Prednisolone, atrolactic acid, and ibuprofen were separated in one HPLC run on an Obelisc R HPLC column. Obelisc R columns have C12 carbon chain and basic and acidic groups on the surface of silica gel. Compounds are separated by combination of reversed-phase, cation-exchange, and anion-exchange mechanisms. Obelisc R column can be used for retention and separation of acidic and basic hydrophilic compounds, acidic and basic hydrophobic compounds, and neutral compounds. Elution is monitored by common detection techniques like UV, ELSD, CAD and LC/MS.