基于UHPLC-Q-exactive orbitrap MS和化学计量学分析的芪龙壮儿口服液质量评价研究

Quality evaluation of Qilong Zhuang'er oral liquid based on UHPLC-Q-exactive orbitrap MS and chemometrics analysis

建立同时测定芪龙壮儿口服液中19种化学成分的UHPLC-Q-exactive orbitrap MS定量分析方法,并结合化学计量学分析比较不同批次样品的质量差异,为该制剂的质量评价提供依据。采用UHPLC-Q-exactive orbitrap MS方法测定了10批芪龙壮儿口服液中尿囊素、L-脯氨酸、焦谷氨酸、大麦芽碱、腺苷、L-苯丙氨酸、鸟苷、L-色氨酸、咖啡酸、毛蕊异黄酮苷、毛蕊花糖苷、异毛蕊花糖苷、芒柄花苷、毛蕊异黄酮、3-羟基-9,10-二甲氧基紫檀烷、芒柄花素、白术内酯Ⅲ、白术内酯Ⅱ和黄芪甲苷的含量,并利用Hiplot开源科研绘图平台及MarkerlynxXS软件对含量数据进行聚类热图分析、主成分分析(principal component analysis, PCA)和偏最小二乘判别分析(partial least squares discriminant analysis, PLS-DA),综合评价不同批次芪龙壮儿口服液的质量差异。19个化学成分在各自浓度范围内线性关系良好(r≥0.999),精密度、重复性、稳定性(24 h)试验的RSD均小于1.94%,平均加样回收率为97.24%~102.75%(RSD均小于2.74%, n=6)。聚类热图和PCA分析将10批样品分为两大类, PLS-DA法确定了7个差异标志物,分别为3-羟基-9,10-二甲氧基紫檀烷、白术内酯Ⅲ、毛蕊异黄酮、白术内酯Ⅱ、芒柄花素、尿囊素和咖啡酸。建立的芪龙壮儿口服液多成分定量方法结合化学计量学分析可以为该制剂质量评价提供参考。

An UHPLC-Q-exactive orbitrap MS method for the simultaneous determination of 19 chemical components in Qilong Zhuang'er oral liquid was established and the quality differences between different batches of samples was compared by chemometric analysis to provide a basis for the quality evaluation of the preparation.The contents of allantoin,L-proline,pyroglutamic acid,hordenine,adenosine,L-phenylalanine,guanosine,L-tryptophan,caffeic acid,calycosin-7-glucoside,verbascoside,isoacteoside,ononin,calycosin,3-hydroxy-9,10-dimethoxyptercarpan,formononetin,atractylenolideⅢ,atractylenolideⅡand astragaloside A were analyzed by cluster heat map,principal component analysis(PCA)and partial least squares discriminant analysis(PLS-DA)using Hiplot platform and MarkerlynxXS software to comprehensively evaluate the quality difference of different batches of Qilong Zhuang'er oral liquid.The 19 chemical compounds showed good linearity in their respective concentration ranges(r≥0.999).The RSD of precision,repeatability and stability(24 h)tests were all less than 1.94%.The average recovery was 97.24%-102.75%(RSD<2.74%,n=6).The 10 batches of samples were divided into two categories by cluster heat map and PCA analysis.3-Hydroxy-9,10-dimethoxyptercarpan,atractylenolideⅢ,calycosin,atractylenolideⅡ,formononetin,allantoin and caffeic acid were identified as differential markers by PLS-DA.The established multi component quantitative method of Qilong Zhuang'er oral liquid combined with chemometric analysis can provide reference for the quality evaluation of the preparation.