HPLC-DAD法同时测定延胡索中4种生物碱的含量

Determination of four alkaloids in corydalis yanhusuo by HPLC-DAD

目的:采用高效液相色谱-二极管阵列检测器(high performance liquid chromatography-diode array detection,HPLC-DAD),建立同时测定延胡索药材中4种生物碱含量的方法,并比较不同产地中4种生物碱含量的差异。方法:采用Agilent Eclipse XDBC18(5μm,4.6 mm×250 mm)色谱柱,以乙腈-0.05%磷酸(pH 6.0)为流动相进行梯度洗脱,流速1.0 ml/min,检测波长分别为巴马亭、紫堇碱270 nm,延胡索乙素和四氢小檗碱280 nm。结果:巴马亭在1~10μg/ml、紫堇碱在10~100μg/ml、延胡索乙素在5~30μg/ml,四氢小檗碱在1~10μg/ml浓度范围内线性关系良好,高、中、低3种浓度平均加样回收率在98.2%~101.3%之间。结论:本方法操作简单、稳定、重复性好,可用于同时测定延胡索中4种生物碱的含量。

Objective：To establish a high performance liquid chromatography-diode array detection （HPLC-DAD） method for the si- multaneous determination of four alkaloids in corydalis yanhusuo, and to compare the contents of four alkaloids at different production places. Methods：Angilent Eclipse XDB-C18（5 μm,4.6 mm × 250 mm） was used as the analytical column. Acetonitrile-0.05% H^PO4（pH 6.0）was used as the mobile phase with gradient elution and the flow rate was 1.0 ml/min. The detection wavelengths were 270 nm for palmatine and corydaline, 280 nm for tetrahydropalmatine and tetrahydroberineper respectively. Results ： Palmatine, coryda- line, tetrahydropalmatine and tetrahydroberineper have a good linear relation within ranges of 1-10 μg/ml, 10-100 μg/ml, 5-30μg/ml, 1-10μg/ml,respectively. Spiked recovery of high, middle and low concentration ranged from 98.2% to 101.3%. Conclusion：The method is simple, accurate and reproducible and can be used to determine the above four alkaloids.