基于UPLC指纹图谱结合薄层色谱的白及药材质量标准探讨

Study on the Quality Standard of <i>Bletilla striata</i> (Thunb.) Reichb.f. Based on UPLC Fin-gerprint and Thin Layer Chromatography

采集60批不同产地白及进行薄层色谱及超高效液相色谱测定,为白及质量评价体系改进提供实验依据。采用薄层层析法(TLC)鉴别是否为白及正品;采用Waters ACQUITY UPLC BEH C18色谱柱(2.1 mm &#215;100 mm, 1.7 μm),以乙腈(A)和0.1%甲酸水(B)为流动相进行梯度洗脱,流速0.208 mL?min?1,柱温35℃,检测波长270 nm。以贝母兰宁为参照,绘制61批药材样品的UPLC图谱,参照白及对照药材进行相似度评价及聚类分析。60份样品中有15个产地的样品经TLC鉴定合格;建立了白及药材UPLC指纹图谱,确定8个共有峰,不同供试白及指纹图谱与特征图谱相似度在0.623~0.994之间,聚类分析的距离为0.25时可聚为4类。本研究发现以薄层色谱的特定斑点作为白及鉴别依据不够充分;建立了白及UPLC指纹图谱,为白及药材质量控制提供参考。

The samples from different producing areas were collected and determined by thin layer chromatography (TLC) and ultra-high performance liquid chromatography (UPLC), which provided theoretical and experimental basis for the improvement of quality evaluation system. Bletilla striata (Thunb.) Reichb.f. authentic were identified by TLC according to Chinese Pharmacopoeia. UPLC analysis was performed on Waters ACQUITY UPLC BEH C18 column (2.1 mm &#215;100 mm, 1.7 μm), the column temperature was 35?C, and the mobile phase was acetonitrile (A) and 0.1% formic acid water (B). The flow rate was 0.208 mL?min?1 with detection wavelength of 270 nm. Taking coelonin as a reference, the UPLC maps of 61 batches of medicinal materials were drawn, and similarity evaluation and cluster analysis were carried out with reference to B. striata. Among the 60 samples, 15 samples were qualified by TLC. The UPLC fingerprint of B. striata was established, and eight common peaks were determined. The similarity was between 0.623~0.994. The results of cluster analysis showed that it could be divided into four groups when the similarity coefficient was 0.25. This study found that the use of specific spots in thin layer chromatography as the basis for B. striata identification was insufficient;B. striata UPLC fingerprints were established to provide a reference for the quality control of B. striata medicinal materials.