Effect of mobile phase additives on peptide retention in reversed-phase chromatography with pellicular and totally porous sorbents.
Effect of mobile phase additives on peptide retention in reversed-phase chromatography with pellicular and totally porous sorbents.
复制标题
流动相添加剂对薄膜和全多孔吸附剂反相色谱中肽保留的影响。
DOI：
10.1016/0021-9673(95)01019-x
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Horváth,C
中科院分区：
文献类型：
--
作者：
Corradini,D;Kalghatgi,K;Horváth,C
用Hy-Tach C18微膜和Vaporac C4和C18全孔固定相研究了三氟乙酸和磷酸两种移动的添加剂对肽类化合物反相色谱保留能的影响。还研究了填充有微膜吸附剂的柱的相对低的相比率的影响。在实际的洗脱液强度范围内，用不同的色谱柱和添加剂评价了两种模型肽Ac-RGGGLGLGLK-酰胺和Ac-RGAGGLGLK-酰胺的对数保留因子。在所有情况下，对数保留因子对移动的相中乙腈浓度的依赖性均呈线性。在Hy-Tach C18柱的情况下，保留对有机改性剂浓度的较高灵敏度归因于该柱的相对低的相比。对数保留因子的成对图呈线性。用两种添加剂获得的数据的曲线具有单位斜率，因此揭示了均能保留行为。另一方面，在两个不同的柱上获得的数据显示了同能保留，图的斜率不一致。分析得到了一致的结果，并验证了保留自由能可以分为两个分量所产生的移动的相和固定相的贡献的假设。该方法还允许估计柱的相对相位比，并且发现Vtaganac C18柱具有分别比Vtaganac C4和Hy-Tach C18柱高3倍和8倍的相位比。
The effect of two mobile phase additives, trifluoroacetic acid and phosphoric acid, on the energetics of peptide retention in reversed-phase chromatography was investigated using Hy-Tach C18micropellicular and Vydac C4and C18totally porous stationary phases. The effect of the relatively low phase ratio of columns packed with micropellicular sorbents was also examined. The logarithmic retention factors, of two model peptides, Ac-RGGGGLGLGK-amide and Ac-RGAGGLGLGK-amide, were evaluated with different columns and additives in a practical range of eluent strength. The dependence of the logarithmic retention factor on the concentration of acetonitrile in the mobile phase was linear in all cases. The higher sensitivity of the retention to the organic modifier concentration in the case of the Hy-Tach C18column is attributed to the relatively low phase ratio of this column. Pairwise plots of the logarithmic retention factors were linear. The plots of data obtained with the two additives has unit slopes and thus reveal homoenergetic retention behavior. On the other hand data obtained on two different columns manifest homooenergetic retention, the slopes of plots are different from unity. The analysis has yielded consistent results and validated the assumption that the retention free energy can be divided into two components arising from mobile phase and stationary phase contributions. The approach also allowed an estimation of the relative phase ratios of the columns and the Vydac C18column was found to have an 3 and 8 times higher phase ratio than the Vydac C4and the Hy-Tach C18column, respectively.