Development and validation of an improved method for the analysis of vancomycin by liquid chromatography selectivity of reversed-phase columns towards vancomycin components.
Development and validation of an improved method for the analysis of vancomycin by liquid chromatography selectivity of reversed-phase columns towards vancomycin components.
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DOI：
10.1016/s0021-9673(03)00535-1
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发表时间：
2003-05
期刊：
影响因子：
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通讯作者：
J. Diana;D. Visky;E. Roets;J. Hoogmartens
中科院分区：
文献类型：
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作者：
J. Diana;D. Visky;E. Roets;J. Hoogmartens
官方各论中规定的万古霉素纯度控制的现行方法不适当，因为几种组分未相互分离，其他组分与主要组分万古霉素B共洗脱。该方法使用pH 3.2的ODS柱。在这项研究中，引入了几个变化，以改善分离。在低pH下的分离方法的优化表明，pH 1.7是最佳的，并且使用二氧六环作为有机改性剂显著改善了分离。这些条件用于测试一组40多个反相柱对万古霉素组分的选择性。通过主成分分析选择最合适的色谱柱。这些色谱柱中的大多数不允许将二氯万古霉素与单氯万古霉素1分离。发现中性至弱碱性移动的相允许更好的分离。通过实验设计对分离方法进行了进一步优化，并进行了耐用性研究。该优化表明pH 7.7是最佳的，并且梯度洗脱也用于实现完全分析。最终方法使用Kromasil色谱柱，移动的相包含二氧六环、水和甲酸铵溶液（pH 7.7）。首次从主要成分万古霉素B中分离出单氯万古霉素2和一些未知杂质。该方法具有良好的重复性、线性和灵敏度。
The current method prescribed in official monographs for the purity control of vancomycin is inappropriate in that several components are not separated from each other and other components are coeluted with the main component vancomycin B. The method uses an ODS column at pH 3.2. In this study, several changes were introduced in order to improve the separation. The optimization of the separation method at low pH indicated that pH 1.7 was optimum and that the use of dioxane as organic modifier drastically improved the separation. These conditions were used to test a set of more than 40 reversed-phase columns for their selectivity towards vancomycin components. The selection of the most suitable columns was performed by means of principal component analysis. Most of these columns did not allow the separation of didechlorovancomycin from monodechlorovancomycin 1. It was found that neutral to slightly alkaline mobile phases allowed better separation. Further optimization of the separation method and a robustness study were performed by means of experimental design. This optimization indicated that pH 7.7 was optimum and gradient elution was also used to effect complete analysis. The final method uses a Kromasil column and the mobile phase comprises dioxane, water and ammonium formate solution pH 7.7. The separation of monodechlorovancomycin 2 and of some unknown impurities from the main component vancomycin B is described for the first time. The method shows good repeatability, linearity and sensitivity.