HPLC同时测定红芪中8个活性成分的含量及聚类分析

Determination of eight active components in Radix Hedysari by HPLC and its cluster analysis

建立了HPLC法测定红芪中腺苷、γ-氨基丁酸(GABA)和6个黄酮类化合物(毛蕊异黄酮苷、芒柄花苷、毛蕊异黄酮、异甘草素、芒柄花素和美迪紫檀素)的含量。样品用甲醇回流提取4 h,采用HPLC-DAD(二极管阵列)和HPLC-ELSD(蒸发光散射)系统,色谱柱为Diamonsil C_(18)柱(250 mm×4.6 mm,5μm),流动相为乙腈-水,柱温40℃,流速1.0 m L·min^(-1),漂移管温度110.5℃,载气流速3.1 L·min^(-1)。结果 8个成分的线性关系良好(r^2=0.992 8~1.000 0);精密度、稳定性和重复性的RSD均小于2%;加样回收率为96.78%~103.45%,RSD为0.29%~1.61%。测定24批不同产地红芪的指标成分含量,并以其作为变量因子进行聚类分析,结果基本按地域性聚成2类并与主成分分析结果一致。该方法准确、灵敏、简便,为多指标控制红芪药材质量提供了实验依据;将聚类分析用于红芪质量控制具有一定的可靠性和客观性。

An HPLC method was established for the determination of adenosine,γ-aminobutyric acid（GABA） and six flavonoids（calycosin-7-glucoside,ononin,calycosin,isoliquiritigenin,formononetin and medicarpin） in Radix Hedysari.The samples were extracted with methanol by refluxing for 4 h.The HPLC-DAD was performed on a Diamonsil C18 column（250 mm × 4.6 mm,5 μm） with acetonitrile-water as the mobile phase.The column temperature was at 40 ℃ and the flow rate was 1.0 mL·min^-1,while the temperature of drift tube was 110.5 ℃ and the nebulizing gas flow was 3.1 L·min^-1 for the ELSD system.The results showed all the eight components had good linear relationships（r-2 =0.992 8-1.000 0） in the range of the test concentration.The RSD of precision,stability and repeatability were less than 2%.The average recovery rates were 96.78%-103.45%,and RSD were 0.29%-1.61%.The index component contents of Radix Hedysari form 24 different origins were determined and used as variable factors in clustering analysis.The results were classified into 2 groups basically in accordance with the regional cluster.And the consequence was in consistent with the results of principal component analysis.This HPLC method is simple,shows good sensitive and accurate,and provides the experimental basis for multi-index control of Radix Hedysari.Clustering analysis for Radix Hedysari quality control has a certain reliability and objectivity.