高效液相色谱-质谱法同时检测兔血浆中3种草乌成分

Simultaneous determination of three components of wild aconite （草乌） in rabbit plasma by high performance liquid chromatography-mass spectrometer

目的建立高效液相色谱-质谱检测血浆新乌头碱、乌头碱和次乌头碱浓度的方法。方法采用Agilent 1200系列的高效液相色谱仪和Bruker esquire HCT质谱仪检测。兔血浆在碱性条件下经乙醚萃取，以乙腈：0.1％碳酸氢铵（60：40）为流动相，ZORBAX SB-C18色谱柱分离，质谱电喷雾离子化源（ESI），选择离子监测、内标法定量。结果新乌头碱、乌头碱、次乌头碱3种物质的线性范围宽，线性关系良好，定量下限分别为1.0、0.4和0.5μg／L；血浆的萃取回收率高，在84.17％-98.06％；3种成分的日内、日间精密度相对标准偏差均小于10％。结论该色-质联用分析方法准确、灵敏度高，可用于急性草乌中毒患者的血浓度监控及药代动力学和毒代动力学研究。

Objective To establish a high performance liquid chromatography-mass spectrometer （HPLC-MS） method for simultaneous determination of mesaconitine, aconitine and hypaconitine in rabbit plasma. Methods The HPLC instrument was Agilent 1200 series and the MS was Bruker esquire HCT series. The analytical column was packed with ZORBAX SB-C18. The mobile phase was acetonitrile ：0.1% ammonium bicarbonate （60 ： 40） and the flow rate was 0.6 ml/min. The rabbit plasma was extracted by diethyl ether. The selected ion was determined by electrospray ionization （ESI^＋）. The peak area with the internal standard method was used for determination. Results Mesaconitine, aconitine and hypaconitine showed excellent linear relationship with wide linear range, and the limited determination were 1.0, 0.4, 0.5μg/L respectively. The plasma extraction recovery rate was high （84.17%—98.06%）, the intra-day relative standard deviation （RSD） and inter-day RSD were lower than 10% respectively at three concentrations for the three substances showing excellent stability. Conclusion The method is rapid, sensitive and can be used to monitor the blood concentration of toxic material in patients with Radix aconiti kusnezoffii（草乌） intoxication and to investigate pharmacokinetics and toxicokinetics.