超高效液相色谱法测定不同产地槐花中4种黄酮含量

Determination of 4 flavonoids in Sophora japonica from different habitats by ultra performance liquid chromatography

目的建立超高效液相色谱法(ultra performance liquid chromatography,UPLC)检测槐花中4种黄酮类成分含量的分析方法,并比较不同产地槐花中4种黄酮类成分的含量差异及聚类分析。方法色谱柱为ACQUITY UPLC BEH C18(2.1 mm×50 mm,1.7μm);流动相A为甲醇溶液,B为0.1%乙酸水溶液,梯度洗脱。柱温为30℃,检测波长为360 nm;通过TB tools热图绘制选项以及PCA主成分分析图绘制功能进行各产地样品的聚类分析。结果 4种黄酮对照品在相应浓度范围内与峰面积均呈现良好线性关系;回收率均在95%~105%之间;不同产地槐花中的4种黄酮类成分含量差异较大,基于4种黄酮类成分含量对不同产地的槐花的PCA主成分分析及聚类热图分析结果表明,槐花具有明显的产地聚类特性。结论该方法操作简单,结果稳定准确,专属性强,并且分析时间短,能够降低分析成本,适合作为槐花的含量测定方法。不同产地槐花质量差异明显,可通过优选产地从而保证其来源的质量稳定性以及用药安全性。

Objective To establish a method for the determination of 4 flavonoids in Sophora japonica by ultra performance liquid chromatography(UPLC),and to compare the content differences and cluster analysis of the 4 kinds of flavonoids in Sophora japonica from different regions.Methods The column was ACQUITY UPLC BEH C18 column(2.1 mm×50 mm,1.7μm).Mobile phase A was methanol solution,and B was 0.1% acetic acid aqueous solution,with gradient elution.The column temperature was 30℃,and the detection wavelength was 360 nm.The cluster analysis of samples from different origins was carried out through the TB tools heat map drawing option and the PCA principal component analysis map drawing function.Results Four kinds of flavonoid reference substances showed a good linear relationship with the peak area in the corresponding concentration range.The recovery rates were all between 95% and 105%.The content of 4 kinds of flavonoid components in Sophora japonica from different origins varied greatly,based on 4 kinds of flavonoids in PCA principal component analysis and clustering heat map analysis of locust flower from different origins showed that locust flowers had obvious clustering characteristics of origin.Conclusion This method is simple in operation,stable and accurate in results,strong in specificity,and short in analysis time,which can reduce the analysis cost,and is suitable as a content determination method for Sophora japonica.The quality of Sophora japonica from different producing areas is obviously different,so the quality stability and medication safety of Sophora japonica can be ensured by optimizing the producing areas.