Separation of eburnane alkaloids using reversed-phase high-performance liquid chromatography was investigated on muBondapak C18 in LiChrosorb RP-8 columns with acetonitrile-aqueous 0.01 M ammonium carbonate as eluent. The method can be successfully applied for the group separation of eburnane alkaloids as well as for the separation of stereoisomers and of ester homologues.