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

摘要:

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 XDB-C18（5 μm，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 1～10 μg/ml、紫堇碱在10～100 μg/ml、延胡索乙素在 5～30 μg/ml，四氢小檗碱在 1～10 μg/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。

Abstract:

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% H3PO4（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，5-30 μg/ml，1-10 μg/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．