Department of Inorganic Chemistry, Medical University of Lublin, Chodźki 4a Street, 20-093 Lublin, Poland.
J Chromatogr A. 2013 Oct 11;1311:48-54. doi: 10.1016/j.chroma.2013.06.036. Epub 2013 Jul 1.
A HPLC procedure on strong cation exchange column (SCX) has been developed for the analysis of selected alkaloids from different chemical groups. The retention, separation selectivity, symmetry of peaks and system efficiency were examined in different eluent systems containing different types or concentrations of buffers at various pH and the addition of organic modifiers: methanol (MeOH), acetonitrile (CH3CN), tetrahydrofuran (THF) or dioxane (Dx). The retention factors as the function of the concentration of buffers, the mobile phase pH and the percentage of modifier in the eluents were investigated. More symmetrical peaks and the highest theoretical plate number were obtained in eluents containing acetonitrile or tetrahydrofuran. In most cases, the increase of buffer concentration caused the decrease of alkaloids' retention, the improvement of peaks' symmetry and the increase of theoretical plate number. The improved peak symmetry and the efficiency of system for most investigated alkaloids were observed in the systems containing buffers at strongly acidic pH. The obtained results also reveal a large influence of salt cation used for buffer preparation. The results obtained on SCX column were compared with those obtained on a C18 column. The most efficient and selective systems were used for the separation of alkaloid standard mixture.
已经开发出一种在强阳离子交换柱 (SCX) 上的 HPLC 方法，用于分析来自不同化学基团的选定生物碱。在不同的洗脱液系统中，检查了保留、分离选择性、峰的对称性和系统效率，这些洗脱液系统含有不同类型或浓度的缓冲液，在不同的 pH 值下，并添加有机改性剂：甲醇 (MeOH)、乙腈 (CH3CN)、四氢呋喃 (THF) 或二恶烷 (Dx)。研究了保留因子作为缓冲液浓度、流动相 pH 和洗脱液中改性剂百分比的函数。在含有乙腈或四氢呋喃的洗脱液中获得了更对称的峰和最高的理论塔板数。在大多数情况下，缓冲液浓度的增加导致生物碱保留的减少、峰的对称性的改善和理论塔板数的增加。在强酸性 pH 值下的缓冲液系统中，观察到大多数被研究的生物碱的峰对称性和系统效率的提高。所得结果还揭示了用于缓冲液制备的盐阳离子的大影响。在 SCX 柱上获得的结果与在 C18 柱上获得的结果进行了比较。最有效和选择性的系统用于分离生物碱标准混合物。
