LC-MS/MS定量分析小鼠血浆和组织液中多西他赛的含量

LC-MS/MS for Quantitative Analysis of Docetaxel in Mice Plasma and Tissue Fluid

目的:建立小鼠血浆和组织样品中多西他赛的含量测定方法。方法:选择紫杉醇为内标物,进样前用甲基叔丁基醚萃取,Hypersil BDS C_(18)色谱柱(2.1 mm×50 mm,2.4μm),流动相甲醇-0.1%甲酸(80∶20),柱温40℃,流速0.3 m L·min^(-1),进样量10μL。电喷雾离子源,多反应检测模式进行正离子检测,离子源喷雾电压3.5 kV,加热毛细管温度320℃,鞘气压力35 Pa,辅助气压力10 Pa,碰撞能量30 V,碰撞诱导解离电压12 V,多西他赛检测离子对m/z 876.3~549.1,紫杉醇检测离子对m/z 830.3~307.9。结果:多西他赛在小鼠血浆中的线性范围0.013~13 mg·L^(-1),在心、肝、脾、肺、肾的线性范围分别为0.012~12.937,0.013~13.068,0.012~13.001,0.013~13.133,0.011~12.965 mg·L^(-1);日内及日间精密度、各条件下稳定性均<15%;血浆中低、中、高质量浓度多西他赛的平均回收率分别为98.24%,97.51%,93.70%;在小鼠心、肝、脾、肺、肾各组织的平均回收率在87.87%~98.90%。结论:相较于HPLC,LC-MS/MS更适合检测小鼠血浆及组织液中多西他赛的含量,该方法稳定可行、重复性好。

Objective： To establish a determination for docetaxel that derives from mice plasma and tissue fluid samples,and evaluate effectiveness for this method. Method： Samples were purified by methyl tertbutyl ether with paclitaxel as internal standard. Chromatographic separation was performed on a Hypersil BDS C18 column using a mobile phase composed of methanol-0. 1% formic acid（ 80∶ 20）,column temperature was 40 ℃,flow rate was 0. 3 m L ·min-1. Detection was performed on atriple-quadrupole tandem mass spectrometer with multiple reaction monitoring（ MRM） mode via electrospray ionization（ ESI） source,ESI voltage was 3. 5 k V,capillary temperature was 320 ℃,sheath gas pressure was 35 Pa,auxiliary gas pressure was 10 Pa,collision energy was 30 V,CID pressure was 12 V,detection ion for docetaxel was m / z 876. 3-549. 1,detection ion for paclitaxel was m / z 830. 3-307. 9. Result： Docetaxel in mice plasma, heart, liver, spleen, lung and kidney showed good linear relationship within 0. 013-13, 0. 012-12. 937, 0. 013-13. 068, 0. 012-13. 001, 0. 013-13. 133,0. 011-12. 965 mg·L-1. RSD of intra-day and inter-day assays were less than 15%. Recovery rates of docetaxel with low,middle and high concentration in plasma were 98. 24%,97. 51% and 93. 70%,its recovery rates in heart,liver,spleen,lung and kidney were 87. 87%-98. 90%. Conclusion： Compared with HPLC,LCMS / MS is more suitable for quantitative analysis of docetaxel in mice plasma and tissue fluid samples.