红芪精准煮散饮片HPLC指纹图谱建立及3种指标成分测定

Establishment of HPLC fingerprint of Hedysari Radix precision boiled powder decoction pieces and determination of three indicative components

目的建立红芪精准煮散饮片高效液相色谱指纹图谱和3种指标成分的含量测定方法。方法采用HPLC法测定红芪饮片与红芪精准煮散饮片中芒柄花苷、毛蕊异黄酮和芒柄花素的含量,色谱柱为Robusta C_(18)柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.2%磷酸水溶液,梯度洗脱,检测波长为254 nm;体积流量1.0 mL/min;进样量20μL;柱温30℃。结果建立了红芪精准煮散饮片的特征指纹图谱,并指认出了20个共有峰中的3个主要峰芒柄花苷(峰11)、毛蕊异黄酮(峰13)和芒柄花素(峰16),3种成分在各自范围内线性关系良好(r≥0.9995),平均加样回收率98.08%~99.33%,RSD在1.46%~2.37%。通过加权得分,比较红芪精准煮散饮片与传统饮片中3种成分,得到红芪精准煮散饮片1.00 g约相当于传统饮片1.37 g。结论建立的指纹图谱及含量测定方法可为红芪精准煮散饮片的质量控制与评价提供依据。

Objective To establish the HPLC fingerprint and the content determination method of the three indicative components of Hongqi(Hedysari Radix)precision boiled powder decoction pieces.Methods HPLC was used to determine the contents of ononin,calycosin and formononetin in Hedysari Radix traditional decoction pieces and Hedysari Radix precision boiled powder decoction pieces.The chromatographic column was Robusta C_(18)(250 mm×4.6 mm,5μm),the mobile phase was acetonitrile-0.2%phosphoric acid solution,and the gradient elution was used.The detection wavelength was 254 nm;The flow rate is 1.0 mL/min;20μL of sample injection;Column temperature is 30℃.Results The characteristic fingerprint of Hedysari Radix precision boiled powder decoction pieces was established,and three main peaks of 20 common peaks,namely ononin(peak 11),calycosin(peak 13)and formononetin(peak 16),were identified.The linear relationship between the three components was good within their respective ranges(r≥0.9995),with the average recovery rate of 98.08%—99.33%and RSD of 1.46%—2.37%.Through the weighted score,comparing these three components in the Hedysari Radix precision boiled powder decoction pieces with the traditional decoction pieces,it is found that 1 g of Hedysari Radix precision boiled powder decoction pieces is about 1.37 g of the traditional decoction pieces.Conclusion The established fingerprint and content determination method can provide the basis for the quality control and evaluation of Hedysari Radix precision boiled powder decoction pieces.