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Three novel norfloxacin-based ruthenium(ii) complexes, [Ru(bpy)2(NFLX)]Cl·2H2O (1), [Ru(phen)2(NFLX)]Cl·2H2O (2), and [Ru(dmbpy)2(NFLX)]Cl·2H2O (3) (bpy = 2,2′-bipyridine, phen = 1,10-phenanthroline, dmbpy = 4,4′-dimethyl-2,2′-bipyridine, and NFLX = norfloxacin), were synthesised and characterised with electrospray ionisation mass spectrometry and 1H and 13C NMR spectroscopy. The antitumour properties were evaluated by MTT assay, and the data revealed that 2 can inhibit the growth of human lung adenocarcinoma A549 efficiently. Furthermore, the DNA-binding behaviours of these complexes were investigated by a multiple spectroscopy assay and viscosity study. The results indicated that these complexes interact with calf thymus DNA through electrostatic interactions with a strong binding affinity in the order 2 > 3 > 1. Therefore, these results suggested that 2 might be a suitable anticancer agent due to its excellent DNA-binding abilities. Three novel norfloxacin-based ruthenium(ii) complexes [Ru(bpy)2(NFLX)]Cl·2H2O (1), [Ru(phen)2(NFLX)]Cl·2H2O (2), and [Ru(dmbpy)2(NFLX)]Cl·2H2O (3) (bpy = 2,2′-bipyridine, phen = 1,10-phenanthroline, dmbpy = 4,4′-dimethyl-2,2′-bipyridine, and NFLX = norfloxacin) have been synthesised. Compound 2 exhibited a stronger DNA-binding affinity than 1 and 3 , which is consistent with its antitumour activity, indicating that the DNA-binding behaviour can be closely related to the anti-tumour activity of the ruthenium(ii) complexes. [ABSTRACT FROM AUTHOR]