It has been suggested that beta-carotene itself is unstable under certain conditions and that a combination of antioxidants may prevent the pro-oxidative effects of beta-carotene. Thus, the present study aimed to investigate the interaction of beta-carotene with three flavonoids-naringin, rutin and quercetin-on DNA damage induced by ultraviolet A (UVA) in C3H10T1/2 cells, a mouse embryo fibroblast. The cells were preincubated with beta-carotene and/or flavonoid for 1 h followed by UVA irradiation, and DNA damage was measured using comet assay. We showed that beta-carotene at 20 microM enhanced DNA damage (by 35%; P<.05) induced by UVA (7.6 kJ/m2), whereas naringin, rutin and quercetin significantly decreased UVA-induced DNA damage. When each flavonoid was combined with beta-carotene during preincubation, UVA-induced cellular DNA damage was significantly suppressed and the effects were in the order of naringin > or = rutin > quercetin. The flavonoids decreased UVA-induced oxidation of preincorporated beta-carotene in the same order. Using electron spin resonance spectroscopy, we showed that the ability of these flavonoids to quench singlet oxygen was consistent with protection against DNA damage and beta-carotene oxidation. All three flavonoids had some absorption at the UVA range (320-380 nm), but the effects were opposite to those on DNA damage and beta-carotene oxidation. Taken together, this cell culture study demonstrates an interaction between flavonoids and beta-carotene in UVA-induced DNA damage, and the results suggest that a combination of beta-carotene with naringin, rutin or quercetin may increase the safety of beta-carotene.