Benedict's Test for Non-reducing Sugars

The Benedict's Test for Non-reducing Sugars determines the presence of non-reducing
sugars - sugars which do not have an aldehyde functional group.
Many students confuse the Benedict's Test for Non-Reducing Sugars with the Benedict's
Test for Reducing Sugars, perhaps because some of the steps in the procedure are similar.
Benedict's solution is the principle reagent in both tests. The test for non-reducing sugars is
often conducted on a food sample which tested negative for reducing sugar. If reducing
sugars have been shown to be present, a heavier precipitate is often observed when the test
for non-reducing sugar is conducted.
The Fehling's Test for Non-reducing sugar is an alternative to the Benedict's Test. However it
is less popular as it less sensitive and requires tha the reagents - Fehling's solutions A and B be kept separate until the experiment is carried out.

What is a Non-Reducing Sugar?

Sugars can be classified as either reducing or non-reducing based on their ability to reduce
copper(II) ions to copper (I) ions during the Benedict's Test.
Non-reducing sugars do not contain an aldehyde group - the reducing species.
Reducing sugars are simple, disaccharide sugars. Sucrose is the most common disaccharide
non-reducing sugar.

Procedure
1.A liquid food sample does not need prior preparation except dilution if viscous or
concentrated.
For a solid sample prepare a test solution by crushing the food and adding a moderate
amount of distilled water. Decant the suspension to remove large particles. Use the
decanted liquid as the test solution.
2.Add 2 cm3 of the sample solution to a test tube.
3.Add 1 cm3 of dilute hydrochloric acid and boil for one minute.
4.Allow the tube to cool and then neutralize the acid with sodium hydrogen
carbonate. Exercise caution due to 'fizzing' or effervescence.
Check with blue litmus paper or pH paper. Do not worry if the resulting solution is
slightly alkaline; Benedict's solution is also alkaline since an alkaline medium in
needed for oxidation of the copper (II) ions by the sugars
Then carry out Benedict's Test as for reducing sugars:
5.Add an 2 cm3 of Benedict's solution to the test tube and swirl or vortex the
mixture.
6.Leave the test tube in a boiling water bath for about 5 minutes, or until the colour
of the mixture does not change.
7.Observe the colour changes during that time as well as the final colour.
8.To prepare a control, repeat steps 2-6 using 2 cm3 of distilled water instead of
sample solution.

Observations and Interpretation

OBSERVATION
(FINAL COLOUR CHANGE)

INTERPRETATION

No colour change (mixture remains blue)

No non-reducing sugar present

Green

Trace amounts of non-reducing sugar present

Yellow

Low amounts of non-reducing sugar present

Orange

Moderate amounts of non-reducing sugar present

Brick Red

Large amounts of non-reducing sugar present

Whilst boiling the colours of the mixture transition in this order:

blue--> green--> yellow--> orange--> brick red

The final colour may be any of the colours above, depending on the quantity of non-reducing
sugar present. If you do no observe the brick-red colour, it does not mean that your
experiment has not been successful.
It is important to note that the Benedict's Test for Non-reducing Sugars is not specific to any
one type of non-reducing sugar, and that the colour corresponds to the total
reducing and non-reducing sugar present.

Principle of the Benedict's Test for Non-reducing Sugar

Disaccharides are hydrolyzed to their constituent monosaccharides when boiled in dilute
hydrochloric acid. The monosaccharides are reducing sugars are they contain the aldehyde
group which is the reducing species.
Neutralization of the mixture by sodium hydrogen carbonate is necessary as the reduction
of the copper(II) ions will not take place in acidic conditions - of excess acid is present.
Benedict's Solution contains copper(II) sulphate, sodium carbonate and sodium citrate.
The blue copper(II) ions from copper(II) sulphate are reduced to red copper(I) ions by
the aldehyde groups in the reducing sugars. This accounts for the colour changes observed.
The red copper(I) oxide formed is insoluble in water and is precipitated out of solution. This
accounts for the precipitate formed. As the concentration of reducing sugar increases, the
nearer the final colour is to brick-red and the greater the precipitate formed.
Sodium carbonate provides the alkaline conditions which are required for the redox reaction
above.
Sodium citrate complexes with the copper (II) ions so that they do not deteriorate to
copper(I) ions during storage.