Abstract <p>A method is developed for the enantioselective determination of pharmaceutical environmental pollutants (beta-blockers, profens) by high-performance liquid chromatography using promising chiral stationary phases based on eremomycin derivatives. The specificity of the studied selectors is revealed: the phase based on eremomycin aglycone is effective for separating more polar pharmaceuticals, including β-blockers, while the phase containing chloreremomycin demonstrated high selectivity for less polar profens. A possibility of the simultaneous separation of the enantiomers of substances in multicomponent mixtures is demonstrated. A method for water sample preparation using solid-phase extraction is developed and tested. An analysis of real natural and wastewater samples confirmed the applicability of the developed approach to identifying chiral pharmaceutical contaminants.</p>