The separation of epimers of anisodamine were studied with HPLC using a reversed phase ODS C18 column. Several chiral additives were tried in the mobile phases. d-Camphorsulfonic acid and L-(+)-dibutyl tartrate showed good resolution ability for the separation of the isomers, a pair of epimers of anisodamine were well separated; while beta-cyclodextrin appeared to have no selectivity for the epimers.