基于HPLC特征图谱、多成分含量测定与化学模式识别的气血康口服液质量研究

Quality Study of Qixuekang Oral Liquid Based on HPLC Characteristic Chromatogram, Multi-Component Content Determination, and Chemical Pattern Recognition

中文关键词:气血康口服液  HPLC特征图谱  含量测定  化学模式识别  质量研究

keywords:Qixuekang Oral Liquid  HPLC characteristic chromatogram  content determination  chemical pattern recognition  quality study

中文摘要:［摘要］目的：建立气血康口服液的高效液相色谱法（HPLC）特征图谱、多成分含量测定方法，并结合化学模式识别分析气血康口服液的质量。方法：采用HPLC-紫外法（UV）、HPLC-蒸发光散射检测法（ELSD）建立24批气血康口服液的HPLC特征图谱，并测定其含有的6个成分（黄芪甲苷、毛蕊异黄酮葡萄糖苷、人参皂苷Rb1、人参皂苷Rg1、三七皂苷R1、葛根素）的含量，结合化学模式识别筛选影响其质量的关键成分。结果：基于HPLC-UV建立的气血康口服液HPLC特征图谱中共确定了24个共有峰，24批气血康口服液的HPLC特征图谱与其对照特征图谱间的相似度均大于0.96；基于HPLC-ELSD建立的HPLC特征图谱中共确定了10个共有峰，以峰4为参照峰，9个共有峰相对保留时间的RSD为0.16%~0.61%。经与对照品比对，共指认出6个成分，分别为黄芪甲苷、毛蕊异黄酮葡萄糖苷、人参皂苷Rb1、人参皂苷Rg1、三七皂苷R1、葛根素。上述6个成分含量测定的线性关系考察结果显示，6个成分的质量浓度在其相应范围内线性关系良好；精密度试验、稳定性试验结果表明，6个成分峰面积、峰面积对数（ln）的RSD均小于3.2%；重复性试验结果表明，6个成分含量的RSD均小于3.0%；加样回收率结果表明，6个成分加样回收率为97.72%~105.40%，RSD为0.42%~4.00%。聚类分析（HCA）、主成分分析（PCA）结果表明24批气血康口服液聚为2类；正交偏最小二乘法-判别分析（OPLS-DA）表明，基于变量重要性投影（VIP）值>1筛选出对质量影响较大的3个关键成分（人参皂苷Rg1、人参皂苷Rb1、三七皂苷R1）。结论：建立的HPLC特征图谱和多成分含量测定方法简便准确，结合化学模式识别可为气血康口服液的质量控制提供参考。

Abstract:[Abstract] Objective: This paper aims to establish the high-performance liquid chromatography (HPLC) characteristic chromatogram and a multi-component content determination method for Qixuekang Oral Liquid and to evaluate its quality using chemical pattern recognition. Methods: HPLC-ultraviolet (UV) and HPLC-evaporative light scattering detection (ELSD) methods were used to establish the HPLC characteristic chromatograms of 24 batches of Qixuekang Oral Liquid. The contents of six components, including astragaloside IV, calycosin-7-O-β-D-glucoside, ginsenoside Rb1, ginsenoside Rg1, notoginsenoside R1, and puerarin, were simultaneously determined. Chemical pattern recognition was further applied to screen the key components affecting its quality. Results: A total of 24 common peaks were identified in the HPLC characteristic chromatogram of Qixuekang Oral Liquid established by HPLC-UV, and the similarities between the HPLC characteristic chromatograms of 24 batches of Qixuekang Oral Liquid and the reference characteristic chromatogram were all higher than 0.96. Ten common peaks were confirmed in the HPLC characteristic chromatograms established by HPLC-ELSD. With peak 4 set as the reference peak, the relative standard deviations (RSDs) of the relative retention time of nine common peaks ranged from 0.16% to 0.61%. Six components were identified by comparison with reference samples, namely astragaloside Ⅳ, calycosin-7-O-β-D-glucoside, ginsenoside Rb1, ginsenoside Rg1, notoginsenoside R1, and puerarin. The results of the linear relationship investigation for the determination of the above six components show that good linear relationships exist among mass concentrations of the six components within their corresponding concentration ranges. Precision and stability tests show that the RSDs of both peak areas and their logarithm (ln) for the six components are all less than 3.2%. Repeatability tests indicate that the RSDs of the six component contents are all less than 3.0%. The results show that spike recoveries of the six components range from 97.72% to 105.40%, with RSDs of 0.42%–4.00%. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) results show that 24 batches of Qixuekang Oral Liquid are divided into two categories. Orthogonal partial least squares-discriminant analysis (OPLS-DA) results show that three key components exerting major influences on quality (ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1) are screened out, based on variable importance in projection (VIP) value greater than 1. Conclusion: The established HPLC characteristic chromatograms and multi-component content determination method in this study are simple and accurate. Combined with chemical pattern recognition, they can provide a reference for the quality control of Qixuekang Oral Liquid.