FFP Lab Manual 22FTBS152

EXPERIMENT NO- 1

AIM: Estimation of reducing sugar by Fehling’s procedure.

PRINCIPLE:

One of the most popular tests used for the estimation or detection of reducing sugars and non-reducing sugars
is the Fehling’s test. The test developed by German chemist H.C. Von Fehling is also used to differentiate
between ketone functional groups and water-soluble carbohydrates. Fehling’s test consists of a solution that is
usually prepared fresh in laboratories. Initially, the solution exists in the form of two separate solutions which
are labelled as Fehling’s A and Fehling’s B. Fehling’s A is a solution containing copper(II) sulphate, which is
blue. Fehling’s B is a clear liquid consisting of potassium sodium tartrate (Rochelle salt) and a strong alkali,
usually sodium hydroxide. During the test solutions A and B are prepared individually and stored. The two
solutions are later mixed in equal volumes to get the final Fehling solution which is deep blue. The deep blue
ingredient is the bis(tartrate) complex of Cu2+. The tartrate tetra-anions serve as a chelating agent in the
solution.

The reducing ability of sugars can be determined by their ability to reduce cupric ions in alkaline solution to
form red cuprous oxide. The best-known reagent for this test is Fehling’s solution. A positive reaction is
shown by a green, yellowish-orange, or red coloration of precipitate, depending upon the amount and kind of
reducing sugars. The degree of color change from blue to red is an indication of the extent of the reduction
and therefore a measure of the reducing power of the sugar.

MATERAIL METHODS: Fehling’s solution, Water, Sample (juice), Test tubes.

REACTIONS:

REAGENTS:

a) Fehling’s solution (A+B): Fehling's "A" uses 7 g CuSO4.5H2O dissolved in distilled water
containing 2 drops of dilute sulfuric acid.
b) Fehling's "B" uses 35g of potassium tartrate and 12g of NaOH in 100 ml of distilled water.
FFP Lab Manual 22FTBS152

PROCEDURE:

1. Add the sample (1 mL) in a test tube.

2. Add distilled water in another test tube for control.
3. Add 5 mL of Fehling’s solution in both the test tube.
4. Keep the test tubes in water bath.
5. Make observation and record if there is any development of red precipitate.

Note: Notably, the result is positive if there is a formation of reddish brown precipitate while
the result is negative if there is no indication of such change.

PRECAUTIONS

1. Weigh the chemicals accurately.

2. Make the solutions carefully