o- toluidine method

this method is simple, rapid and precise.

it can be directly applied to serum or plasma without deproteinization

values obtained are nearer to true glucose

But, this method is no longer widely used because o-toluidine is

believed to be carcinogen.

Principle- Glucose present in the filtrate reacts with o-toluidine in

glacial acetic acid to form a blue green compound, namely, N-
glucosylamine.

The intensity of the colour developed is proportional to the conc

of glucose present in the blood. This is compared with a similarly
treated solution of standard glucose. The OD readings are recorded at
650nm.

Reagents-

1. o- toluidine reagent
2. trichloroacetic acid
3. Stock standard glucose solution
4. Working Standard

Procedure

Reagent Blank Standard Test

O-toluidine 5ml 5ml 5ml
Serum/plasma - - 0.1ml
dil. glucose - 0.1ml -
standard
D/W 0.1ml - -

mix thoroughly.

cover tubes with aluminium foli and place in a boiling water bath for
10min.

Cool under a tap water and note reading in photometer at 625nm (red
filter).