The thin-layer chromatographic behavior of cholic acid and fourteen of its synthetic derivatives has been studied in six reversed-phase and three normal-phase systems. Four significantly different adsorbents were used — RP-18-silica, CN-silica, polyacrylonitrile adsorbent, and unmodified silica gel. Reversed-phase chromatography was performed with water-organic modifier (methanol, dioxane, or acetone) binary mobile phases of widely variable composition. An approximately linear relationship was obtained between RM values and the amount of organic modifier in the mobile phases investigated. The effect of mobile-phase composition on retention has been considered and the selectivity of the chromatographic systems is also discussed. Separation mechanisms are proposed on the basis of the results obtained. A positive solvation effect was proposed as a predominant factor determining retention under normal-phase conditions whereas hydrophobic interactions of the substances with non-polar parts of the adsorbents were assumed to be predominant under reversed-phase conditions.
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