Print

DISCRETPAK™ REAGENTS - Glucose Hexokinase

Intended Use

For IN VITRO diagnostic use in the quantitative determination of γ-Glucose in serum using manual or automated applications.

Method Principle

The enzyme Hexokinase catalyzes the phosphorilation of D-glucose in the presence of ATP and Magnesium ions. The resultant glucoase-6-phosphate is then oxidized to 6-phosphoglucono lactone with concomitant reduction of Nicotinamide adenine dinucleotide (NAD). The amount of NADH produced is proportional to the glucose present in the serum sample and it is quantitated at 340 nm. The reaction scheme below illustrates the reactions that occur in this method.

HK
                            D-glucose + ATP ─────> D-gluc-6-phosphate + ADP + H

G6PDH
         D-glucose-6-phosphate + NAD ─────>6-phosphoglucono lactone + NADH


Method Performance Characteristics

Sensitivity: 0.0008 - 0.0012 absorbance units per mg/dL.
Linear Range: 0-600 mg/dL.
Precision: Within-run and day-to-day precision is summarized below.

Glucose

Within-Run Precision

Total Precision

MEAN

SD

CV

SD

CV

mg/dL

mg/dL

%

mg/dL

%

58

1.30

2.40

3.10

5.20

289

1.80

0.70

7.40

2.50

474

3.50

0.70

5.00

1.00


Correlation

A comparison of this method using an automated analyzer and a reference method based upon the hexokinase-G6PDH reaction resulted in the following regression statistics.

Correlation Data

Parameter

Data Observed

N

160

Range (mg/dL)

60-472

Regression

Y = 0.993x + 2.0

Correlation

  r = 0.999

Sy,x

2.9