液相色谱-串联质谱法对小鼠血浆中远志皂苷元的测定

Determination of Senegenin in Mouse Plasma by Liquid Chromatography-Tandem Mass Spectrometry

建立了小鼠血浆中远志皂苷元(senegenin,Se)的高效液相色谱-串联质谱(HPLC-MS/MS)测定方法。以RESTEK PinnacleⅡC18柱(100mm×2.1mm,5μm)为色谱柱,乙腈-水(体积比51∶49)为流动相,流速200μL.min-1;柱温20℃;质谱条件为气动辅助电喷雾离子源(ESI),检测方式为负离子多反应监测模式(MRM),用于定性分析的离子对为m/z535/481和m/z535/423,其中m/z535/481为定量离子;生物样品采用固相萃取方法处理。远志皂苷元标准曲线的线性范围为0.5～1000μg.L-1,定量下限达0.5μg.L-1,日内、日间相对标准偏差(RSD)均小于15%,回收率大于80%,精密度和准确度等均符合生物样品分析的要求。结果表明该法准确、灵敏、特异,适用于血浆中远志皂苷元的测定。该文还借助ABI4000 Q TRAP四极杆质谱仪特有的MS/MS/MS技术探讨了远志皂苷元碰撞活化裂解(collision-active dissociation,CAD)主要质谱碎片的产生机理。

A method for the determination of senegenin in mouse plasma was established by liquid chromatography-tandem mass spectrometry （HPLC - MS/MS）. RESTEK Pinnacle Ⅱ C18 column （100 mm ×2. 1 mm, 5μm） was used with a mobile phase of acetonitrile-water（51 ： 49, by volume）. The column temperature was 20 ℃. Gas auxiliary electrospray ionization source（ESI） was used to detect the senegenin by the negative electrospray ionization and multiple reaction monitoring mode（MRM） with m/z 535/481 and m/z 535/423 as the qualitative ions and m/z 535/481 as the quantitative ions. The plasma samples were extracted by solid phase extraction. The calibration curves of senegenin in plasma were linear in the range of 0.5 to 1 000 μg·L^-1, and the quantitation limit was 0.5 μg·L^-1. The intra-day and inter-day RSDs were less than 15% . The recoveries were more than 80% . Results showed that the method established was accurate, sensitive and specific, and suitable for the simultaneous determination of senegenin in mouse plasma. The collision-active dissociation（CAD） pathway of senegenin was also discussed.