黄芪颗粒的HPLC特征图谱研究及4个成分含量测定

目的: 建立黄芪颗粒的HPLC特征指纹图谱分析方法,并测定该制剂中毛蕊异黄酮苷、芒柄花苷、毛蕊异黄酮、芒柄花素的含量,为其质量控制提供依据。 方法: 利用HPLC法,采用Agilent ZORBAX SB-C₁₈色谱柱(5 μm,4.6 mm×250 mm),使用甲醇-0.5%甲酸水溶液为流动相进行梯度洗脱,检测波长254 nm。采用外标法,利用毛蕊异黄酮苷、芒柄花苷、毛蕊异黄酮、芒柄花素的混合对照测定其在30批制剂中的含量。 结果: 建立了黄芪颗粒HPLC特征指纹图谱共有模式,标定了23个共有峰,指认了其中4个共有峰,统计结果显示,不同生产时间30批次黄芪颗粒被分为两大类,两类样品的色谱峰共有模式具有一定差异,这与黄芪颗粒投料原料黄芪药材有2个法定来源相吻合。4个物质在两类制剂之间含量存在较大差异,两类制剂之内含量基本一致。 结论: HPLC特征图谱方法与4个物质的含量测定方法快速、准确、简便,均可用于黄芪颗粒的质量控制。

Objective: To establish an HPLC method for fingerprint analysis of Huangqi granules and determination of the contents such as calycosin-7-O-β-D-glucoside,ononin,calycosin and formononetin,so as to provide a scientific basis for the quality control. Methods: Separation was performed on an Agilent ZORBAX SB-C₁₈ column(5 μm,4.6 mm×250 mm).The gradient elution was performed by the mobile phase consisting of methanol and 0.5% formic acid aqueous with the flow rate of 1 mL·min^-1,the detection wavelength was set at 254 nm,and the column temperature was 30℃.The mixed reference substances for calycosin-7-O-β-D-glucoside,ononin,calycosin and formononetin were used to determine their contents in 30 batches of formulations by the external standard method. Results: The common pattern of the HPLC characteristic chromatographic profile for Huangqi granules was set up.There were 23 common peaks marked,four of which were identified.Mathematical statistics results showed that 30 batches of Huangqi granules were divided into two categories,and the mutual model of two kinds of samples had a certain difference,which was consistent with the fact that the raw materials of Huangqi granules had 2 legal sources.There were relatively larger differences in the contents of the four substances between the 2 formulations,but the contents were basically the same within each formulation. Conclusion: The method is accurate,and simple,which can be used for quality control of Huangqi granules.