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Glucose oxidase (GOD) converts Glucose to gluconic acid. Hydrogen peroxide formed in this reaction, in presence of Peroxidase (POD), oxidatively couples with 4-aminoantipyrine / phenol to produce red quinonimine dye. This dye has absorbance maximum at 505 nm. (500-550 nm.). The intensity of the colour complex is directly proportional to the Glucose in specimen.
10 x 100 ml
4 x 500 ml (Code GU-3) 4
STANDARD bottle -100 mg%
STANDARD bottle - 300 mg%
WORKING Solution Container*
Empty bottle for preparation and storage of working solution)
GLUCOSE standard 100 mg% and 300 mg% are assayed against National Bureau of Standards (NBS) reference material from Washington.
ENZYME and STANDARD are to be stored at 2-8 degree C.
Diluent Reagent may be stored below 30 degree C and away from direct light. The reagents are for in vitro diagnostic use.
PREPARATION OF WORKING SOLUTION
COMPONENTS & CONCENTRATION OF WORKING SOLUTION
Phosphate Buffer pH 7.0
SPECIMEN COLLECTION & PRESERVATION
Blood should be collected in a clean dry container. Serum or plasma should be separated from the cells at the earliest possible (within 30 minutes), as the rate of glycolysis is approximately 7 mg% per hour at room temperature.
For plasma separation following anticoagulants may be used.
Sodium Fluoride is preferred as anticoagulant due to its antiglycolytic activity. The higher concentration of Sodium fluoride i.e. more than 10 mg/ml blood should be avoided as it may inhibit the colour development. Glucose is stable for 24 hours in neatly separated plasma and serum. If the estimation is not possible within 24 hours then the specimen should be preserved at -10 degree C and should be used within 30 days.
PROCEDURE FOR END-POINT
1.0 ml procedure
Serum / Plasma
Incubate the assay mixture for 7 minutes at 37 degree C or 15 minutes at room temperature (25 - 30 degree C). After completion of incubation period measure the absorbance against blank at 505 nm. Final colour is stable for at least two hours if not exposed to direct light.
PROCEDURE FOR KINETIC ASSAY
Perform the assay as given below :
1.0 ml procedure
First carry out the assay of standard.
Mix and start stop watch simultaneously. Record absorbance at exactly 20th second after standard addition and then again at 60 degree second.
Subsequently, carry out the assay of the specimen following exactly the same procedure mentioned above.
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