目前，万古霉素色谱分析方法主要为反相色谱法，该法分离万古霉素及其杂质时，存在极性选择性不足以及所使用的流动相体系与质谱兼容性差等问题。基于亲水作用色谱（ HILIC）对糖肽类物质的色谱保留和极性选择性，本文选取万古霉素及其常见杂质为对象，考察了HILIC固定相、流动相组成比例、缓冲盐添加剂浓度和pH值等色谱条件，对万古霉素及其主要杂质进行了HILIC分离方法研究。确立了以Click XIon色谱柱为固定相，以甲酸铵为流动相添加剂的亲水作用色谱条件，实现了万古霉素及主要杂质的分离，为万古霉素类化合物的分离提供了新的途径。
The current reversed?phase liquid chromatography as the main analytical method for vancomycin is inappropriate due to its poor polarity selectivity and mass spectrometry compati?bility. Hydrophilic interaction chromatography ( HILIC) method has been verified to provide the glycopeptide substances with good retention and polarity selectivity, therefore, an HILIC meth?od was developed for the analysis of vancomycin and its related impurities including norvanco?mycin, desvancosaminyl vancomycin, dedichloro vancomycin and crystalline degradation prod?uct ( CDP?1) . In this work, a systematic evaluation of retention behavior of vancomycin and its related impurities in hydrophilic interaction chromatography was performed and the influences of stationary phase, acetonitrile content, buffer salt and pH of mobile phase were studied. According to the results, Click XIon column was selected as stationary phase and ammonium formate was additive in the mobile phase. Vancomycin and its related impurities were totally separated under the optimization conditions. It is concluded that hydrophilic interaction chroma?tography may be a new strategy for the separation of vancomycin and its structural analogues.
参考文献
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