Cation-exchange stationary phases were characterized in different chromatographic modes (HILIC, RPLC, IC) and applied to the separation of non-charged hydrophobic and hydrophilic analytes. The set of columns under investigation included both commercially available cation-exchangers and self-prepared PS/DVB-based columns, the latter consisting of adjustable amounts of carboxylic and sulfonic acid functional groups. The influence of cation-exchange site and polymer substrate on the multimodal properties of cation-exchangers was identified using selectivity parameters, polymer imaging and excess adsorption isotherms. Introducing weakly acidic cation-exchange functional groups to the unmodified PS/DVB-substrate effectively reduced hydrophobic interactions, whilst a low degree of sulfonation (0.09 to 0.27% w/w sulphur) mainly influenced electrostatic interactions. Silica substrate was found to be another important factor for inducing hydrophilic interactions. The presented results demonstrate that cation-exchange resins are suitable for mixed-mode applications and offer versatile selectivity.