双标线性校正法用于一清颗粒的多指标成分定性分析

Determination of Multiple Components in Yiqing Granules by Liner Calibration with Two Reference Substances

目的建立检测清颗粒中8种成分的高效液相色谱法,采用双标线性校正法计算保留时间,考察其准确性和色谱柱的符合率,用于色谱峰定性分析。方法选用24根不同品牌和型号的C色谱柱,测定一清颗粒中8种成分的实际保留时间并计算平均值,得到标准保留时间(t),以峰1表小檗碱和峰8汉黄芩素两个成分作为双标化合物,使用双标线性校正法预测保留时间,并在4根其他品牌和型号的C色谱柱上进行方法验证。同时,以中间峰盐酸巴马汀为参照物,计算校正因子,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的优劣。结果双标线性校正法与相对保留时间法相比,具有更高的保留时间预测正确率和色谱柱符合率,能适用于更多的色谱柱。结论双标线性校正法能较好地辅助色谱峰定性,在中药多指标成分分析方面有较好的实用价值和应用前景。

OBJECTIVE To establish an analysis method of Yiqing Granules for determination of eight components by HPLC chromatographic and predict the retention time by liner calibration with two reference substances.METHODS The retention time of the eight components in Yiqing Granules was determined on 24 C18 columns of different brands and types.The standard retention time(t)was calculated.Both epiberberine(peak 1)and wogonin(peak 8)were taken as the linear calibration standards to forecast the retention time of the other six components.Other four C18 columns of different brands and types were used for validation.In comparison of liner calibration with the two reference substances,palmatine hydrochloride was used as the reference substance for the determination of relative correction factors of the other seven components.RESULTS Liner calibration with two reference substances was more accurate for predicting the retention time and suitable for more kinds of chromatographic columns than the relative retention time method.CONCLUSION The method of liner calibration with two reference substances can be used to assist the chromatographic peak identification,with a high practical value and application prospect for multi-component assay of traditional Chinese medicine.