基于HPLC指纹图谱结合化学模式识别的护肝片质量控制研究

Quality evaluation of Hugan Tablets based on HPLC fingerprint and chemical pattern recognition

目的建立中成药护肝片的HPLC指纹图谱,为其质量评价提供依据。方法采用HPLC法,以五味子醇甲为参照,绘制10批护肝片原料粉样品的HPLC图谱;利用中药色谱指纹图谱相似度评价系统(2004 A)进行相似度评价,确定共有峰。采用主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)对样品进行模式识别分析。结果 10批护肝片样品的HPLC图谱有22个共有峰,经对照品进行化学指认共鉴定了其中的6个色谱峰,分别是五味子醇甲、柴胡皂苷a、柴胡皂苷c、柴胡皂苷d、五味子甲素、五味子乙素。10批供试品的相似度均大于0.97,表明该药物总体质量较为稳定;但通过PCA发现不同批次药物质量之间存在微小差异,且主要分为2类,最后进一步采用OPLS-DA筛选出了导致批次药物质量差异的2种主要成分,分别为12号峰(柴胡皂苷d)和22号峰。结论护肝片指纹图谱的构建和化学模式识别为护肝片的质量评价提供了可行的理论依据,可为生产厂家在药品生产中更高效、合理地控制药品质量提供科学依据。

Objective To establish the HPLC fingerprint of Hugan Tablets and provide a scientific basis for the quality control of Hugan Tablets. Methods HPLC method was adopted and using schizandrin as reference, and 10 batches of Hugan Tablets were determined. Similarity evaluation was performed by using Similarity Evaluation System for Fingerprint Chromatogram of TCM (2004A) to confirm the common peak. Pattern recognition analysis was performed using principal component analysis (PCA) and orthogonal partial least squares-discrimination analysis (OPLS-DA). Results There were 22 common peaks in HPLC of 10 batches of samples. Schizandrin, saikosaponin a, saikosaponin c, saikosaponin d, schisandrin A, and schizandrin B were identified by chemical identification of the reference substance. The similarity of 10 batches of samples was higher than 0.97, which indicated that the overall quality of the drug was relatively stable. However, there were slight differences between the quality of different batches of drugs found by PCA, and they were mainly divided into two categories. Finally, OPLS-DA was used to screen out two main components that caused the quality differences in the batches, namely peak 12 (saikosaponin D) and peak 22. Conclusion The construction of fingerprint of Hugan Tablets and chemical pattern recognition provides a practical and theoretical basis for the quality evaluation of Hugan Tablets, which can provide a scientific basis for manufacturers to control drug quality more efficiently and reasonably in drug production.