基于超高效液相色谱串联电喷雾式检测器新技术的熊胆粉特征指纹图谱研究

Study on characteristic fingerprint of bear bile powder based on ultra-high performance liquid chromatography tandem charged aerosol detector

目的:解决熊胆粉质量控制存在的胆汁酸类化合物检测困难、化学信息不足、耗时长等问题,为熊胆粉质量控制提供新的思路和方法。方法:采用超高效液相色谱串联电喷雾式检测器(UPLC-CAD)新技术,建立熊胆粉特征指纹图谱分析方法,并利用相似度计算方法结合主成分分析(PCA)及偏最小二乘法-判别分析(PLS-DA)对35批不同来源熊胆粉进行评价。结果:建立的UPLC-CAD分析方法具有良好的精密度和重复性,结合液质联用对8个共有峰进行化学指认,全面地反映了熊胆粉中的主要胆汁酸成分。不同方法相似度评价结果显示,投影含量法可以更好地反映化学成分含量相似度。PCA和PLS-DA分析结果显示,不同来源熊胆粉可聚集为3类,熊胆粉主要活性成分牛磺熊去氧胆酸(TUDCA)和牛磺鹅去氧胆酸(TCDCA)在不同来源样品中具有较好的一致性,而差异显著成分为其他含量较小的胆汁酸成分如7-酮-牛磺石胆酸、胆酸等,提示熊胆粉质量控制应以主要指标成分TUDCA和TCDCA含量结合胆汁酸特征指纹图谱进行综合评价。结论:本研究所建立的基于UPLC-CAD的熊胆粉特征指纹图谱分析方法简便快捷,可为熊胆粉的质量标准及药效物质基础研究提供依据,并为其他含弱紫外吸收化学成分的中药材质量控制提供参考。

Objective: To solve the problems in quality control of bear bile powder, such as difficult detection of bile acid compounds, lack of chemical information, and long time-consuming, and provide new ideas and methods for quality control of bear bile powder. Methods: A new method of ultra-high performance liquid chromatography tandem charged aerosol detector(UPLC-CAD) was established to analyze thecharacteristic fingerprint of bear bile powder. In addition, 35 batches of bear bile powder from different sources were evaluated by similarity calculation methods, principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA). Results: The established UPLC-CAD method showed good precision and repeatability. Combined with ultra-high performance liquid chromatography-mass spectrometry technology, the structures of eight common peaks were identified, which comprehensively reflected theprincipal bile acids in bear bile powder. The similarity evaluation results of different methods showed that the projection content method could better reflect the similarity of chemical composition content. Further,PCA and PLS-DA showed that bear bile powder fromdifferent sources could be grouped into three categories. Themain active components,tauroursodeoxycholic acid( TUDCA) and taurochendeoxycholalic acid( TCDCA),showed good consistency in of bear bile powder fromdifferent sources, while 7-ketotaurolithocholic acid,cholic acidand other bile acids with low contents were significantly different. It was suggested that the quality control of bear bile powder should integratethecontent of main index components TUDCA and TCDCA with the characteristic fingerprint of bile acids. Conclusion: In this study,a simple and rapid UPLC-CAD method is established for analyzing characteristic fingerprint of bear bile powder,which can provide basis for quality control and pharmacodynamic material basis study,and also provide reference for quality control of other traditional Chinese medicines without strong ultraviolet absorption.