HPLC指纹图谱结合化学模式识别的不同采收期酸枣仁质量对比研究

Comparative Study on Quality of Ziziphi Spinosae Semen in Different Harvest Periods Based on HPLC Fingerprints Combined with Chemical Pattern Recognition

目的建立不同采收期酸枣仁HPLC指纹图谱分析方法,通过指纹图谱动态变化及化学模式识别方法比较不同采收期酸枣仁化学成分指纹峰的差异,寻找差异性标志物,为酸枣仁最佳采收期的确定提供依据。方法采用HPLC,Agilent EC-C18色谱柱(150 mm×4.6 mm,4μm),以乙腈-0.1%磷酸水为流动相,梯度洗脱,流速1.0 mL/min,检测波长312 nm。采集不同采收期酸枣仁药材指纹图谱,对不同采收期酸枣仁进行相似度评价,运用聚类分析、主成分分析及正交偏最小二乘-判别分析(OPLS-DA)对样品进行化学模式识别研究。结果建立了酸枣仁药材的HPLC指纹图谱,共确定了14个共有峰,指认了2个成分,分别为斯皮诺素(4号峰)、6’’’-阿魏酰斯皮诺素(9号峰);不同采收期的酸枣仁样品间相似度为0.906~1.000,不同的化学模式识别方法可对样品进行较好区分,OPLS-DA筛选出斯皮诺素等3个成分可能是影响酸枣仁质量的差异性标志物。结论通过HPLC指纹图谱和化学模式识别能够较好辨识影响酸枣仁质量的标志性成分,可为其最佳采收期的确定及质量控制提供依据。

Objective To establish the analytical method of HPLC fingerprints of Ziziphi Spinosae Semen in different harvest periods;To compare the differences in the fingerprint peaks of the chemical constituents of Ziziphi Spinosae Semen in different harvest periods through the dynamic changes of fingerprints and chemical pattern recognition methods;To look for differential markers;To provide a basis for the determination of the best harvest time of Ziziphi Spinosae Semen.Methods HPLC method was established on Agilent EC-C18 column(150 mm×4.6 mm,4μm)with acetonitrile-0.1%phosphoric acid solution as the mobile phase for gradient elution.The flow rate was 1.0 m L/min;chromatogram fingerprints was established at 312 nm.Fingerprints of Ziziphi Spinosae Semen in different harvest periods were collected to evaluate the similarity.Cluster analysis,principal component analysis and orthogonal partial least squares-discriminant analysis(OPLS-DA)were used to conduct chemical pattern recognition research on samples.Results The analytical method of HPLC fingerprints of Ziziphi Spinosae Semen were established.A total of 14 common peaks were identified,and 2 components were identified,namely spinosine(peak No.4)and 6’’’-feruloyl spinosine(peak No.9).The range of similarity of samples in different harvest periods was 0.906–1.000.The samples could be distinguished by different chemical pattern recognition methods.Spinosine and other three components may be differential markers that affect the quality of Ziziphi Spinosae Semen by OPLS-DA.Conclusion The iconic components that affect the quality of Ziziphi Spinosae Semen can be preferably identified by HPLC fingerprints and chemical pattern recognition methods.This research can provide a reference for the best harvest period and quality control of Ziziphi Spinosae Semen.