To ascertain the effects of hair dyeing application on oxidative stress in human, a mixture of permanent black colored hair dye with the same amount of oxidant containing 6% hydrogen peroxide was used. A hair dyeing with contacting the scalp (conventional dyeing) and a hair dyeing with 3 to 4mm away from the scalp (alternative dyeing) were applied to each 15 young healthy women. Blood was taken from the brachial vein at two sampling times, just before and 6 hours alter the hair dyeing, and antioxidant enzyme activities and antioxidant contents were measured in red blood cells. After dyeing, malondialdehyde(MDA) contents for conventional dyeing group was shown to a tendence of more increased than alternative dyeing group. After dyeing, reduced glutathione (GSH) contents for conventional dyeing group was shown to a tendence of more decreased than alternative dyeing group. After dyeing, superoxide dismutase (SOD) and catalase (CAT) activities were significantly decreased in conventional dyeing group (p < 0.01), however, SOD and CAT activities were not significantly decreased in alternative dyeing group. After dyeing, there was no significant decrease in glutathione peroxidase (GPX) activity both for conventional dyeing group and alternative dyeing group. Therefore, after dyeing, the degree of oxidative stress in red blood cells for alternative dyeing group was appeared to be lower than conventional dyeing group.