Efficiency of transient supramolecular enantioselective recognition processes during direct, isocratic RP-HPLC separation of all four stereoisomers (SR,RS,SSandRR) of labetalol, chemically known as 2-hydroxy-5-[1-hydroxy-2-((1-methyl-3-phenylpropyl) amino)ethyl] benzamide, an antihypertensive agent with combined alpha- and beta-adrenoceptor blocking activities, have been presented. In the developed RP-HPLC system the conventional octadecylsilica packed column and double-helical chelate (-)(M)(λ,Λ)-[4,4'-(1S)-methyl-(2R)-propylethane-diyldi-imino) bis(pent-3-en-2-onato)]nickel(II) as the chiral mobile phase additive was used.