The effects of ferrocenyl chalcones including 1-acetyl-1′-(m-methoxy-p-hydroxycinnamoyl)ferrocene (AVF), 1,1′-di(m-methoxy-p-hydroxycinnamoyl)ferrocene (DVF), and 1-acetyl-1′-(m, p-dihydroxycinnamoyl)ferrocene (APF) on the oxidative damage of DNA were explored when glutathione (GSH) and Cu2+, hydroxyl radical (•OH), and 2,2′-azobis(2-amidinopropanehydrochloride) (AAPH) acted as the oxidation-initiators. These ferrocenyl chalcones functioned as prooxidants in Cu2+/GSH- and •OH-induced oxidations of DNA. Ferrocene itself promoted AAPH-induced oxidative damage of DNA, and 1,1′-dicinnamoylferrocene (DCF) showed very weak inhibitive effect on AAPH-induced oxidation of DNA. On the other hand, APF, DVF, and AVF were able to trap 5.3, 5.5, and 5.7 radicals, respectively, in protecting DNA against AAPH-induced oxidation. Therefore, ferrocenyl chalcones were effective antioxidants to protect DNA against peroxyl radical-induced oxidation.