Disinfection byproduct (DBP) formation is a potential concern with regard to MPUV/Cl2 application in water treatment. In this study, five typical amino acids (AAs) were selected to investigate their DBP alteration during short-term medium pressure (MP) UV/chlorine oxidation following post-chlorination relative to parallel dark controls. The five selected AAs include two potent DBP precursors (aspartic acid and tryptophan), one modest precursor (asparagine) and two poor precursors (phenylalanine and proline). MPUV/chlorine increased the total DBP formation and DBP-associated cytotoxicity of the two poor precursors phenylalanine (Phe) and proline (Pro) as well as their chlorine demands. Conversely, DBP formation and DBP-associated cytotoxicity of the three modest-to-potent DBP precursors showed the opposite changing trends due to MPUV/Cl2 oxidation. The two aromatic AAs (tryptophan and phenylalanine) were more readily to be affected by MPUV/Cl2 oxidation especially at acidic pH condition. Conversely, DBP formation and DBP-associated cytotoxicity of the three modest-to-potent precursors showed the opposite changing trends due to MPUV/Cl2 oxidation. Among the measured DBPs, the absolute formation potential changes of haloacetic acids and haloacetonitriles were the most prominent. Presence of bromide increased the trihalomethane formation potential of five AAs. Ammonia-spiked samples resulted in notably higher chlorine demands but slightly reduced DBPFP. Photonitration caused increased haloacetonitrile and trichloronitromethane formation but lower overall DBP formation potential and DBP-associated cytotoxicity. Results indicated that increased DBP formation of unreactive aromatic AAs may be problematic with respect to MPUV/Cl2 application, while the presence of inorganic ions may not contribute to further increase in calculated cytotoxicity of measured DBPs.