目的 建立2种测定牛黄上清制剂中4种成分的HPLC方法,采用双标线性校正法进行研究,预测保留时间,考察其准确性和色谱柱的符合率,用于辅助色谱峰定性。方法 选用20根不同品牌和型号的C18色谱柱,测定牛黄上清制剂中4种成分的实际保留时间,计算平均值,得到标准保留时间(standard retention time,ts),以峰1栀子苷和峰4黄芩苷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,并在其他品牌和型号的5根C18色谱柱上进行方法验证。同时,以中间峰芍药苷为参照物,计算校正因子,使用相对保留时间法预测保留时间。将双标线性校正法和相对保留时间法两种替代对照品定性方法进行优劣比较。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。
Abstract
OBJECTIVE To establish an analysis method for determination of four components in two kinds of Niuhuangshangqing preparations by HPLC and predicte the retention time by doable standard linear calibration method, in which the retention time was predicted by double standard linear calibration method and the prediction accuracy and column suitability were investigated in order to assist the chromatographic peak identification. METHODS The retention time of the four components in Niuhuangshangqing preparations were determined on 20 C18 columns of different brands and types.The standard retention time (ts) was calculated. Both geniposide (peak 1) and baicalin (peak 4) were taken as the linear calibration standards to forecast the retention time of the other two components. Other five C18 columns of different brands and types were used for validation. In comparison of liner calibration with the two reference substances,paeoniflorin a was used as the reference substance for the determination of relative correction factors of other three 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, which is demonstrated to be practical and promising for multi-component assay of traditional Chinese medicine.
关键词
双标线性校正法 /
色谱峰定性 /
牛黄上清制剂 /
多指标成分
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Key words
liner calibration with two reference substances /
chromatographic peak identification /
Niuhuangshangqing preparation /
multiple component
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中图分类号：
R917
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参考文献
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