Quantitative determination of pravastatin and R-416, its main metabolite in human plasma, by liquid chromatography-tandem mass spectrometry
Quantitative determination of pravastatin and R-416, its main metabolite in human plasma, by liquid chromatography-tandem mass spectrometry
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DOI：
10.1016/j.jchromb.2004.11.014
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发表时间：
2005-02-25
影响因子：
3
通讯作者：
Urasaki, Y
中科院分区：
文献类型：
--
作者：
Kawabata, K;Samata, N;Urasaki, Y
建立了一种快速、灵敏地测定人血浆中普伐他汀及其主要代谢产物R-416的定量方法。使用Bond Elut（R）C-8通过固相萃取法从血浆样品中提取分析物。该方法涉及使用液相色谱结合大气压化学电离（APCI）和选择反应监测（SRM）质谱。普伐他汀类似物R-122798用作内标（I.S.）。普伐他汀、R-416和I.S.的分离使用反相柱（C-18）完成。洗脱的组分通过APCI源（负离子）电离，随后通过高选择性三重四极杆质谱仪在SRM模式下检测。在0.1 ng/mL（定量下限，LLOQ）至100 ng/mL范围内获得线性标准曲线。普伐他汀和R-416在LLOQ下样品的试验内精密度（变异系数）分别为1.8%和1.6%。普伐他汀和R-416的试验内准确度值分别为95.8-107.6%和92.6-109.0%。在0.5、10和80 ng/mL浓度下测定所有分析物的质控（QC）样品的精密度和准确度。根据QC样品计算的试验内和试验间精密度在普伐他汀和R-416的10%和11%范围内。普伐他汀和R-416的总回收率分别为75.7-82.1%和68.6- 74.3%。人血浆中普伐他汀和R-416在-20 ℃冰箱中可稳定储存3周，在室温下可稳定储存6小时，在6 ℃下可稳定储存48小时。该测定方法已成功用于评价健康志愿者口服美伐他汀（R）后的普伐他汀和R-416水平。(C)2004 Elsevier B. V.保留所有权利。
A quantitative method was developed and validated for rapid and sensitive analysis of pravastatin and R-416, the main metabolite of pravastatin, in human plasma. The analytes were extracted from plasma samples by a solid phase extraction method using a Bond Elut(R) C-8. The method involved the use of liquid chromatography coupled with atmospheric pressure chemical ionization (APCI) and selected reaction monitoring (SRM) mass spectrometry. A pravastatin analog, R-122798, was used as the internal standard (I.S.). Separation of pravastatin, R-416 and the I.S. was accomplished using a reverse-phase column (C-18). The components eluted were ionized by the APCI source (negative ion) and subsequently detected by a highly selective triple quadrupole mass spectrometer in the SRM mode. Linear standard curves were obtained from 0.1 ng/mL (lower limit of quantification, LLOQ) to 100 ng/mL. The intra-assay precisions (coefficient of variation) for the samples at the LLOQ were 1.8% for pravastatin and 1.6% for R-416. The intra-assay accuracy values were 95.8-107.6% for pravastatin, and 92.6-109.0% for R-416, respectively. Precision and accuracy of quality control (QC) samples were determined at concentrations of 0.5, 10 and 80 ng/mL for all analytes. The intra- and inter-assay precision calculated from QC samples were within 10% for pravastatin and within 11% for R-416. The overall recoveries for pravastatin and R-416 were 75.7-82.1% and 68.6-74.3%, respectively. Pravastatin and R-416 were stable in human plasma for 3 weeks at -20degreesC in a freezer, up to 6 h at room temperature, and up to 48 h at 6degreesC. This assay method was successfully used to evaluate the pravastatin and R-416 levels in healthy volunteers following oral administration of Mevalotin(R). (C) 2004 Elsevier B.V. All rights reserved.