LC-MS/MS法同时测定人血浆中紫杉醇和多西他赛的质量浓度

Simultaneous determination of paclitaxel and docetaxel in human plasma by LCMS/MS

目的建立一种简单、快速,可同时测定人血浆中紫杉醇和多西他赛药物质量浓度的LC-MS/MS方法。方法以罗红霉素为内标,血浆经乙腈沉淀蛋白、离心后进样分析;色谱柱为Hypersil GOLD aQ;流动相为1mL·L-1甲酸溶液-乙腈;梯度洗脱;流速为0.4mL·min-1;离子源为可加热电喷雾离子源(HESI);检测方式:正离子检测;扫描方式为多反应监测(MRM),紫杉醇、多西他赛和罗红霉素的检测离子对分别为m/z854.3→509.1,808.3→527.1和837.6→679.3。结果每份含紫杉醇和多西他赛的样品分析时间为6min,血浆中紫杉醇和多西他赛的药物质量浓度在0.005~1.000μg·mL-1范围内线性关系良好,最低定量限为0.005μg·mL-1,两药测定的日内和日间精密度RSD<15%,提取回收率分别为94.97%~101.95%和96.07%~112.86%,基质效应分别为101.14%~115.59%和82.40%~95.77%。紫杉醇和多西他赛血浆样品在反复冻融3次、4℃保存2h和-80℃保存1个月的条件下稳定。结论该研究建立的LC-MS/MS分析方法简便、快速、灵敏和准确,能同时测定人血浆中紫杉醇和多西他赛的质量浓度。

To establish a simple and rapid LC-MS/MS method for determining the concentrations of paclitaxel and do-cetaxel in human plasma. Roxithromycin was used as an internal standard, plasma samples were deproteinized by addingacetonitrile and centrifuged before analysis. The separation was achieved on a Hypersil GOLD aQ column with a mobile phase com-posed of 1 mL ？ L 1 formic acid and acetonitrile in gradient elution mode at a flow rate of 0. 4 mL · min 1. The ionization source was the heated electrospray ionization source （HESI） ？ and multiple reaction monitoring （MRM） in positive ion mode was used to detect the analytes. The precursor-product ion pairs used for MRM were m /z 854. 3-509. 1 , 808. 3- 527. 1 and 837. 6- 679. 3 for paclitaxel,docetaxel and internal standard roxithromycin,respectively. Total run time of each plasma sample containing pa-clitaxel and docetaxel was 6 min. The proposed method was linear over the range 0. 005-1. 000 jug · mL 1 for both paclitaxel and docetaxel, and the lower limit of quantification （LLOQ） was 0. 005 jng＊mL 1 for the analytes. The intraday and interday precision was within 15%,the recovery ranged from 94. 97% to 101. 95% for paclitaxel and from 96. 07% to 112. 86% for docetaxel, and the matrix effects of paclitaxel and docetaxel were within 101. 14%-115. 59% and 82. 40%-95. 77% , respectively. Paclitaxel and docetaxel were stable after freeze-thaw cycles 3 times and during storage at 4 °C for 2 h and at -80 °C for up to 1 month.A simple, rapid, sensitive and accurate LC-MS/MS method for simultaneous determination of paclitaxel and docetaxel in hu-man plasma was developed, and was successfully applied to therapeutic drug monitoring.