分析化学2013，Vol.41Issue(7)：1013-1018,6.DOI:10.3724/SP.J.1096.2013.20961
超高效液相色谱串联质谱联用法快速筛选3-羟基-3-甲基戊二酰辅酶A还原酶抑制剂
Rapid Screening of 3-Hydroxy-3-methylglutaryl CoA Reductase Inhibitors by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry
摘要
Abstract
A rapid and accurate method has been developed for screening 3-hydroxy-3-methylglutaryl CoA reductase (HMGCR) inhibitors by measuring the concentration of reduced nicotinamide adenine dinucleotide phosphate (NADPH) in the enzymatic reaction system using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).Several parameters,including the enzymatic reaction system,liquid chromatography and mass spectrometric conditions were optimized for improving the chromatographic performance and MS signal.The effect of ionization reagent and the fragmentation mechanism of NADPH were also discussed.The separation was performed on an ACQUITY UPLC BEH Amide column (100 mm × 2.1 mm,1.7 μm) with a mobile phase consisting of triethylamine/hexafluoroisopropanol buffer at a flow of 0.3 mL/min,column temperature of 30 ℃.Identification and quantification were achieved by UPLC-MS/MS in an electrospray ionization-negative mode and multiple reactions monitoring (MRM).A good linearity was obtained for determination of NADPH at the concentration of 0.05-50 μmol/L with the linear correlation coefficient more than 0.9996.The limit of quantification for NADPH was 0.05 μmol/L (S/N=10).关键词
超高效液相色谱串联质谱/还原型烟酰胺腺嘌呤二核苷酸磷酸/3-羟基-3-甲基戊二酰辅酶A还原酶/抑制剂筛选Key words
Ultra performance liquid chromatography-tandem mass spectrometry/ Reduced nicotinamide adenine dinucleotide phosphate/ 3-Hydroxy-3-methylglutaryl CoA reductase/ Inhibitor screening引用本文复制引用
吴琼,王世聪,马俊锋,李婉南,邢述,付学奇..超高效液相色谱串联质谱联用法快速筛选3-羟基-3-甲基戊二酰辅酶A还原酶抑制剂[J].分析化学,2013,41(7):1013-1018,6.基金项目
本文系教育部博士点基金项目(No.20090061110019)资助 （No.20090061110019）
