HPLC法测定甲炎康泰颗粒柴胡皂苷b1、b2、迷迭香酸的含量

Determination of Saikosaponin b1, b2 and Rosmarinic Acid in Jiayan KangtaiGranules by HPLC

目的:建立并验证测定柴胡皂苷b1、b2、迷迭香酸含量的方法。方法:采用AgilentZORBAXSB—C18(4.6mm×250mm,5μm)色谱柱,柴胡皂苷流动相为甲醇-水(72:28),检测波长为254nm,迷迭香酸流动相为甲醇(B)-0.1%三氟乙酸水(A)(0~5min,35%B,5.01~15min,35%~40%B,15.01~30min,40%B),检测波长为330nm,体积流量1mL·min^-1,柱温35℃。结果:柴胡皂苷b1、b2、迷迭香酸分别在0.37~9.25μg(r=0.9999)、O.52~13μg(r=0.9999)、0.2—5.61μg(r=0.9999)范围内进样量与峰面积呈良好的线性关系;准确度实验平均回收率(n=6)分别为96.89%、99.01%、97.09%,RSD(%)分别为1.93%、1.51%、1.61%;重复性实验含量RSD(%)分别为1.88%、2.28%、1.65%。结论:所建的测定方法准确、简便、具有良好的重复性,可用于甲炎康泰颗粒的质量控制。

Obiective: To establish and validate a method for determining contents of saikosaponinb1, b2（SSb1,b2） and rosmarinic acid（RosA）. Method: HPLC method was employed on a Agilent ZORBAX SB-C18 column（4.6 mm×250 mm, 5 μm）. The mobile phase for SSb was methanol-water（72:28）, for Ros A was methanol-0.1% trifluoroacetic acid（TFA） with gradient elution（0～5 min, 35%B, 5.01～15 min, 35%～40%B, 15～30 min, 40%B）. The UV detection wavelength for SSb was 254 nm, for RosA was 330 nm. The flow rate was 1.0 mL·min-1, and the column temperature was 35 ℃. Results The linear ranges of SSb1, b2 and RosA were 0.37～9.25 μg（r=0.999 9）, 0.52～13 μg（r=0.999 9）, 0.2～5.61 μg（r=0.999 9）. The average recovery was 96.89%, 99.01%, 97.09%, with RSD of 1.88%, 2.28%, 1.65%（n=6）. Conclusion: The method is accurate, simple and has a well reproduci bility, which can be used as principles of quantitative analysis for Jiayan Kangtai Granules.