1. Determination of Blood Glucose By Alkaline Copper Reduction Method
LAB # 2

2. Determination of Blood Glucose (BG)
2- Alkaline copper reduction method (Asatoor & King method):
Principle
Glucose + cupric cuprous + gluconic acid
Cuprous + phosphomolybdic acid cupric + molybdenum blue

3. Determination of Blood Glucose (BG)
Procedure
1- Deproteinization
1-Add 0.1 ml
Of Na tungstate
3- add
0.1 ml
serum
2- add 3.8 ml of
Isotonic
Na2SO4/CuSO4
Soln.
-Mix well
-Label
centrifuge
Clean &
Dry
Cent. tube

4. Procedure: cont. Transfer clear supernatant Label as to clean dry t.t
Using
Pasteur pipette
Label as
supernatant
Test tube

5. Procedure: cont. Test Standard Blank T B S Add 1 ml of supernatant
+1 ml of alkaline
tararate
Put in boiling water bath for 10 mins
Mix well and close them with cotton wool
Add 1ml
Standard
Glucose
+1 ml alkaline
tararate
Cool under tap water
Add 3 ml of phosphomolybdic reagent to each
tube
Shake well
1 ml isotonic
Na2SO4/CuSO4
Soln. +
1 ml alkaline
tararate
Mix well to remove all air bubbles
Let the tubes stand for 5 mins.
Measure the absorbance of test and stand. against blank at
680 nm
Add 3 ml of distilled water to each tube

6. Calculation Glucose conc. (mg/dl) = Test abs. X Stand. conc.
Stand. abs.