Puberty associated with intense physical activity results in oxidation stress. Zinc supplementation may benefit antioxidant capacity although it may also affect iron and copper status. This study evaluated the effect of zinc supplementation on antioxidant, zinc and copper status of physically active male football players (13 years +/- 0.4 years), divided in two groups and studied during 12 weeks: Zn-supplemented (Zn-SUP, 22 mg Zn d(-1) as zinc gluconate, n = 21) and placebo (PLA, n = 26). At baseline, there was no significant difference in biochemical indices between the two groups. After treatment, plasma zinc and erythrocyte iron increased in both groups (p < 0.001); urinary zinc increased (p < 0.001) only in Zn-SUP, and erythrocyte zinc decreased (p = 0.002) only in PLA. Plasma iron and copper decreased (p = 0.01 and p = 0.015, respectively) only in Zn-SUP. Plasma ferric-reducing ability and plasma conjugated dienes increased, and erythrocyte osmotic fragility decrease in both groups, although the latter two were significantly lower in Zn-SUP compared to PLA (p < 0.01). In conclusion, our study indicates that the use of 22 mg d(-1) of supplemental zinc during 12 week in adolescent athletes did not affect growth, improved markers of antioxidant status but reduced plasma iron and copper. Therefore, it appears that the use of zinc supplementation by healthy adolescent athletes benefits their antioxidant capacity but impairs copper and iron nutritional status.