Benedict's Test for Reducing Sugars

The Benedict's Test for Reducing Sugars is used to determine the presence of reducing sugars. There is a Benedict's Test for non-reducing sugars as well. Benedict's solution is the principle reagent in the Benedict's Tests. The Fehling's Tests for reducing as well as for non-reducing sugar are alternatives to the Benedict's Test. Although based on the same principle, the Fehling's Test is less sensitive. It is also less convenient as the Fehling's Reagents - Fehling's A and B - have to be kept separate until the test is conducted.

What is a Reducing Sugar?

Sugars are classified as reducing or non-reducing based on their ability to act as a reducing agent during the Benedict's Test. A reducing agent donates electrons during a redox reaction and is itself oxidized. The aldehyde functional group is the reducing agent in reducing sugars. Reducing sugars have either an aldehyde functional group or have a ketone group - in an open chain form which can be converted into an aldehyde. Reducing sugars are simple sugars and include all monosaccharides and most disaccarides. Some examples of monosaccharides are glucose, fructose and galactose.Examples of reducing disaccharides are lactose and maltose. Note that the disaccharide sucrose is not a reducing sugar. In fact, sucrose is the most common 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 an equal volume of Benedict's solution to the test tube and swirl or vortex the mixture. 4.Leave the test tube in a boiling water bath for about 5 minutes, or until the colour of the mixture does not change. 5.Observe the colour changes during that time as well as the final colour. 6.To prepare a control, repeat steps 3-5 using 2 cm3 of distilled water instead of sample solution.

Observations and Interpretation

OBSERVATION (FINAL COLOUR CHANGE)
No colour change (mixture remains blue) Green Yellow Orange Brick Red

INTERPRETATION
No reducing sugar present Trace amounts of reducing sugar present Low amounts of reducing sugar present Moderate amounts of reducing sugar present Large amounts of reducing sugar present

During the experiment, the colours of the mixture transition in this order: blue--> green--> yellow--> orange--> brick red

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

Principle of the Benedict's Test for Reducing Sugars

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.