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采用毛细管电泳-电动色谱法筛选中药中的α-葡萄糖苷酶抑制剂。

Screening α-glucosidase inhibitors from traditional Chinese drugs by capillary electrophoresis with electrophoretically mediated microanalysis.

机构信息

School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, PR China.

出版信息

J Pharm Biomed Anal. 2010 Dec 15;53(5):1250-3. doi: 10.1016/j.jpba.2010.07.041. Epub 2010 Aug 6.

Abstract

In the present study, we report the study by a combination of electrophoretically mediated microanalysis method with a partial technique for screening alpha-glucosidase inhibitors from 21 traditional Chinese drugs. In the setup, substrates and enzymes were introduced into the capillary as distinct plugs, the electrophoretic conditions for enzyme reaction and separation of substrates and products were different in the composition and pH of the background electrolyte, which make more enzyme reactions possible. Part of the capillary was filled with the optimal buffer for the enzyme reaction, whereas the rest was filled with the background electrolyte optimal for the separation of substrates and products. With the optimal condition, the Michaelis-Menten constant and the inhibitive mechanism of acarbose were studied, which were in the same range as previous literature data. Furthermore, the inhibitory ratios of enzymatic activity (IRE) of 21 traditional Chinese drugs were determined. The classical method has superiorities over traditional assay methods, which not only minimizes the false-positive results but also simplifies the experimental processes. It could be used for screening inhibitors in natural extract.

摘要

在本研究中,我们报告了一种组合电泳介导的微分析方法与一种筛选α-葡萄糖苷酶抑制剂的部分技术相结合,从 21 种中药中筛选α-葡萄糖苷酶抑制剂。在该装置中,底物和酶分别作为不同的电泳流进毛细管,酶反应和底物与产物分离的电泳条件在背景电解质的组成和 pH 值上有所不同,这使得更多的酶反应成为可能。毛细管的一部分填充有酶反应的最佳缓冲液,而其余部分填充有最佳分离底物和产物的背景电解质。在最佳条件下,研究了阿卡波糖的米氏常数和抑制机制,与以前的文献数据在同一范围内。此外,还测定了 21 种中药的酶活性抑制率(IRE)。与传统的检测方法相比,经典方法具有优势,不仅可以最大限度地减少假阳性结果,而且还可以简化实验过程。它可用于筛选天然提取物中的抑制剂。

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