HPLC检测三叶青块根中白藜芦醇和白藜芦醇苷含量的研究

Determination of Resveratrol and Polydatin in the Ground and Underground Parts of Tetrastigma hemsleyanum by HPLC

目的:建立三叶青地下部分白藜芦醇、白藜芦醇苷以及总黄酮的提取分离和含量测定方法。方法:白藜芦醇提取分离选取溶剂为70%乙醇,白藜芦醇苷的提取分离选取溶剂为40%乙醇。含量测定采用高效液相色谱法,色谱柱选择Agilent ZORBAX SB-C18色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇-水梯度洗脱,流速为0.8 mL/min,检测波长为304 nm,柱温25℃。结果:白藜芦醇的线性关系方程为Y=113.836X+0.9115,在0.02~2μg/mL范围内的峰面积与进样浓度线性关系良好(R^(2)=0.9976)。采用70%乙醇提取三叶青根茎中白藜芦醇的平均含量为0.59μg/g。白藜芦醇苷的线性关系方程为Y=52.515X-0.4977,在0.02~2μg/mL范围内的峰面积与进样浓度线性关系良好(R^(2)=0.9944)。采用40%乙醇提取三叶青根茎中白藜芦醇的平均含量为1.76μg/g。结论:建立了三叶青地下部分白藜芦醇、白藜芦醇苷以及总黄酮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,可应用于三叶青中单体有效成分的含量测定。

Objective:To establish a method for the extraction and content determination of resveratrol,polydatin,and total flavonoids in subterranean part of Tetrastigma hemsleyanum.Methods:70%ethanol was selected as the solvent for extraction and separation of resveratrol and 40%ethanol as the solvent for extraction and separation of resveratrol glycosides.HPLC was used to determine the content.Agilent ZORBAX SB-C18 column(250 mm×4.6 mm,5μm)was selected.The mobile phase was methanolwater gradient elution,the flow rate was 0.8 mL/min,the detection wavelength was 304 nm,and the column temperature was 25℃.Results:The linear equation of resveratrol was Y=113.836X+0.9115,and the linear relationship between the peak area and the injection concentration was good in the range of 0.02~2μg/mL(R^(2)=0.9976).The average content of resveratrol in T.hemsleyanum extracted by 70%ethanol was 0.59μg/g.The linear equation of resveratrol glycosides was Y=52.515X-0.4977,and there was a good linear relationship between the peak area and injection concentration in the range of 0.02~2μg/mL(R^(2)=0.9944).The average content of resveratrol was 1.76μg/g in T.hemsleyanum extracted by 40%ethanol.Conclusion:A method has been established for the extraction and content determination of resveratrol,polydatin,and total flavonoids in the subterranean part of T.hemsleyanum.The established HPLC method is simple,accurate and reproducible,and can be used for the determination of monomeric active ingredients in T.hemsleyanum.