白背叶HPLC特征指纹图谱及模式识别

HPLC Fingerprint and Pattern Recognition of Liposoluble Compontents in Mallotus apelta Leaves

目的:建立白背叶药材的HPLC指纹图谱与系统聚类分析方法,为科学评价和有效控制白背叶药材的质量提供科学依据。方法:采用高效液相色谱法建立12个不同产地白背叶的指纹图谱,采用phenomenex-C18色谱柱(4.6 mm×250 mm,5μm);以甲醇-0.1%冰乙酸水为流动相,梯度洗脱,流速1.0 m L·min-1,柱温25℃,检测波长336 nm。并使用相似度评价软件进行相似度分析,用SPSS 19.0软件进行聚类分析,以共有峰的相对峰面积为指标,进行标准化处理和主成分分析。结果:建立了白背叶HPLC指纹图谱共有模式,对不同产地白背叶药材进行了相似度比较,并进行了指纹图谱系统聚类分析和主成分分析。标定了22个共有峰,指认了其中5个共有峰,经相似度计算,整体相似度为0.554~0.981。聚类分析在距离标尺为2时将样品2,3,4,5,6,8,9,10,12分为一大类;综合主成分得分(F值)从高到低依次为11,7,1,9,12,6,5,10,4,3,2,8号样品,F在0.0^-1.7共2种方法的分类结果一致,可以相互验证。结论:该方法简便准确、灵敏度高、重复性好,为白背叶药材的研究以及制定质量控制标准提供科学依据。

Objective： To establish the HLPC fingerprint and cluster analysis methods of Mallotus apelta leaves,in order to provide a reasonable basis for scientific evaluation and quality control of M. apelta leaves.Method： Chromatographic fingerprints of M. apelta leaves from 12 different origins were established by HPLC. A phenomenex-C18column（ 4. 6 mm × 250 mm,5 μm） was adopted and eluted with the mobile phase of methanol and 0. 1% acetic acid in a gradient mode. The flow rate was 1. 0 m L·min^（-1）,the column temperature was 25 ℃,and the detection wavelength 336 nm. The similarity was analyzed by ‘Similarity Evaluation System for Chromatographic Fingerprint of TCM＇,and SPSS 19. 0 was applied in a cluster analysis. The relative peak area of common peaks was taken as the index for standardized treatment and principal component analysis. Result： The mutual mode of HPLC fingerprints was set up,and the similarity between crude drugs of different habitats were compared. SPSS was used for fingerprint system cluster analysis and principal component analysis. Totally 22 common peaks were identified as markers,and 5 common components were identified in HPLC fingerprint of M.apelta leaves. The overall similarity was calculated to be 0. 554-0. 981. When the distance scale of cluster analysis was 2,the samples 2,3,4,5,6,8,9,10,12 were classified into one category. The order for comprehensive principal component scores（ F） from high to low were samples 11,7,1,9,12,6,5,10,4,3,2,8. Two results of the two methods can be mutually verified when F was between 0. 0 and-1. 7. Conclusion： The method is accurate,simple,stable and highly repeatable,and can provide scientific basis for studying crude drug of M.apelta leaves and making quality control standards.