基于指标成分测定及指纹图谱模式识别的紫苏叶产地差异性研究

Regional differences of Perillae Folium based on index component determination and fingerprint pattern recognition

目的对不同产地的紫苏Perilla frutescens叶进行研究,建立紫苏叶的指纹图谱并测定野黄芩苷、迷迭香酸含量,为紫苏叶的质量控制提供参考。方法收集9个产地共36批紫苏叶药材,以乙腈-0.2%甲酸溶液为流动相,建立紫苏叶中野黄芩苷、迷迭香酸的HPLC含量测定方法,对36批紫苏叶进行含量测定,同时建立紫苏叶指纹图谱并进行相似度评价;采用微科盟生科云、SIMCA 14.1等工具进行聚类分析(cluster analysis,CA)、主成分分析(principal component analysis,PCA)、偏最小二乘法判别分析(partial least squares discriminant analysis,PLS-DA)。结果以野黄芩苷为参照峰建立了紫苏叶的指纹图谱,共标定了9个共有峰。36批紫苏叶与对照图谱相比较,相似度均大于0.900,聚类分析可将同一产地样品聚为一类;样品的9个共有峰变量投影重要性(VIP)值显示,其中7个成分可作为评价紫苏叶差异的指标。样品中野黄芩苷、迷迭香酸的含量测定方法学验证结果良好,36批紫苏叶中野黄芩苷、迷迭香酸质量分数分别为1.911%~4.968%、2.278%~10.257%,不同产地的样品含量差异较大。结论建立的指纹图谱和含量测定方法稳定性较高,分离度良好,含量测定结果结合指纹图谱化学模式识别分析结果表明不同产地的紫苏叶中主要成分差异较明显,可为紫苏叶的质量评价提供依据。

Objective To establish the fingerprint of Zisuye(Perillae Folium)from different regions and determine the content of scutellarin and rosmarinic acid,so as to provide reference for the quality control of Perillae Folium.Methods A total of 36 batches of Perillae Folium from nine producing areas were collected.HPLC was used to determine the contents of scutellarin and rosmarinic acid in Perillae Folium with acetonitrile-0.2%formic acid as the mobile phase.Cluster analysis,principal component analysis and partial least squares discriminant analysis were carried out by using Wekemo Bioincloud and SIMCA 14.1 tools.Results The fingerprint of Perillae Folium was established with scutellarin as the reference peak,and nine common peaks were calibrated.Compared with the control map,the similarity of 36 batches of Perillae Folium was more than 0.900.The samples from the same producing area could be clustered into one group by cluster analysis,and the VIP values of nine common peaks of the samples showed that seven components could be used as indexes to evaluate the difference of Perillae Folium.The content of scutellarin and rosmarinic acid in 36 batches of Perillae Folium was 1.911%—4.968%and 2.278%—10.257%,respectively.Conclusion The fingerprint and content determination method established in this study has high stability and good separation.The content determination results combined with fingerprint chemical pattern recognition analysis showed that the main components of Perillae Folium from different regions were significantly different,which could provide a basis for the quality evaluation of Perillae Folium.