Optimization,validation and application of an assay for the activity of HMG-CoA reductase in vitro by LC–MS/MS

作     者：Jing Wang Ji-Ye Sun Chun-Jie Sha Yu-Feng Shao Yan-Hong Liu You-Xin Li Zhen-Wen Duan Wan-Hui Liu 

作者机构：School of PharmacyYantai University State Key Laboratory of Long-acting and Targeting Drug Delivery System Shandong Luye Pharmaceutical Co. Ltd. Peking University WBL Biotech Co. Ltd. 

出 版 物：《Journal of Pharmaceutical Analysis》 (药物分析学报（英文版）)

年 卷 期：2015年第5卷第6期

页      面：383-388页

学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 1002[医学-临床医学] 0817[工学-化学工程与技术] 0703[理学-化学] 0702[理学-物理学] 10[医学] 

基　　金：supported by the National Basic Research Program of China(973 Program)(No.2012CB724003) 

主　　题：XuezhikangLC MS/MSHMG CoA reductaseMevalonolactoneQuality control 

摘      要：A stable HMG-CoA reductase （HMGR） reaction in vitro was developed by a sensitive, selective and precise liquid chromatography-tandem mass spectrometry （LC-MS/MS） method. The optimized enzyme reaction condition contained 1.5 lag of HMGR, 20 nM of NADPH with 50 rain of reaction time. The method was validated by several intraand inter-day assays. The production transitions of m/z 147.0/59.1 and m/z 154.0/59.1 were used to detect and quantify mevalonolactone （MVAL） and MVAL-DT, respectively. The accuracy and precision of the method were evaluated over the concentration range of 0.005- 1.000 lag/mL for MVAL and 0.010-0.500 lag/mL for lovastatin acid in three validation batch runs. The lower limit of quantitation was found to be 0.005 lag/mL for MVAL and 0.010 lag/mL for lovastatin acid. Intra-day and inter-day precision ranged from 0.95% to 2.39% and 2.26% to 3.38% for MVAL, 1.46% to 3.89% and 0.57% to 5.10% for lovastatin acid, respectively. The results showed that the active ingredients in Xuezhikang capsules were 12.2 and 14.5 mg/g, respectively. This assay method could be successfully anDlied to the oualitv control study of Xuezhikanu caosule for the first time.