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 con...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.展开更多
基金supported by the National Basic Research Program of China(973 Program)(No.2012CB724003)
文摘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.
