Antioxidant supplements are expected to decrease oxidative damage and prevent ocular diseases. In this study, changes in the anti-oxidative ability and oxidative status in the aqueous humor before and after intake of a lutein-containing supplement were measured. Forty patients who all had identical grades of cataracts in both eyes were included. The aqueous humor was collected as pre-intake samples during cataract surgery. Ocuvite + Lutein(Ⓡ), an antioxidant supplement, was administered orally beginning the day after surgery. Six weeks later, the aqueous humor was collected as a post-intake sample during cataract surgery of the opposite eye. To determine the anti-oxidative ability, the levels of superoxide (O2(*-)) scavenging activity were measured. To determine the oxidative status, the levels of hydrogen peroxide (H2O2) and total amount of hydroperoxides (TH, including H2O2 and peroxides of lipids, proteins, and nucleic acids) were measured. In post-intake samples, the O2(*-) scavenging activities were significantly higher in both genders (p < 0.05). The levels of H2O2 were significantly higher (p < 0.01) while the levels of TH were significantly lower (p < 0.01) only in females. The level of H2O2 was significantly negatively correlated to the TH in the post-intake samples of both genders (r = -0.50 and p < 0.05 for males; r = -0.59 and p < 0.01 for females) while the level of H2O2 was significantly positively correlated to the O2(*-) scavenging activity in both pre- and post-intake aqueous humor in females only (r = 0.66 and p < 0.01 for pre-intake samples, r = 0.71 and p < 0.01 for post-intake samples). After the introduction of the antioxidant supplement, O2(*-) scavenging activity increased while H2O2 levels remained the same in males, suggesting that scavenging rates are proportional. In postmenopausal females, the superoxide scavenging activity also increased, however unlike in males, the H2O2 levels also increased meaning H2O2 was not completely scavenged. H2O2 is a non-free radical and can be excreted from the aqueous humor to prevent further oxidation of lipids, proteins, and nucleic acids, which was confirmed by the low levels of TH in post-intake samples. Antioxidant supplements are suggested to be effective in reducing oxidation in the aqueous humor by different mechanisms in both genders.