三叶青藤叶中总黄酮、荭草苷和异荭草苷提取分离方法及HPLC检测条件的研究

Study on Extraction Methods and HPLC Detection Conditions of Flavone,Orientin and Isoorientin from Tetrastigma hemsleyanum

目的:建立三叶青藤叶中总黄酮、荭草苷及异荭草苷的提取分离和含量测定方法。方法:分别选取不同浓度的乙醇、丙二醇和丁二醇作为溶剂对三叶青藤叶中的有效成分进行提取,分别筛选出对总黄酮、荭草苷及异荭草苷提取率最高的溶剂,并采用高效液相色谱法(HPLC)进行探究。结果:选取荭草苷提取分离溶剂为30%丙二醇,异荭草苷的提取分离溶剂为40%乙醇。荭草苷的线性关系方程为Y=1.0524X-0.5086,在0.4~4μg/mL范围内的峰面积与进样浓度线性关系良好(R2=0.9997)。采用30%丙二醇提取三叶青藤叶中荭草苷的平均含量为3.58μg/g。异荭草苷的线性关系方程为Y=1.6650X-0.8264,在0.4~4μg/mL范围内的峰面积与进样浓度线性关系良好(R2=0.9996)。采用40%乙醇提取三叶青藤叶中异荭草苷的平均含量为1.47μg/g。结论:建立了三叶青藤叶总黄酮、荭草苷及异荭草苷的提取分离和含量测定方法。建立的高效液相色谱法方法简便、准确,重复性好,可应用于三叶青中这几种成分的含量测定。

Objective:To establish a method for the extraction,separation and determination of total flavone,orientin and isoorientin from Tetrastigma hemsleyanum leaves.Methods:Different concentrations of ethanol,propylene glycol and butanediol were selected as solvents to extract effective omponents from T.hemsleyanum leaves.The solvents with the highest extraction rates for total flavone,orientin and isoorientin were selected and investigated using high performance liquid chromatography(HPLC).Results:The solvent was 30%propylene glycol for orientin extraction and 40%ethanol for isoorientin extraction.The linear equation of orientin was Y=1.0524X-0.5086,and the peak area had a good linear relationship with injection concentration in the range of 0.4~4μg/mL(R2=0.9997).The average content of orientin in rhizome of Polygonum triphyllum was 3.58μg/g by 30%propylene glycol extraction.The linear equation of isoorientin was Y=1.6650X-0.8264,and the peak area had a good linear relationship with injection concentration in the range of 0.4~4μg/mL(R2=0.9996).The average content of isoorientin in T.hemsleyanum leaf was 1.47μg/g by 40%ethanol extraction.Conclusion:A method for the extraction,separation and determination of total flavone,orientin and isoorientin from T.hemsleyanum leaves was established.The established HPLC method is simple,accurate and reproducible,and can be applied to the determination of several components in T.hemsleyanum.