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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 a HPLC fingerprinting and cluster analysis method for white back leaf medicinal materials and provide a scientific basis for the scientific evaluation and effective control of white back leaf medicinal materials. Methods: Fingerprints of white dorsal leaf from 12 different producing areas were established by HPLC. Phenomenex-C18 column (4.6 mm × 250 mm, 5 μm) was used. The mobile phase consisted of methanol-0.1% glacial acetic acid Off, flow rate 1.0 m L · min-1, column temperature 25 ℃, detection wavelength 336 nm. Similarity analysis software was used to carry out similarity analysis, and SPSS 19.0 software was used to carry out cluster analysis. The relative peak area of common peaks was used as index to standardize and analyze PCA. Results: The common pattern of HPLC fingerprints of white backs and backs was established, and the similarity of white back leaf medicinal materials from different areas were compared. The fingerprints were also analyzed by cluster analysis and principal component analysis. 22 common peaks were calibrated and 5 common peaks were identified. According to the similarity calculation, the overall similarity was 0.554 ~ 0.981. The cluster analysis divides the samples 2, 3, 4, 5, 6, 8, 9, 10 and 12 into two categories when the distance scale is 2, the comprehensive principal component score (F value) The samples of 7, 1, 9, 12, 6, 5, 10, 4, 3, 2 and 8 samples were classified according to the two methods. Conclusion: The method is simple, accurate, sensitive and reproducible. It provides a scientific basis for the research of white back leaf medicinal materials and the establishment of quality control standards.