DNSA CALORIMETRIC METHOD

PRINCIPLE:

This tests for the presence of free carbonyl group. It involves oxidation of the aldehyde
functional group. 3,5-dinitrosalicylic acid is reduced to 3-amino, 5-nitrosalicylic acid under
alkaline conditions. Dissolved oxygen can interfere with glucose oxidation. Sulfite is added to
absorb the dissolved oxygen. One mole of sugar will react with one mole of 3,5-dinitrosalicylic
acid.

REAGENTS:

Dinitrosalicylic acid reagent solution, 1%

Dinitrosalicylic acid: 10g

Sodium sulfite 0.5g

Sodium hydroxide 10g

Water 1 litre

Potassium sodium tartrate solution 40%

PROCEDURE:

Add 3ml of DNS reagent to 3ml of glucose sample in a test tube.

Heat the mixture at 90 C for 5-15 minute to develop red-brown colour.

Add 1ml of a 40% potassium sodium tartrate solution to stabilize the colour.

After cooling to room temperature in a cold water bath, record the absorbance with a
spectrophotometer at 570nm

Run standards with glucose along with each sets.