HPLC结合化学计量学方法用于不同生长年限甘草药材黄酮类成分特征图谱研究

Specific Chromatograms of Glycyrrhiza uralensis Fisch.Flavonoids in Different Growth Years by HPLC Coupled with Chemometric Analysis

目的建立甘草药材黄酮类成分高效液相色谱(HPLC)特征图谱,结合化学计量学方法对不同生长年限甘草药材进行特征图谱分析,研究不同生长年限甘草药材之间的差异,为其质量控制提供科学的评价方法。方法采用Shiseido Capcell Pak C18色谱柱(4.6 mm×250 mm,5μm);流动相为乙腈(A)-0.1%甲酸水(B),梯度洗脱;流速1.0 mL·min-1;检测波长280 nm;柱温:40℃。通过对照品比对,对建立的指纹图谱主要特征峰进行确认,采用Chem Pattern化学计量学软件对结果进行处理,运用相似度分析、主成分分析和聚类分析方法深入研究影响不同生长年限甘草黄酮类化学成分的关键指标。结果4年生甘草药材质量最佳。甘草苷、芹糖甘草苷、甘草素、异甘草苷的差异显著,可作为区分不同生长年限甘草的关键性指标成分。结论通过化学计量学分析方法,建立的甘草药材黄酮类成分特征图谱可为甘草药材规范化栽培和标准提高提供参考。

OBJECTIVE To establish the HPLC specific chromatogram of licorice flavonoids based on chemometric analysis method,and search for differentiated components of Glycyrrhiza uralensis Fisch.cultivated for different years.METHODS The analysis was performed on a Shiseido Capcell Pak C18 column(4.6 mm×250 mm,5μm)by gradient elution with acetonitrile-0.1%formic acid solution as mobile phase at a flow rate of 1.0 mL·min-1.The column temperature was maintained at 40℃and the detection wavelength was set at 280 nm.The HPLC specific chromatograms of Glycyrrhiza uralensis Fisch.cultivated for different years were further evaluated by chemometric methods including similarity analysis(SA),hierarchical clustering analysis(HCA)and principal component analysis(PCA).RESULTS Four-years-old licorice was confirmed as the best one.Liquiritin,liquiritin apioside,liquiritigenin and isoliquiritin had significant differences,which can be used as key indicators to distinguish Glycyrrhiza uralensis Fisch.cultivated for different years.CONCLUSION This method can provide reference for standardized cultivation and quality standard improvement of licorice.