单糖柱前PMP衍生HPLC测定方法探讨

Research on the Method of PMP Pre-column Derivation of Monosaccharides by HPLC

从PMP萃取酸度、萃取次数探讨了柱前PMP衍生方法对单糖-PMP衍生物的影响,并从流动相比例、缓冲液pH值、缓冲盐浓度三个方面对单糖柱前衍生HPLC的分离效果进行了优化研究.结果表明:单糖进行PMP衍生后,用HCl中和至pH为7.0,能最大限度避免单糖-PMP衍生产物的损失;使用三氯甲烷萃取PMP三次时,能在除去PMP的情况下避免单糖-PMP衍生产物的损失;降低流动相中有机相的比例或提高缓冲液的pH值,有利于色谱峰分离度的提高;提高缓冲盐浓度,有利于保持色谱峰的对称性.

This article discussed the impact of PMP pre-column derivation on monosaccharide derivatives from extraction pH and extraction times. Moreover, the influence of mobile phase on the HPLC chromato- graphic peaks also have been studied from the ratio of mobile phase, the pH of buffer and the concentra- tion of buffer. The results indicated ：Neutralized the derivation liquid with HC1 to pH7.0 could avoid the loss of derivatives products to the utmost.Extracting three times by chloroform could remove almost PMP and avoid the loss of derivatives.Reducing the ratio of organic phase or the pH of buffer could improve the seperation of peaks;the asymmetry factor of peaks could be maintained in the optimum range by increasing the concentration of buffer.