.
You can practise manual, observational and data handling skills and then go on to make
conclusions.
There are opportunities for you to evaluate information in the discussion sections.

The test for non-reducing sugars using

Benedicts reagent after acid hydrolysis
Some disaccharides are non-reducing sugars. This is because the monosaccharide units
within the disaccharide that could potentially cause the reduction reaction are involved in the
glycosidic bond between the two monosaccharide units. (The chemical groups are aldehyde
or ketone groups.)

Safety
Wear eye protection there is some risk of hot liquids spitting when heating test tubes
Benedicts reagent: LOW HAZARD
Dilute hydrochloric acid (0.1 mol dm3): LOW HAZARD but may cause harm if it enters
the eye or a cut
Sodium hydrogencarbonate: MINIMAL HAZARD IN THIS TEST
Take care when cutting food with a scalpel
LET YOUR TUTOR KNOW IF YOU HAVE A NUT ALLERGY. You should not carry
out tests with nuts if you are allergic to them.

Equipment and materials

As for the test for reducing sugars (see food test 1) plus three items:
Dilute hydrochloric acid (0.1 mol dm3)
Sodium hydrogencarbonate powder
Universal indicator paper and chart

Method
1. Proceed with this test only after a negative Benedicts test for reducing sugars on the
food.

Oxford University Press 2014 http://www.oxfordsecondary.co.uk/acknowledgements

This resource sheet may have been changed from the original.
1
 To2.3 Topic 2.3
Carbohydrates disaccharides and
Practical worksheet polysaccharides

2. Half fill a 400 cm3 beaker with tap water to act as a water bath. Use a heatproof mat,
tripod, gauze and Bunsen burner to heat the water to boiling point.
3. Label the tops of your test tubes.
4. Use a Pasteur pipette to dispense 2 cm3 of a fresh sample into a test tube (or finely
chopped/ground food to 1 cm depth and 2 cm3 of distilled water).
5. Use a dropper pipette to add 10 drops of dilute hydrochloric acid (HCl) to the sample.
6. Use a test tube holder to transfer the test solution to the water bath. You can save time by
having up to five test tubes in the water bath. Heat to boiling point for two minutes. This
is the stage when acid hydrolysis may occur as discussed below.
7. Cool the test tube in cold water (in a beaker or under the cold tap).
8. Sodium hydrogencarbonate (NaHCO3) powder is needed to make the solution alkaline.
With a spatula add sodium hydrogencarbonate to the solution until the fizzing stops. The
solution must be alkaline for the Benedicts reagent to work. You can confirm that the
solution is alkaline by using a Pasteur pipette to take a sample and adding it to universal
indicator paper on a tile; compare the colour with that in the chart.
9. Add 10 drops of Benedict's reagent, about 0.5 cm3, to the test solution.
10. Heat the test solution to boiling point in a water bath and continue heating for two
minutes.
11. Observe the colour changes in the test tubes over two minutes of heating at boiling point.
Record your observation in Table 1.

Results
Confirmation of a non-reducing sugar, for example, sucrose, in the original solution: a
brick-red/brown/orange/yellow or green suspension at this stage only.
Confirmation of the absence of a non-reducing sugar (and the absence of reducing
sugars) in the original solution: a blue solution remains.

Table 1 Observations and deductions from the test for non-reducing sugars using Benedicts
reagent

Oxford University Press 2014 http://www.oxfordsecondary.co.uk/acknowledgements

This resource sheet may have been changed from the original.
2
 To2.3 Topic 2.3
Carbohydrates disaccharides and
Practical worksheet polysaccharides

Material tested Observations (after acid and heat, Deductions

etc.)
1% sucrose solution

Distilled water (control)

1% starch suspension

The explanation of the test for non-reducing sugars using

Benedicts reagent
Sucrose is a non-reducing sugar that occurs naturally in plants. Sucrose is a disaccharide
consisting of glucose and fructose units linked by a glycosidic bond. It is not a reducing sugar
because the reducing groups of glucose and fructose are tied up in the glycosidic bond. When
a sucrose solution is acidified and heated; the hydrogen ions of the acid catalyse the
hydrolysis of sucrose to glucose and fructose, which are both reducing sugars, and cause the
reduction reaction.

Question
1. Which of the materials you tested:
a) contained reducing sugars? ...

...

b) contained non-reducing sugars?

...

c) contained neither reducing sugars nor non-reducing sugars? ...

...

Discussion
1. A student did not read the practical schedule carefully enough and found a brick-red
suspension for the test for reducing sugars and the test for non-reducing sugars.
a) What can be concluded from these results?

.......................................................................................................................................
Oxford University Press 2014 http://www.oxfordsecondary.co.uk/acknowledgements
This resource sheet may have been changed from the original.
3
 To2.3 Topic 2.3
Carbohydrates disaccharides and
Practical worksheet polysaccharides

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

b) What cannot be concluded from these results?

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

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

2. Trehalose is also a non-reducing sugar formed from two glucose units that are joined by a
1-1 alpha glycosidic bond (unlike the 1-4 alpha glycosidic bond between the two glucose
units of the reducing disaccharide, maltose). The glycosidic bond of trehalose is very
difficult to hydrolyse.
Suggest why trehalose might not give a positive result in the test for non-reducing sugars
using Benedicts reagent.

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

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

The iodine test for starch

Safety
Wear eye protection
Iodine solution avoid contact with skin and clothes
Take care when cutting food with a scalpel

Equipment and materials

Eye protection Spatula
Labelling equipment Distilled water
Scalpel and tile Iodine (in potassium iodide)
solution
Pasteur pipettes
1% starch suspension
Dimple tile
Food materials, for example, potato
and onion

Method
1. Label individual dimples on the dimple tile with the foods to be tested.
2. Use a clean Pasteur pipette to dispense a few drops of the test solution into a labelled
dimple. For solid foods, use the scalpel to cut the food sample into small pieces on a tile
and use a clean spatula to transfer some of them into a labelled dimple.
3. Use the dropper pipette to add two drops of iodine solution (iodine in potassium iodide)
to the food samples.

Oxford University Press 2014 http://www.oxfordsecondary.co.uk/acknowledgements

This resource sheet may have been changed from the original.
4
 To2.3 Topic 2.3
Carbohydrates disaccharides and
Practical worksheet polysaccharides

Results
Confirmation of starch: a blue-black coloration
Confirmation of the absence of starch: a yellow iodine colour (or just no blue-black
colour)
Table 2 Observations and deductions from the iodine test for starch

Material tested Observations Deductions

1% starch suspension

The explanation of the iodine test for starch

Starch contains both amylose and amylopectin molecules. The coils of its amylose molecules
take up iodine molecules from the iodine solution and form the blue-black, starchiodine
(polyiodide) complex.

Discussion
1. Which of the materials you tested
a) contained starch?
.......................................................................................................................................

b) did not contain starch?

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

2. How could you show that a yellow liquid contains iodine?

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

Oxford University Press 2014 http://www.oxfordsecondary.co.uk/acknowledgements

This resource sheet may have been changed from the original.
5