HPLC同时测定生物样品中新乌头碱、乌头碱、次乌头碱的含量

 目的建立同时测定血液、肝组织中新乌头碱(mesaconitine)、乌头碱(aconitine)、次乌头碱(hypaconitine)含量的HPLC分析方法。方法采用Welchrom C₁₈柱(4.6mm×250mm,5μm),以乙腈-0.03%四丁基氢氧化铵(用冰醋酸调至pH9.74)(58∶42)为流动相,流速为1.0mL·min^-1,检测波长为230nm。结果生物样品中新乌头碱在0.077～23mg·L^-1内线性关系良好(r=0.9998),乌头碱在0.083～25mg·L^-1内线性关系良好(r=0.9998),次乌头碱在0.08～24mg·L^-1内线性关系良好(r=0.9996)。生物样品中新乌头碱、乌头碱、次乌头碱的回收率不低于83.6%。生物样品中新乌头碱、乌头碱、次乌头碱的日内精密度和日间精密度分别在5.1%和9.3%以内。结论本方法简便、灵敏、准确,适用于血液、肝组织中新乌头碱、乌头碱、次乌头碱的分析。

Simultaneous Determination of Mesaconitine,Aconitine and Hypaconitine in Blood and Liver Speciman by HPLC

OBJECTIVE To establish a HPLC method for simultaneous determination of Mesaconitine,Aconitine and Hypaconitine in blood and liver speciman. METHODS The samples were separated on the column of Welchrom ODS (4.6 mm×250 mm, 5 μm). The mobile phase consisted of acetonitrile-0.03% tetrabutyl ammonium hydroxide (with glacial acetic acid to pH 9.74) (58∶42) at a flow rate of 1.0 mL·min^-1.The detection wavelength was at 230 nm. RESULTS Mesaconitine in biological samples were linear in the ranged of 0.077-23 mg·L^-1(r=0.999 8), and Aconitine was linear in the ranged of 0.083-25 mg·L-1 (r=0.999 8), and Hypaconitine was linear in the range of 0.08-24 mg·L^-1(r=0.999 6). The mean extraction recovery was over 83.6% in biological samples. The intra and inter-day precision of assay for Mspeciman,Aconitine and Hypaconitine were less than 5.1% and 9.3% in biological samples. CONCLUSION The method is simple, sensitive, accurate, and suitable for the determination of Mesaconitine,Aconitine and Hypaconitine in blood and liver speciman for toxicological and clinical pharmaceutical analysis.