Little is known regarding the effects of mineral-containing supplemention on oxidative stress markers, carbohydrate and lipid metabolism in response to an acute bout of exercise in humans. The present study investigated whether prior mineral-containing supplemention can improve oxidative stress status and how this supplementation influences on carbohydrate and lipid metabolism after a single bout of cycling in young men. Twelve, healthy young men (aged 22.5 ± 2.4 years, mean ± SD) underwent two, 150-min trials in a random order. Each participant received oral administration of mineral supplement containing 13.4 mg of sulphur or placebo one minute before exercise. In both trials, participants cycled at 75% of heart rate reserve for 60 minutes and then rested for 90 minutes. Blood samples were collected pre-exercise supplementation, immediately after exercise, 30 minutes after exercise, 60 minutes after exercise and 90 minutes after exercise. Circulating concentrations of derivatives of reactive oxygen metabolites, biological antioxidant potential, glucose and insulin did not differ between trials. Elevated circulating concentrations of non-esterified fatty acids were observed immediately after exercise in the supplementation trial compared with the placebo trial (mean ± SD: 1.1 ± 0.5 and 0.9 ± 0.3 mmol/L, respectively: trial × time interaction, p = 0.036). The present study showed that acute mineral-containing antioxidant supplemention appears to have no effect on improving oxidative stress markers in response to a single bout of cycling in healthy young men. In addition, the findings of this study suggested that mineral-containing supplemention may influence lipids metabolism during exercise.