HPLC同时测定毛蕊花糖苷、咖啡酸和羟基酪醇

Simultaneous determination of acteoside,caffeic acid and hydroxytyrosol by HPLC

根据毛蕊花糖苷、羟基酪醇、咖啡酸的紫外吸收性质,建立了同时检测3种物质的高效液相色谱法。采用Waters e2695型高效液相色谱仪,色谱柱为Waters Symmetry C18柱(5μm,250 mm×4.6 mm);流动相为甲醇(A)和体积分数0.1%甲酸水溶液(B),比例为40:60(V/V);流速0.7 m L/min;2998PDA二极管阵列检测器,检测波长280 nm,柱温35℃;进样量10μL。毛蕊花糖苷标准曲线的线性回归方程为y=4.474ρ+5.224,R2=0.9999;加样回收率为100.2%。咖啡酸标准曲线的线性回归方程为y=20.687ρ-3.392,R2=0.9995;加样回收率为100.0%;羟基酪醇标准曲线的线性回归方程为y=4.716ρ-17.901,R2=0.9999;加样回收率为102.0%;毛蕊花苷的检测限为5 ng,咖啡酸的检测限为0.8 ng,羟基酪醇的检测限为0.25 ng。

According to the ultraviolet absorption properties of acteoside, caffeic acid and hydroxytyrosol, a simultaneous determination method for the three substances by HPLC was established. The HPLC chromatographic samples were separated on a Waters Symmetry C18 column （5 μm, 250 mm×4. 6mm） and detected by 2998 photo-diode-array detector （PAD）. The mobile phase was methanol - 0. 1% aqueous formic acid （40：60, V/V）. The Analysis was performed at 30℃ with a flow rate of 0. 7 mL/min and detection at 280 nm. The sample volume was 10 μL. The standard curve equation of aeteoside was y = 4. 474p ＋ 5.224, R^2 =0. 9999 and the average recovery was 100.2%. The standard curve equation of caffeic acid was y =20. 687p - 3.392, R^2 = 0. 9995 with the average recovery of 99. 97%. The standard curve equation of hydroxytyrosol was y =4. 716p - 17. 901, R^2 = 0. 9999 and the average recovery was 102. 0%. The detection limits of acteoside, caffeic acid and hydroxytyrosol were 5 ng, 0. 8 ng and 0. 25 ng, respectively. The results indicated that this methodology is simple, accurate and reproducible.