HPLC-MS/MS法测定大鼠血浆中紫杉醇的含量Determine Paclitaxel in Rat Plasma by HPLC-MS/MS Method
石永辉,余晓霞,吕立,李国成,邱凯锋
SHI Yonghui,YU Xiaoxia,LV Li,LI Guocheng,QIU Kaifeng
摘要(Abstract)：
目的建立测定大鼠血浆中紫杉醇浓度的高效液相色谱-串联质谱(HPLC-MS/MS)法,为研究紫杉醇的药物动力学提供方法学基础。方法以多西他赛为内标,色谱柱为Agilent Eclipse-plus C_(18)柱(4.6 mm×50 mm,3.8μm),流动相为甲醇-水(含0.1%的甲酸,80∶20,V/V),流速为0.5 m L·min~(-1),采用电喷雾离子源,扫描方式为多重反应监测(MRM),正离子模式检测。结果紫杉醇浓度在2~1 000 ng·m L~(-1)范围内线性关系良好(R~2>0.99),定量下限为2 ng·m L~(-1),其中低、中、高3种浓度(6,300,750 ng·m L~(-1))的平均回收率为(109.10±6.97)%,(97.51±0.57)%及(97.98±2.95)%,日内及日间精密度的相对标准偏差(RSD)<15%。结论本方法操作灵敏、快捷、结果准确,可用于测定大鼠体内紫杉醇的浓度。
OBJECTIVE To establish and validate a method for quantification of paclitaxel in rat plasma by a tandem mass spectrometry detection( HPLC-MS/MS) and to provide method in studying the pharmacokinetics of paclitaxel in rats. METHODS Paclitaxel for rats plasma and docetaxel as internal standard were extracted with methanol and separated on an Agilent Eclipse-plus C_(18)( 4.6 mm × 50 mm,3. 8 μm) using mobile phase of methanol-water( include 0. 1% formic acid,80 ∶ 20,V/V) at a flow rate of 0.5 m L·min~(-1). Mass spectrometry detection was operated in positive ion mode with multiple reaction monitoring( MRM) performing.RESULTS The linear range of paclitaxel in plasma was 2-1 000 ng·m L~(-1) with good correlation( R~2> 0. 99). The lower limit of quantification was 2 ng·m L~(-1)( S/N>10). The average recoveries of paclitaxel( 6,300,750 ng·m L~(-1)) were 109.10%±6.97%,97.51%±0.57% and 97.98% ± 2. 95% respectively. Both intra-and inter-day precisions( as relative standard deviation,RSD) were < 15%.CONCLUSION The method is sensitive,rapidly and accurate. It can be used to determine the concentration of paclitaxel in rats.
关键词(KeyWords)：
紫杉醇;高效液相色谱-串联质谱法
paclitaxel;HPLC-MS/MS
基金项目(Foundation): 广东省自然科学基金项目(2014A030310394、2016A030313330);; 国家自然科学基金(81503001);; 广东省省级科技计划项目(2016A020215069、2016A020215063);; 广东省中医药局科研项目(20161061);; 广东省医院药学研究基金(2015ZX001)
作者(Author):
石永辉,余晓霞,吕立,李国成,邱凯锋
SHI Yonghui,YU Xiaoxia,LV Li,LI Guocheng,QIU Kaifeng
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