A Method for Determination of Hexokinase Activity by RP-HPLC

作     者：GUAN Yuxia WANG Jing SUN Jiaxian 

作者机构：College of Biological Science and Technology FuzhouUniversity Fuzhou 350002 Fujian China 

出 版 物：《Wuhan University Journal of Natural Sciences》 (武汉大学学报（自然科学英文版）)

年 卷 期：2011年第16卷第6期

页      面：535-540页

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 

基　　金：Supported by the Key Project of Science and Technology Department of Fujian Province (2008Y0052) 

主　　题：reverse-phase high-performance liquid chromatog-raphy(RP-HPLC) hexokinase enzyme activity assay 

摘      要：A method was established to determine the activity of hexokinase by reverse-phase high-performance liquid chromatography （RP-HPLC）. The activity of hexokinase was determined by detecting the amount of ADP in the reaction that converts glucose into glucose-6-phosphate. The separation degree of ATP and ADP was as good as 3.46 （separation degree^l.5）. In the range of 0.01-0.40 mmol/L, there was a good linear relationship between concentration of ADP and its peak areas （the correlation coefficient was 0.999）. The relative standard deviation（RSD） of intraday was 1.43%-1.46%, that of interday was 2.12%-2.15%. At 30℃ （optimal temperature） and pH7.0 （optimal pH）, the recovery rate of hexokinase was 95.3%-97.6%. The results showed that the method had good precision and high recovery rate, and the enzyme activity was determined through only one-step reaction. To some extent, this method can reduce the cost.