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Enzymatic method yields maximum specificity for glucose estimation. Glucose can be measured by its reaction with glucose oxidase, in which gluconic acid and hydrogen peroxide are formed. Hydrogen peroxide than reacts with an oxygen acceptor, such as ortho-dianisidine,phenylamine-phenazone or any other chromogenic oxygen acceptors, in a reaction catalysed by peroxidase to form a colour.
One of the advantages of enzymatic method is its specificity. This enables the estimation of glucose even in a complex mixture. Thus, this method is widely used in the field of clinical chemistry and for food analysis. In clinical chemistry, the enzymatic analysis of glucose in blood and urine has been modified as dipstick test. However in the present practical, the conventional enzymatic method shall be used.

Advantages of Glucose Oxidase Method:
1. This method most specific enzymatic than other method such as; reduction of cupric to cuprous salts, and reduction of ferricyanide to ferrocyanide method.
2. The reaction can be monitored polarographically either by measuring the rate of
disappearance of oxygen.