基于高效液相色谱法的蒲公英指纹图谱研究

Establishment of Taraxaci Herba fingerprint based on high performance liquid chromatography

本试验旨在建立蒲公英的高效液相色谱指纹图谱,并将其应用于不同批次的蒲公英样品检测。采用Waters Atlantis®T3(250.0 mm×4.6 mm,5.0µm)色谱柱,流动相为0.1%甲酸溶液(含0.05%三乙胺)-乙腈,梯度洗脱;检测波长为325 nm;流速为1 mL/min;柱温30℃。方法学考察合格后,检测15批蒲公英样品,用中药色谱指纹图谱相似度评价系统建立指纹图谱,进行相似度评价;以保留时间和最大吸收波长为参考标准,鉴定色谱峰;用SPSS 20.0软件进行聚类分析和主成分分析。结果显示,本试验成功建立了蒲公英的高效液相色谱指纹图谱,15批蒲公英样品的相似度为0.611~0.999,共标定出15个共有峰,通过对照品指认了其中5个峰,分别为单咖啡酰酒石酸、绿原酸、咖啡酸、菊苣酸和异绿原酸A。聚类分析将15批蒲公英样品分为3类,主成分分析后15个共有峰被分为5个主成分。本试验对样品中各色谱峰的光谱吸收特点进行了比对,与现有其他方法比较,该方法快速可靠,稳定性和重复性良好,可为蒲公英的质量评价提供参考。

The objective of this study was to establish the high-performance liquid chromatography fingerprint of Taraxaci Herba and apply it to the detection of different batches of Taraxaci Herba samples.The chromatographic conditions were as follows:Waters Atlantis®T3(250.0 mm×4.6 mm,5.0µm)column was used;the mobile phase was 0.1%formic acid aqueous solution containing 0.05%triethylamine-acetonitrile with gradient elution;the detection wavelength was 325 nm;flow rate of 1 mL/min;the column temperature was 30℃.After the methodological investigation was qualified,15 batches of Taraxaci Herba samples were tested,and the fingerprint was established by the traditional Chinese medicine chromatographic fingerprint similarity evaluation system for similarity evaluation.Chromatographic peaks were identified using retention time and maximum absorption wavelength as reference standards;SPSS 20.0 software was used for cluster analysis and principal component analysis.The results showed that the high-performance liquid chromatography fingerprint of Taraxaci Herba was established,and the similarity of 15 batches of Taraxaci Herba samples was between 0.611-0.999,and a total of 15 common peaks were calibrated,and 5 of them were identified by the reference substance,which were monocaffeoyl tartaric acid,chlorogenic acid,caffeic acid,chichoric acid and isochlorogenic acid A.15 batches of Taraxaci Herba samples were divided into 3 categories by cluster analysis,and the 15 common peaks were divided into 5 principal components after principal component analysis.Compared with other existing methods,the spectral absorption characteristics of each chromatographic peak in the sample were compared in this experiment,which showed that the method was fast,reliable,stable and repeatable,and could provide a reference for the quality evaluation of Taraxaci Herba.