ORTHOTOLUIDINE METHOD FOR BLOOD GLUCOSE DETER MINATION

MANUAL PROCEDURE:

Precautions:

1. Patient should have fasted for 8-12 hours.
2. Unhemoylzed, non icteric and non turbid serum should be used to avoid interferences
with the procedure.
3. The reagent contains sodium azide, which is corrosive, so PPE should be worn.
4. The solution should be brought to a boil to ensure proper reaction.

PROCEDURE PROPER:

1. Prepare Three tubes labeled, SA (Sample)/ C (Control), ST (Standard) and RB (Reagent Blank).

2. To the SA/C tube add 0.1 ml of the test specimen/ Control specimen.

3. To the ST tube add 0.1 ml of the standard stock solution.

4. To the RB tube add 0.1 ml of distilled water.

5. Add 4 ml of glucose reagent to all tubes. Cap and mix by gentle inversion.

6. Boil all tubes at 100 degrees centigrade for 8 minutes.

7. Cool tubes in running tap water for about 2 minutes.

8. Read absorbance of solutions in the spectrophotometer with appropriate cuvets at 636 nanometers against the RB (Reagent Blank).

9. Compute for the Cu (Concentration of the Unknown making use of the formula:

Cu= Absorbance of Unknown(Au) multiplied by the Concentration of the
Standard(Cs) divided by the Absorbance of the Standard (As)

Adapt the unit of the Cs

Normal Values = 60 - 100 mg/dL
to convert values to mmol/L multiply values in mg/dL with 0.0555