Analytical Sciences

A Novel Method for the Assay of alpha-Glucosidase Inhibitory Activity Using a Multi-channel Oxygen Sensor

Kiyoshi MATSUMOTO,*  Kayoko TAKEMATA,* Kiyofumi TAKAYAMA,* Kanthi J. M. ABESUNDARA,* Toshiro MATSUI,* and Hideo KATAYAMA**

*Laboratory of Food Analysis, Division of Food Biotechnology, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan
**Daikin Industrial Co., Ltd., Miyukigaoka, Tsukuba 305-841, Japan

The quantification of glucose by using a multi-channel dissolved oxygen (DO) meter (DOX96) with immobilized glucose oxidase (GOD) and mutarotase (MUT) was performed. An evaluation of the inhibitory activities for alpha-glucosidase (AGH) by modifying our batch-type pseudo-in vivo assay system [Oki et al.; Biol. Pharm. Bull., 2000, 232, 1084] was also performed using a DOX96. When 45 U/well GOD and 18.75 U/well MUT were immobilized on the surface of a gelatin membrane on the electrodes, the response shown by the decrease percent of DO (%) obtained with 8 electrode wells in the same row was linear with the glucose concentration up to 3.3 mM and a correlation coefficient larger than 0.9. To estimate the AGH inhibitory activity, AGH-immobilized Sepharose supports in the well of a silent screen plate were used. The IC₅₀ values of acarbose and 1-deoxynojirimycin, a medicinal inhibitor for diabetes, were 0.70 ± 0.08 uM and 0.40 ± 0.13 uM, respectively, and coincided well with those by a pseudo-in vivo assay.

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