VLD 421: Veterinary Clinical Biochemistry and Laboratory Diagnosis-II (0+1)

Estimation of Glucose

METHOD OF ESTIMATION OF GLUCOSE - GOD-PAP METHOD

Sample

Method of Estimation : GOD-PAP Method

Principle

• Enzymatic colorimetric determination of glucose according to the following reaction.

Glucose +O2 ^{Glucose Oxide} Gluconic acid + H₂O₂

2H₂O₂ + Phenol + 4 – Aminoantipyrine ^Peroxidase Red quinone + 4H2O

Reagent composition

• Glucose R1
  • Phosphate buffer, (pH 7.40) 100 mmol/L
  • Phenol 10 mmol/L
• Glucose R2
  • Glucose Oxidase >10000 U/L
  • Peroxidase > 600 U/L
  • 4-Aminoantipyrine 270mmol/L

Laboratory procedure

	Blank	Standard	Sample
Reagent	1000μL	1000μL	1000μL
Standard	--	10 μL	--
Sample	--	--	10μL
Mix and incubate for 10 min. at 37 ° C. Measure the absorbance of standard and sample against reagent blank.

Calculation

• Glucose Conc. (mg /dl) = (Absorbance of sample /Absorbance of standard) x 100
• Glucose standard concentration = 100mg/dL