摘要
目的 建立HPLC波长切换法同时测定通络治痹方中绿原酸、羟基红花黄色素A、阿魏酸、山柰酚3-O-芸香糖苷、蛇床子素、二氢欧山芹醇当归酸酯6种有效成分含量.方法 色谱柱为XBridge C18柱(250 mm×4.6 mm,5 µm),流动相为乙腈(A)-0.1％磷酸水(B),梯度洗脱,流速1 ml/min,柱温30 ℃,检测波长330 nm(0～14 min检测绿原酸,15～80 min检测阿魏酸、山柰酚3-O-芸香糖苷、蛇床子素、二氢欧山芹醇当归酸酯)、403 nm(14～15 min检测羟基红花黄色素A).结果 绿原酸、羟基红黄花色素A、阿魏酸、山柰酚3-O-芸香糖苷、蛇床子素、二氢欧山芹醇当归酸酯分别在0.029 7～1.485 0、0.030 0～1.500 0、0.009 9～0.495 0、0.017 5～0.875 0、0.028 4～1.420 0、0.013 7～0.685 0 μg内具有良好线性关系(R2≥0.999 8),精密度、稳定性(24h)、重复性RSD均低于2％.平均加样回收率在95％～105％,RSD均在0.32％～1.67％.10批通络治痹方供试样品中,上述6个成分含量测定结果依次为0.221 60、0.314 30、0.085 10、0.032 95、0.043 87、0.026 21 mg/g.结论 建立的 HPLC波长切换法快速、简便而准确,可同时测定通络治痹方中上述6种成分含量,为通络治痹方质量控制及新剂型研究提供参考.
Abstract
Objective To establish high performance liquid chromatography(HPLC)wavelength switching method to simultaneously determine the contents of chlorogenic acid,hydroxysafflor yellow A,ferulic acid,Nicotiflorin,Osthole and columbianadin in Tongluo Zhibi Prescription.Methods The column was XBridge C18 column(250 mm × 4.6 mm,5 μm);the mobile phase was acetonitrile(A)-0.1％phosphate water(B);gradient eluted,with flow rate:1 ml/min,column temperature:30 ℃,detection wavelength 330 nm(0-14 min detection of chlorogenic acid,15-80 min detection of ferulic acid,Nicotiflorin,Osthole,and columbianadin),403 nm(14-15 min detection of hydroxysafflower yellow pigment A).Results Chlorogenic acid,hydroxyrhodopsin A,ferulic acid,kaempferol 3-O-rutinoside,serpentin,and dihydroeurobicarpus angelicus acid ester showed good linearity(R2 ≥ 0.999 8)within 0.029 7-1.485 0,0.030 0-1.500 0,0.009 9-0.495 0,0.017 5-0.875 0,0.028 4-1.420 0,0.013 7-0.685 0 μg,respectively.The precision,stability(24 h),repeatability relative standard deviation(RSD)were all＜2％.The average spiked recoveries were all in the range of 95％-105％,and the RSDs were all in the range of 0.32％-1.67％.In 10 batches of test samples of Tongluo Zhibi Prescription,the content of the above six components,including chlorogenic acid,was determined to be 0.221 60,0.314 30,0.085 10,0.032 95,0.043 87,0.026 21 mg/g in the following order.Conclusion The established HPLC wavelength switching method is fast,simple and accurate,which can be used for simultaneous determination of the content of the above six components in Tongluo Zhibi Prescription,which provides reference for quality monitoring and new dosage form research of Tongluo Zhibi Prescription.
基金项目
江苏省中医退行性骨关节病临床医学创新中心项目(苏中医科教[2021]4号)
NETL
NSTL
万方数据