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Following the idea of green chemistry, especially green analytical chemistry, a series of stationary phases was synthesized. The obtained materials connect polar and hydrophobic groups in the structure of bonded ligands. These specific surface properties provide the stability of the stationary phase in pure water as a mobile phase. To confirm the solvation ability in purely aqueous mobile phases, excess isotherms of water and acetonitrile were determined. Further, the mixtures of nucleosides, nucleic bases and purine alkaloids were applied to test the separation selectivity of stationary phases in purely aqueous conditions at ambient temperature without any additives to the mobile phase. Among the four tested stationary phases, it is possible to find one for separation of each group of analytes that offers selective separation in reasonable time. The presented data confirms that it is possible to synthesize stationary phases for the separation of target mixtures in pure water conditions. [ABSTRACT FROM AUTHOR]