天然酚酸类化合物的反相高效液相色谱分析

Studies on the Separation and Determination of Natural Phenolic Acids by Reversed-Phase High Performance Liquid Chromatography

采用反相高效液相色谱法梯度洗脱 ,对 12种天然酚酸进行定性定量分析研究。通过色谱条件的优化 ,确定了最佳的分离条件 ;同时探讨了流动相的酸度对分离结果的影响 ,以及酚酸结构与保留行为之间的关系。实验中以确定的最佳分离条件对 12种天然酚酸进行了定量分析 ,测定了各个化合物的回归方程、相关系数、线性范围、检测限和相对标准偏差。实验结果显示 ,各个化合物回归方程的相关系数r为 0 9980～ 0 9999,检测限为 0 96ng～ 4 10ng ,相对标准偏差RSD≤ 2 6 4 % ,满足定量分析要求。

A new method using gradient elution of reversed phase high performance liquid chromatography was set up for the separation and determination of 12 natural phenolic acids simultaneously. The separation conditions were studied with a Zorbax ODS (5 μm) column, and optimized conditions were suggested with the mobile phase gradient elution of CH 3CN H 2O (with 0 5% HAc) (5∶95→ 35∶65, V/V , in 40 min) at room temperature, flow rate at 1 0 mL/min and detection wavelength at 280 nm. And the effects of acidity on separations and the relationships between the phenolic acids structures and their retention behaviors were discussed. Qualitative and quantitative analysis of the 12 compounds were performed. The regression equation, correlation coefficient, linear range, detection limit and relative standard deviation (RSD) of each compound were obtained under the optimized conditions. The correlation coefficients of the 12 compounds were between 0 998?0 and 0 999?9 . The detection limits ranged from 0 96 ng to 4 10 ng and RSDs were ≤2 64%. These results demonstrate that the optimized determination method obtained is successful for the separation and determination of the 12 phenolic acids analysed.