车前子的HPLC特征图谱研究

The Study of HPLC Specific Chromatogram for Plantaginis Semen

目的建立车前子的HPLC特征图谱。方法采用HPLC法测定,使用Agilent Eclipse XDB-C18色谱柱(250mm×4.6mm,5μm),以甲醇-0.1%甲酸水溶液为流动相,梯度洗脱,流速1.0 m L·min-1;检测波长250nm,柱温为30℃。对10个批次车前子检测,采用国家药典委员会中药色谱指纹图谱相似度评价系统软件(2004A版)进行数据处理分析。并用HPLC-MS技术对共有峰进行了初步归属并比较了车前子与平车前子HPLC图谱的差异。结果建立了以3个共有峰为指标成分的车前子药材的HPLC特征图谱,通过MS定性分析结合对照品比对,确定共有峰分别为京尼平苷酸、毛蕊花糖苷和异毛蕊花糖苷,车前子与平车前子的HPLC图谱差异明显。结论该方法经过方法学验证,可用于车前子的质量控制。

OBJECTIVE To establish HPLC specific chromatogram of Plantaginis semen. METHODS The a- nalysis was performed on an Agilent Eclipse XDB-Cls column （250mm ×4. 6mm ,5 μm）with gradient mobile phase of methanol-0. 1% formic acid aqueous solution at a flow rate of 1.0mL · min-1. The detection wavelength was set at 250nm,and the column temperature was 30℃. The data were analyzed by the National Pharmacopoeia Committee of Chinese Pharmacopoeia Chromatographic Fingerprint Similarity Evaluation System software （2004A Version）. And part of the common peaks were preliminarily assigned by HPLC-MS. The differences of HPLC chromatogram of two species Plantaginis semen were compared. RESULTS The HPLC specific chromatogram of the Plantaginis semen samples were established, and three common peaks were identified as Geniposidic acid, Verbaseoside and Isoacteoside by qualitative analysis. The HPLC chromatogram were obvious differences between two species Plantaginis semen. CONCLUSION The developed method conforms to the requirements of methodology validation, which can be used for the quality control of Plantaginis semen.