吴茱萸UPLC指纹图谱的建立及质量评价

Quality Evaluation of Evodiae Fructus by UPLC Fingerprint

目的:建立吴茱萸超高效液相色谱(UPLC)指纹图谱分析方法,对不同品种、不同产地吴茱萸药材的质量进行较全面的评价。方法:采用ACQUITY UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7μm),以乙腈-0.1%甲酸水为流动相进行梯度洗脱,检测波长300,215 nm,流速0.4 m L·min^(-1),柱温35℃。采用相似度评价、聚类分析及偏最小二乘法判别分析对37批样品的指纹图谱进行分析。结果:建立了吴茱萸药材的UPLC指纹图谱及共有模式,标定了34个共有峰,采用对照品比对,指认出17个共有峰。37批样品的相似度在0.703~0.973。不同来源的吴茱萸药材指纹图谱轮廓基本一致,化学成分的相对含量在较大范围内波动。聚类分析和偏最小二乘判别分析结果表明,化学成分的相对含量与药材品种、产地没有明显的相关性,而与药材的千粒重在一定程度上相关;吴茱萸次碱、吴茱萸碱、新绿原酸、绿原酸、吴茱萸卡品碱、柠檬苦素是差异较大的6个成分。结论:该方法简便、快速,可有效、全面地用于吴茱萸药材的质量评价;控制吴茱萸药材质量的关键是把握"近成熟"这一采收时间环节。

Objective： To establish the ultra performance liquid chromatography （UPLC） fingerprint of Evodiae Fructus, and provide a comprehensive evaluation for their quality from different species and regions. Method： The UPLC method was performed on ACQUITY UPLC BEH C18 （2.1 mm × 100 mm, 1.7 μm） , with acetonitrile-0.1% formic acid aqueous solution as mobile phase for gradient elution at a flow rate of 0.4 mL·min- The detection wavelength was set at 300 and 215 nm, the column temperature was set at 35 ℃. Similarity evaluation, hierarchical clustering analysis and partial least squares-discriminate analysis were applied to evaluatethe fingerprints of 37 batches of Evodiae Fructus. Result： The UPLC fingerprint and common mode were established with 34 common peaks, and 17 of them were identified based on the reference substances. The similarities of 37 samples were 0. 703-0. 973. And fingerprint profiles of various samples from different regions were basically similar, but the relative contents of components varied greatly. Based on the results of hierarchical clustering analysis and partial least squares-discriminate analysis, the relative contents of components were almost unrelated to the species and regions, but related to 1 000 grain-weight of the fruits to a certain extent. Rutaecarpine, evodiamine, neochlorogenic acid, chlorogenic acid, evocarpine and limonin turned out to be the top 6 differential compounds. Conclusion： This simple and rapid method could be effectively used for the comprehensive quality evaluation of Evodiae Fructus. It is the most important to confirm that near mature collection time is the key to guarantee the quality of crude drug of Evodiae Fructus.