基于双标线性校正法的银黄口服液多组分定性分析

Qualitative determination of multiple components in Yinhuang oral liquid by linear calibration using two reference substances

目的:建立银黄口服液中7个成分(新绿原酸、绿原酸、隐绿原酸、3,4-O-二咖啡酰奎宁酸、3,5-O-二咖啡酰奎宁酸、4,5-O-二咖啡酰奎宁酸、黄芩苷)的高效液相色谱方法,考察双标线性校正法用于色谱峰定性的可行性,并比较双标线性校正法和相对保留时间法2种替代对照品定性方法的优劣。方法:采用25根不同品牌或型号的C18色谱柱上,测定银黄口服液中7个成分的实际保留时间,并计算标准保留时间(SRT)。借助DRS Origin软件,以新绿原酸(峰1)和黄芩苷(峰7)为双标化合物,采用双标线性校正法预测其他5个成分的保留时间,并在另一根C18色谱柱上进行方法学验证。同时以绿原酸(峰2)为参照物,使用相对保留时间法预测其他6个成分的保留时间。结果:双标线性校正法可用于银黄口服液的多组分定性分析,且建立的方法符合方法学验证要求。与相对保留时间法相比,双标线性校正法预测保留时间的准确度更高,色谱柱的适用范围更广。结论:双标线性校正法能较好地辅助色谱峰定性,方法可行、准确且经济实用。

Objective:To establish an HPLC method of Yinhuang oral liquid for determination of seven components(neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,3,4-O-dicaffeoyl quinic acid,3,5-O-dicaffeoyl quinic acid,4,5-O-dicaffeoyl quinic acid and baicalin),in order to detect the feasibility of linear calibration method using two reference substances.And the two alternative qualitative methods of reference were compared between linear calibration method using two reference substances and the relative retention time method.Methods:The retention time of the seven components in Yinhuang oral liquid were determined on 25 C18 columns of different brands or types and the standard retention time(SRT)was calculated.Both neochlorogenic acid(peak 1)and baicalin(peak 7)were taken as the two reference substances to forecast the retention time of the other five components by linear calibration method using two reference substances,with the aid of DRS origin software.Another C18 column was used for methodology validation.Chlorogenic acid(peak 2)was used as the reference substance to forecast the retention time of the other six components by relative retention time method.Results:Linaer calibration using two reference substances could be used to the qualitative determination of multiple components in Yinhuang oral liquid.The established method met the requirements of methodology validation.Linear calibration using two reference substances was more accurate and suitable for more chromatographic columns than relative retention time method.Conclusion:Linear calibration using two reference substances can be used to assist the chromatographic peak identification,and was feasible,reliable and economical.