HPLC同时测定生物样品中新乌头碱、乌头碱、次乌头碱的含量

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

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。

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, 5μm）. 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.