双标线性校正法用于川黄芩的多成分定性分析

Multicomponent Qualitative Analysis of Scutellariae Amoenae Radix by Linear Calibration Using Two Reference Substances

目的 建立测定川黄芩中5个成分(黄芩苷、未知峰2、千层纸素A-7-O-β-D-葡萄糖醛酸苷、汉黄芩苷、黄芩素)的高效液相色谱(HPLC)方法,并比较双标线性校正法和相对保留时间法的优劣。方法 采用20根不同品牌的C_(18)色谱柱,测定川黄芩中5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,以黄芩苷(峰1)和黄芩素(峰5)为双标化合物,采用双标线性校正法预测其他3个成分的保留时间,并在另外两根C_(18)色谱柱上进行方法验证。同时以千层纸素A-7-O-β-D-葡萄糖醛酸苷(峰3)为参照物,使用相对保留时间法预测其他4个成分的保留时间。结果 双标线性校正法可用于川黄芩的多成分定性分析,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 双标线性校正法具有更好的辅助色谱峰定性的作用,方法可行、准确且经济实用。

Objective To establish an HPLC method for determination of five components in Scutellariae Amoenae Radix(baicalin,unknown peak 2,oroxylin A-7-O-β-D-glucoside,wogonoside and baicalein),and compare the advantages and disadvantages of linear calibration using two reference substances(LCTRS)and relative retention time method.Methods Twenty C_(18)columns of different brands were used to determine the actual retention time of five components in Scutellariae Amoenae Radix and calculate the standard retention time(SRT).With the aid of DRS Origin software,baicalin(peak 1)and baicalein(peak 5)were used as two reference substances,and the retention time of the other three components was predicted by LCTRS,and the method was validated on other two C_(18) chromatographic columns.At the same time,using oroxylin A-7-O-β-D-glucoside(peak 3)as the reference,the retention time of the other four components was predicted by relative retention time method.Results LCTRS could be used for the qualitative analysis of multiple components in Scutellariae Amoenae Radix.Compared with the relative retention time method,LCTRS had higher accuracy in predicting retention time and wider application range of chromatographic columns.Conclusion LCTRS is feasible,accurate and economical,which is more effective in assisting the qualitative determination of chromatographic peaks.