Fully enzymatic method for determining 1,5-anhydro-D-glucitol in serum.
We have developed a fully enzymatic method to measure 1,5 anhydro-D-glucitol (1,5-AG) in serum through use of pyranose oxidase (PROD: EC 1.1.3.10), glucokinase (EC 2.7.1.2), and an ATP-regenerating system. In a previous report (Clin Chem 1989;35:2039-43) the glucose interfering with the measurement of 1,5-AG was removed with a minicolumn. In the method used here, glucokinase and an ATP-regenerating system efficiently convert glucose to the unreactive compound, glucose 6-phosphate, making the method selective for 1,5-AG. The hydrogen peroxide produced in the oxidation of 1,5-AG by PROD is detected with a standard enzymatic color-developing system. The within-run and day-to-day precision (CV) of this method was 0.52-1.29% and 1.17-4.48%, respectively. The correlation (r) between the results obtained with our proposed method (y) and those obtained with the mini-column method (x) was 0.998 (y = 1.007x + 0.493 mg/L; n = 100; Sy/x = 0.641 mg/L). This newly developed method allows quicker and easier measurement of serum 1,5-AG than previously described methods.
Department of Laboratory Medicine, Gunma University School of Medicine, Japan.
Y
Y
Fukumura
S
S
Tajima
S
S
Oshitani
Y
Y
Ushijima
I
I
Kobayashi
F
F
Hara
S
S
Yamamoto
M
M
Yabuuchi
D000255:Adenosine Triphosphate / Q000378:metabolism; D001322:Autoanalysis / Q000379:methods* / Q000706:statistics & numerical data; D002237:Carbohydrate Dehydrogenases / Q000378:metabolism*; D003124:Colorimetry / Q000706:statistics & numerical data; D003847:Deoxyglucose / Q000097:blood*; D005941:Glucokinase / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis / Q000378:metabolism; D010728:Phosphoenolpyruvate / Q000378:metabolism; D011770:Pyruvate Kinase / Q000378:metabolism; D011786:Quality Control; D012680:Sensitivity and Specificity
C006584:1,5-anhydroglucitol; D010728:Phosphoenolpyruvate; D000255:Adenosine Triphosphate; D003847:Deoxyglucose; D006861:Hydrogen Peroxide; D002237:Carbohydrate Dehydrogenases; C022489:pyranose oxidase; D005941:Glucokinase; D011770:Pyruvate Kinase