It is commonly acknowledged that the retention of non-ionized polar analytes on polar stationary phases is governed by hydrophilic partitioning and surface adsorption. However, it has been difficult to evaluate whether partitioning or adsorption is the dominant mechanism for a specific polar compound on a polar stationary phase. We have developed a simple method based on the thermodynamic principle of partitioning to quantitatively investigate the retention contributed by the partitioning or adsorption mechanism. By varying phase ratio through changing salt concentration in the mobile phase, we were able to determine the distribution coefficients of cytosine between the adsorbed water layer and the mobile phase containing various levels of acetonitrile. The retention factors of cytosine attributed to partitioning and adsorption were quantitatively determined. The results demonstrate that the dominant retention mechanism for cytosine is hydrophilic partitioning on ZIC-HILIC, XBridge Amide and LUNA-HILIC columns.
Keywords: Distribution coefficients; Hydrophilic partitioning; Phase ratio; Surface adsorption.
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