Clinical Chemistry
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Clinical Chemistry 35: 2039-2043, 1989;
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Clinical Chemistry, Vol 35, 2039-2043, Copyright © 1989 by American Association for Clinical Chemistry

Simple enzymatic method for determining 1,5-anhydro-D-glucitol in plasma for diagnosis of diabetes mellitus

M Yabuuchi, M Masuda, K Katoh, T Nakamura and H Akanuma
Research Laboratory, Nippon Kayaku Co. Ltd., Tokyo, Japan.

We have developed a simple method for determination of 1,5-anhydro-D- glucitol in plasma, based on use of pyranose oxidase (EC 1.1.3.10), an enzyme with specificity toward pyranoid compounds such as 1,5-anhydro-D- glucitol and glucose. Plasma samples deproteinized with trichloroacetic acid are passed through a two-layer mini-column packed with strongly basic anion (OH- form, the upper layer) and strongly acidic cation (H+ form, the lower layer) exchange resins. 1,5-Anhydro-D-glucitol is efficiently recovered in the flow-through fraction, which is almost devoid of other sugars that are sensitive to pyranose oxidase. The hydrogen peroxide formed in the enzymatic oxidation of 1,5-anhydro-D- glucitol is detected by a standard method utilizing an enzymatic color- developing system. The overall assay system is highly specific for 1,5- anhydro-D-glucitol. The correlation between results obtained in the present method (x) and in the gas-liquid chromatographic (GLC) method (y) was: y = 1.062x-0.293 mg/L (r = 0.997, n = 49, Sxy = 10.78 mg/L). Compared with GLC, our method is simpler in the sample treatment step and quicker in the measuring step. The precisions of the two methods are comparable.


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 Appl. Environ. Microbiol.Home page
N. Yoshida, E. Uchida, T. Katsuragi, and Y. Tani
A Novel NAD-Dependent Dehydrogenase, Highly Specific for 1,5-Anhydro-D-Glucitol, from Trichoderma longibrachiatum Strain 11-3
Appl. Envir. Microbiol., May 1, 2003; 69(5): 2603 - 2607.
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Copyright © 1989 by the American Association for Clinical Chemistry.