牛黄上清丸中多指标成分含量测定方法研究

Determination of Geniposid,Baicalin,Sennoside A and Sennoside B in Niuhuangshangqing Pill Using HPLC-DAD

目的:建立牛黄上清丸中同时测定桅子苷、黄芩苷、番泻苷A和番泻苷B四个指标成分含量的方法。方法:色谱柱为AgilentZorbax SB-C_(18)(5μm,250×4.6 mm),流动相为乙腈-0.05%磷酸水梯度洗脱,流速为1ml/min,检测波长280nm。结果:栀子苷、黄芩苷、番泻苷A和番泻苷B的分离度良好(R>1.5),四条标准曲线在检测范围内均呈良好线性(r≥0.9999),其检测限均低于2.90ng、定量限均低于6.70ng,精密度的RSD均小于0.97%,加样回收率分别高于为96.1%、97.2%、94.5%和92.6%。结论:该方法简便、快速、灵敏、准确,在同一色谱检测条件下实现多指标成分同时定量,为该药的全面质量评价提供参考。

Objective： To establish the method for simultaneous determination of geniposid, baicalin, sennoside A and sennoside B in Niuhuangshangqing pill using HPLC-DAD. Methods：The analysis was achieved with an Agilent Zorbax SB-C^18 analytical column （250x 4.6 mm I.D., 5μm） by phosphoric acid in water and acatonitrile. The flow rate was 1 ml min-l, and detection wavelength was set at 280nm. Results：Good resolutions of the four determining peaks with their adjacent peaks were discovered （R〉 1.5）. The four calibration curves showed good linearity （r≥0.9999）. The LOD and LOQ were lower than 2.90 ng and 6.70 ng on column, respectively. The precision RSD of the four analytes was less than 0.97% at three levels, and the recoveries were higher than 96.1%, 97.2%, 94.5% and 92.6% respectively. Conclusions： The developed HPLC assay is accurate and reliable, and could be readily utilized as a determination method of Niuhuangshangqing pill. This method could provide reference for comprehensive quality evaluation.