Determination of 1,5-anhydroglucitol in urine by high performance liquid chromatography and an enzyme sensor.
A simple high performance liquid chromatographic method combined with an enzyme sensor has been developed to measure 1,5-anhydroglucitol in urine. The enzyme sensor consists of a hydrogen peroxide electrode and a chitosan membrane of an immobilized pyranose oxidase. As the system does not resist interfering substances, urine samples are first purified by passing them through a two-layer column packed with (1) strongly basic anion (OH- form, the upper layer) and (2) strongly acidic cation (H+ form, the lower layer) exchange resins. 1,5-Anhydroglucitol is efficiently recovered in the flow-through fraction of the column. In this system, the minimum detectable concentration of 1,5-anhydroglucitol is 0.1 mg/L, and the measurable range extends from 0.1 to 60 mg/L. The coefficient of variation values of the within-day and day-to-day precisions are 3.0-6.5% and and 4.4-6.7% respectively, and there is good agreement between the results measured by our method and those obtained by the gas-liquid chromatographic/mass spectrometric method (r = 0.994). The method we have described here has been successfully used to elucidate a mechanism for the reducing 1,5-anhydroglucitol level in the serum and plasma of patients.
Tajima
S
S
Takasaki Research Laboratories, Nippon Kayaku Co. Ltd., Gumma, Japan.
Hashiba
M
M
Suzuki
T
T
Akanuma
H
H
Yabuuchi
M
M
D015374:Biosensing Techniques; D002237:Carbohydrate Dehydrogenases / Q000737:chemistry; D002648:Child; D002686:Chitin / Q000031:analogs & derivatives; D048271:Chitosan; D002851:Chromatography, High Pressure Liquid; D003847:Deoxyglucose / Q000097:blood / Q000652:urine*; D004800:Enzymes, Immobilized; D008401:Gas Chromatography-Mass Spectrometry; D005947:Glucose; D006801:Humans; D006861:Hydrogen Peroxide; D007202:Indicators and Reagents; D008297:Male
D004800:Enzymes, Immobilized; D007202:Indicators and Reagents; D002686:Chitin; C006584:1,5-anhydroglucitol; D048271:Chitosan; D003847:Deoxyglucose; D006861:Hydrogen Peroxide; D002237:Carbohydrate Dehydrogenases; C022489:pyranose oxidase; D005947:Glucose