水黄皮HPLC指纹图谱的优化及建立

目的通过筛选试样处理方法和优化水黄皮高效液相色谱分析方法,建立水黄皮的指纹图谱。方法实验比较了三种不同的提取方法和提取时间,考察了不同型号色谱柱的分离效果和多种流动相系统的梯度洗脱能力,优化了检测波长、柱温和流速等分析条件。色谱柱为Agilent ZORBAX SB C18(250 mm×4.6 mm,3.5μm)柱,流动相为乙腈(A)-0.5%H3PO4(B)溶液,梯度洗脱;检测波长为303 nm;柱温为35℃;流速为1.0 m L/min。采用中药色谱指纹图谱相似度评价系统软件(2004 A版)进行指纹图谱分析。结果通过优化检测波长、流动相、洗脱程序等建立了水黄皮HPLC指纹图谱及共有模式,标定9个共有峰。9批样品的相似度为0.828~0.983。不同来源的水黄皮指纹图谱基本一致,所含化学成分的种类和含量较稳定。结论该方法相对于单一成分的质量控制模式,能够直观、全面地从整体上反映水黄皮药材的质量,准确可靠、重复性好,为水黄皮的鉴定和质量控制提供了参考依据。

Optimization of conditions and establishment of HPLC fingerprint of Pongamia pinnata

Objective To establish the fingerprint of Pongamia pinnata by screening the samples and optimizing the high performance liquid chromatography (HPLC) analysis method.Methods Three different extraction methods and extraction time were compared.The separation effect of different types of columns and the gradient elution ability of various mobile phase systems were investigated,and the analysis conditions such as detection wavelength,column temperature and flow rate were optimized.The HPLC method was performed with Agilent ZORBAX SB C18 column (250 mm×4.6 mm,3.5 μm),and the mobile phase was acetonitrile (A)-0.5％ H3PO4 (B) (gradient elution) with a flow rate of 1.0 mI/min.The detection wavelength was 303 nm,and the column temperature was 35 ℃.The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (2004 A edition) was used to analyze the results.Results The HPLC fingerprint of P.pinnata and its common mode were established with the optimization of the detection wavelengths,mobile phase and elution conditions.Nine common peaks were characterized.The similarities of nine batches were 0.828-0.983.The fingerprint profiles of various samples from different regions were basically identical,with the relatively stable type and content of the chemical compositions.Conclusion Compared with the single component quality control mode,this method can reflect the quality of P.pinnata in an intuitive and comprehensive way,and it is accurate,reliable and reproducible,which provides the reference for the identification and quality control of P.pinnata.