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 = 1.052 4X-0.508 6,在0.4~4 μg/mL范围内的峰面积与进样浓度线性关系良好(R2 = 0.999 7).采用30%丙二醇提取三叶青藤叶中荭草苷的平均含量为3.58 μg/g.异荭草苷的线性关系方程为Y = 1.665 0X-0.826 4,在0.4~4 μg/mL范围内的峰面积与进样浓度线性关系良好(R2 = 0.999 6).采用40%乙醇提取三叶青藤叶中异荭草苷的平均含量为1.47 μg/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 fla-vone,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 isoorien-tin 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 = 1.052 4X-0.508 6,and the peak area had a good linear relationship with injection concentration in the range of 0.4~4 μg/mL(R2 = 0.999 7).The average content of orientin in rhizome of Polygonum triphyllum was 3.58 μg/g by 30%propylene glycol extraction.The linear equa-tion of isoorientin was Y= 1.665 0X-0.826 4,and the peak area had a good linear relationship with injec-tion concentration in the range of 0.4~4 μg/mL(R2 = 0.999 6).The average content of isoorientin in T.hemsleyanum leaf was 1.47 μg/g by 40%ethanol extraction.Conclusion:A method for the extraction,sep-aration and determination of total flavone,orientin and isoorientin from T.hemsleyanum leaves was estab-lished.The established HPLC method is simple,accurate and reproducible,and can be applied to the de-termination of several components in T.hemsleyanum.