HPLC法同时测定延胡索中5种生物碱含量的建立及应用

Simultaneous determination of five alkaloids by HPLC and application analysis on Corydalis pieces and granules

目的建立HPLC法同时测定延胡索中原阿片碱、延胡索乙素、延胡索甲素、小檗碱、巴马汀等5种成分含量,并应用于延胡索饮片和颗粒剂的含量测定和比较。方法采用Inertsil ODS-3柱(250 mm×4.6 mm,5μm);流动相:甲醇-0.1%磷酸水溶液,梯度洗脱;流速1.0 m L·min^(-1);检测波长280 nm;柱温35℃;进样体积20μL。结果在上述色谱条件下,5种生物碱成分在55 min内完全分离,每种成分在各自的线性范围内线性关系良好(r>0.999);5种生物碱的平均回收率(n=6)分别为99.92%、101.82%、101.42%、98.26%和101.52%,RSD分别为1.71%、1.28%、1.61%、1.90%和0.94%。建立的方法成功用于24批延胡索饮片和市售颗粒剂的含量测定。结论该方法操作简便、快速、准确、重复性好,可用于延胡索饮片和颗粒剂的质量评价。

AIM To establish a simultaneous method for the determination of protopine, tetrahydropalma- tine, corydaline, berberine and palmatine in Corydalis yanhusuo by HPLC, for the content determination and com- parison of Corydalis pieces and granules. METHODS An Inertsil ODS-3 column （ 250 mm × 4.6mm, 5 μm） was used with the mobile phase containing methanol -0.1% phosphoric acid for gradient elution at the flow rate of 1.0 mL·min^-1, the detection wavelength was 280 nm, the column temperature was set at 35℃, and the injection volume was 20 μL. RESULTS Methodological observations indicated each ingredient with a good linear relation- ship in their respective linear range （ r 〉 0. 999） in this method, the average recoveries of five alkaloids （ n = 6） were 99.92% , 101.82% , 101.42% , 98.26% and 101.52% respectively, and the RSD was 1.71%, 1.28% , 1.61% , 1.90% and 0.94% respectively. The established method had been successfully applied for the determination of 24 batches of Corydalis pieces and granules. CONCLUSION The method is simple, rapid, accurate and reproducible, and can be used for quality control of Corydalis pieces and granules.