On-line extraction coupled with liquid chromatography tandem mass spectrometry for quantitation of pravastatin and its metabolite in human serum
On-line extraction coupled with liquid chromatography tandem mass spectrometry for quantitation of pravastatin and its metabolite in human serum
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DOI：
10.1002/bmc.989
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发表时间：
2008-07-01
影响因子：
1.8
通讯作者：
Wang, Yongcheng
中科院分区：
文献类型：
--
作者：
Wang, Xiaodong;Wang, Yongcheng
建立了一种高通量同时测定人血清中普伐他汀及其代谢物（M1）的生物分析方法，并采用在线萃取-液相色谱-串联质谱（LC-MS/MS）技术进行了验证。通过将50 μ L血清样品（与内标物水溶液以4：1混合）以3 mL/min的流速直接进样至其中一个含1 mm甲酸水溶液的萃取柱中，完成在线萃取。通过将分析物从萃取柱和分析柱反洗脱至质谱仪，以0.8 mL/min的流速使用由62% 1 mM甲酸水溶液和38%乙腈组成的等度移动的流动相进行分离和分析。在第一个柱用于分析时，平衡第二个萃取柱，反之亦然。普伐他汀和M1的标准曲线范围为0.500-100 ng/mL。当使用50 μ L人血清时，所有分析物的定量下限均为0.500 ng/mL。日内和日间精密度在7.4%以内，准确度在95%~ 103%之间。在线提取在0.5 min内完成，每个样品的总分析时间为2.5 min。版权所有（c）2008约翰威利父子有限公司。
A high-throughput bioanalytical method for simultaneous quantitation of pravastatin and its metabolite (M1) in human serum was developed and validated using on-line extraction following liquid chromatography tandem mass spectrometry (LC-MS/MS). The on-line extraction was accomplished by the direct injection of a 50 mu L serum sample, mixed 4:1 with an aqueous internal standard solution, into one of the extraction columns with aqueous 1 mm formic acid at flow rate of 3 mL/min. The separation and analysis were achieved by back-eluting the analytes from the extraction column and the analytical column to the mass spectrometer with an isocratic mobile phase consisting of 62% aqueous 1 mM formic acid and 38% acetonitrile at a flow rate of 0.8 mL/min. The second extraction column was being equilibrated while the first column was being used for analysis, and vice versa. The standard curve range was 0.500-100 ng/mL for pravastatin and M1. The lower limit of quantitation, 0.500 ng/mL for all the analytes, was achieved when 50 mu L of human serum was used. The intra- and inter-day precisions were within 7.4%, and the accuracy was between 95 and 103%. The on-line extraction was finished in 0.5 min and total analysis time was 2.5 min per sample. Copyright (c) 2008 John Wiley & Sons, Ltd.