高效液相色谱法测定4种乌头中药中乌头碱、新乌头碱和次乌头碱

Determination of aconitine,mesaconitine and hypaconitine in four kinds of Chinese traditional medicines of Aconitum by high performance liquid chromatography

目的建立能够同时测定4种乌头属中药(生川乌、制川乌、草乌和制草乌)中乌头碱、新乌头碱和次乌头碱3种有效成分的高效液相色谱法。方法采用高效液相色谱法测定4种乌头属中药中乌头碱、新乌头碱和次乌头碱的含量。流动相采用10 mmol/L乙酸铵水溶液∶乙腈(72∶28);检测波长:232 nm;色谱柱:ZORBAX Eclipse Plus C18柱;柱温:35℃;流速:1.5 ml/min。结果乌头碱、新乌头碱和次乌头碱在2.5-250μg/ml浓度范围内,均呈良好的线性关系,乌头碱线性方程:Y=4.9344X+0.7928(r=0.9997),新乌头碱线性方程:Y=7.6304X+4.8092(r=0.9997),次乌头碱线性方程:Y=7.1914X+4.5630(r=0.9997)。乌头碱、新乌头碱和次乌头碱精密度、稳定性和重复性相对标准偏差均满足测定要求。结论本研究建立的高效液相色谱法能够同时测定生川乌、制川乌、草乌和制草乌4种乌头属中药中乌头碱、新乌头碱和次乌头碱3种有效成分,且方法操作简便、快速。

Objective To develop a high performance liquid chromatography(HPLC)method for the detemination of aconitine,mesaconitine and hypaconitine from four kinds of Chinese traditional medicines of Aconitum(Radix Aconiti,Radix Aconiti Preparata,Radix Aconiti Kusnezoffii and Radix Aconiti Kusnezoffii Preparata).Methods The contents of aconitine,mesaconitine and hypaconitine in four kinds of Aconitum herbs were determined by HPLC.The mobile phase was 10 mmol/L ammonium acetate∶acetonitrile(72∶28),the detection wavelength was 232 nm,the chromatographic column was ZORBAX Eclipse Plus C18,the column temperature was 35℃,and the flow rate was 1.5 ml/min.Results The linear regression equations of aconitine,mesaconitine and hypaconitine showed good linear relationship in the concentration range of 2.5~250μg/ml,and the regression equation was Y=4.9344 X+0.7928 for aconitine(r=0.9997),Y=7.6304 X+4.8092 for neaconitine(r=0.9997),and Y=7.1914 X+4.5630 for hypaconitine(r=0.9997).The relative standard deviations(RSDs)of precision,stability and repeatability of aconitine,mesaconitine and hypaconitine met the determination requirements.Conclusion The HPLC method established in this study could simultaneously determine three effective components in four kinds of Aconitum,and it is simple and rapid.