The retention mechanism and separation of various aromatic sulfonic and carboxylic acids on reversed-phase liquid chromatographic column were studied in the mobile phase containing dodecyltrimethylammonium bromide (DTAB). The retention mechanism was found to be followed the ion-interaction model where the DTAB occupies a primary layer at the stationary phase while the sample anions and other co-anions in the system compete for forming the secondary layer. The capacity factors of samples were influenced by the several factors such as pH, concentration of various organic solvents, co-anions in the mobile phase and functional groups in sample molecules. Some mixtures of organic samples were attempted to separate under optimum condition.