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

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。

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

Objective To establish a simple and rapid LC-MS/MS method for determining the concentrations of paclitaxel and docetaxel in human plasma.Methods Roxithromycin was used as an internal standard,plasma samples were deproteinized by adding acetonitrile and centrifuged before analysis.The separation was achieved on a Hypersil GOLD aQ column with a mobile phase composed 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.Results Total run time of each plasma sample containing paclitaxel and docetaxel was 6 min.The proposed method was linear over the range 0.005-1.000 μg·mL-1 for both paclitaxel and docetaxel, and the lower limit of quantification (LLOQ) was 0.005 μg·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 ℃ for 2 h and at -80 ℃ for up to 1 month.Conclusion A simple,rapid,sensitive and accurate LC-MS/MS method for simultaneous determination of paclitaxel and docetaxel in human plasma was developed,and was successfully applied to therapeutic drug monitoring.