Simple and sensitive liquid chromatographic quantitative analysis of the novel marine anticancer drug Yondelis™ (ET-743, trabectedin) in human plasma using column switching and tandem mass spectrometric detection
Simple and sensitive liquid chromatographic quantitative analysis of the novel marine anticancer drug Yondelis™ (ET-743, trabectedin) in human plasma using column switching and tandem mass spectrometric detection
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DOI：
10.1002/jms.608
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发表时间：
2004-04-01
影响因子：
2.3
通讯作者：
Beijnen, JH
中科院分区：
文献类型：
--
作者：
Stokvis, E;Rosing, H;Beijnen, JH
建立了一种简便、灵敏的人血浆中海洋抗癌药物Yondelis(ET-743，Trabectedin)的高效液相色谱-串联质谱仪(MS/MS)定量分析方法。蛋白质用甲醇沉淀后，稀释后的提取物进样到一个小的LC柱(10×3.0 mm i.d.)用于在线浓缩和进一步净化样品。接下来，分析物和氚内标物被反冲洗到分析柱上，以便在API2000三重四极杆质谱仪中进行分离和随后的检测。该方法的最低检测下限为0.05ngmL(-1)，使用100mul血浆，线性动态范围可达2.5ngmL1。方法的验证是根据FDA最新的生物分析方法验证指南进行的。与现有的测定人血浆中ET-743的LC/MS/MS方法相比，脱机样品制备所需的时间减少了10倍，采用了劳动密集型固相萃取程序进行样品前处理。所提出的柱切换方法已成功地应用于Yondelis和药动学监测的II期临床试验。版权所有(C)2004 John Wiley Sons，Ltd.
The development of a simple and sensitive assay for the quantitative analysis of the marine anticancer agent Yondelis (ET-743, trabectedin) in human plasma using liquid chromatography (LC) with column switching and tandem mass spectrometric (MS/MS) detection is described. After protein precipitation with methanol, diluted extracts were injected on to a small LC column (10 x 3.0 mm i.d.) for on-line concentration and further clean-up of the sample. Next, the analyte and deuterated internal standard were back-flushed on to an analytical column for separation and subsequent detection in an API 2000 triple-quadrupole mass spectrometer. The lower limit of quantitation was 0.05 ng mL(-1) using 100 mul of plasma with a linear dynamic range up to 2.5 ng ml(-1). Validation of the method was performed according to the most recent FDA guidelines for bioanalytical method validation. The time needed for off-line sample preparation has been reduced 10-fold compared with an existing LC/MS/MS method for ET-743 in human plasma, employing a labor-intensive solid-phase extraction procedure for sample pretreatment. The proposed column switching method was successfully applied in phase II clinical trials with Yondelis and pharmacokinetic monitoring. Copyright (C) 2004 John Wiley Sons, Ltd.