高效液相色谱法中不同pH值对黄酮及有机酸分离效果的探讨

Discussion on separate effect of flavone and organic acid at different pH value in High Performance Liquid Chromatography

目的探讨不同pH值在高效液相色谱法中对黄酮苷有机酸类化合物的分离效果的影响。方法色谱柱为AgilentHC-C18,流动相：乙腈（A）-（0.05mol/L）KH2PO4-H3PO4缓冲溶液pH=3.5、3.1、3.0、2.5（B）,线性洗脱梯度,洗脱程序如下：0～65min乙腈10%→28%,65~85min乙腈28%→52%。检测波长：220nm;流速：1mL/min。结果流动相pH值的不同对黄酮及有机酸色谱峰的出峰时间、峰形、分离度均有较大的影响。pH=3.0色谱峰形比较好,黄酮及有机酸达到较好的分离效果。结论本研究体现了pH值调节在高效液相色谱中的作用,为有效分离黄酮及有机酸提供依据。

Objective The aim was to discuss the separate effect of flavone and organic acid at different pH value in HPLC. Methods The separation was performed on Agilent HC-C18, The mobile phase consisted of acetonitrile （A） with a-（0.05mol/L） KH2PO4-H3PO4 buffer solution （B）. The pH value was adjusted to 3.5,3.1,3.0,2.5. The detecting wavelength was set at 220 nm and the flow rate was 1.0ml/min. Results The pH valu of mobile phase made great influence on the time of occurring peaks, the shape of peaks and separation. At a pH value of 3.0,both Flavone and Organic Acid separated well and their peaks were in good shape. Conclusion This method shows the effect on the pH value adjustment. It provided bases for separation Flavone and Organic Acid.