双标线性校正法辅助色谱峰定性用于双黄连制剂的多组分分析

Determination of Multiple Components in Shuanghuanglian Preparations by Liner Calibration with Two Reference Substances Assisted with Chromatographic Peaks Identification

目的：建立4种常用双黄连制剂中9种成分的高效液相色谱方法,采用双标线性校正法计算保留时间,考察预测准确性和色谱柱符合率,以辅助色谱峰定性分析。方法：选用28根不同品牌和型号的C18色谱柱,测定双黄连制剂中9种成分的实际保留时间,计算平均值得到标准保留时间（SRT）,以峰1绿原酸和峰9汉黄芩素2个成分为双标化合物,使用双标线性校正法预测保留时间,并在5根其他品牌和型号的C18色谱柱上进行了方法验证。同时以中间峰黄芩苷为参照物,计算校正因子,使用相对保留时间法预测保留时间。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。结果：与相对保留时间法相比,双标线性校正法具有更高的保留时间预测正确率和色谱柱符合率,能够适用于更多的色谱柱。但其在标准化和实用性方面,仍需继续深入研究。能否推广到其他品种,也需实验论证。结论：双标线性校正法能较好地辅助色谱峰定性,是一种新的替代对照品方法,具有广阔的应用前景。

Objective： To establish an analysis method of Shuanghuanglian preparations for determination of multiple components by HPLC chromatographic,in order to assist the peak identification,and detect the prediction accuracy and column suitability. Method： The retention time of the nine components in Shuanghuanglian preparations were determined on 28 C18 columns of different brands and types. The standard retention time（SRT） was calculated. Both chlorogenic acid（peak 1） and wogonin（peak 9） were taken as the linear calibration standards to forecast the retention time of the other seven components. Other 5 C18 columns of different brands and types were used for validation. In comparison of liner calibration with the two reference substances,baicalin was used as the reference substance for the determination of relative correction factors of other eight components. Result： Liner calibration with two reference substances is more accurate for predicting the retention time and suitable for more kinds of chromatographic columns than the relative retention time method.Further studies shall be made for the standardization and practicability. Conclusion： The method of liner calibration with two reference substances can be used to assist the chromatographic peak identification,with a broad practical prospect.