Basic Concepts in Clinical Biochemistry-A Practical Guide.7z

15.3 Principle........................................

The estimation of plasma glucose is based on the principle as explained for glucose
measurement by glucose oxidase method.

15.4 Reagents........................................

1.Phosphate buffer (0.1 M, pH 7.0).
2.Enzyme reagent: Prepare as explained in previous experiment.
3.Stock glucose solution (500 mg/dl): Dissolve 500 mg glucose in 100 ml distilled
water.
4.Working glucose standard: Prepare glucose standards of concentration 50, 100,
150, 200, and 250 mg/dl.

17.4 Procedure

1. Take set of 11 test tubes in duplicate and label as blank, standard (S 1 – S 5 ), and test
   samples (T 1 – T 5 ).


2. Add 1.8 ml phosphate buffer to all the test tubes. In blank add 1.9 ml phosphate
   buffer.


3. Add 1 ml enzyme reagent to all the test tubes.


4. Add 0.1 ml of standards of concentration 50, 100, 150, 200, and 250 mg/dl to the
   tubes labelled as S 1 – S 5 , respectively.


5. Add 0.1 ml of serum sample to test tubes (T 1 – T 5 ). Mix the content of the tubes
   thoroughly, and place them in a water bath at 37C for 5 min or at room
   temperature for 15 min.


6. Measure the OD of the test and the standards against blank at 520 nm.

64 15 To Perform Glucose Tolerance Test