The effect of reactive oxygen species (ROS) on the formation of $N^{varepsilon}$-(carboxymethly)lysine (CML). one of the endproducts in the Maillard reaction of protein (or glycation), was investigated. Glyoxal, a main precursor of CML formation, was produced from both glucose and fructose during their autoxidation. The transition metal ion showed to involve in the formation of glyoxal by the metal catalyzed oxidation, suggesting that ROS accelerated the reducing sugar autoxidation. The stimulative effect of ROS on the autoxidation was more prominent in glucose than in fructose. Polyunsaturated acids (PUFAs) were shown to form glyoxal by peroxidation in proportion to the degree of unsaturation, but ROS did not affect on PUFA peroxidation. Ascorbic acid also lysine (CMHL) in the model system using hippuryl lysine and glucose had a significant effect on ROS, whereas it had no effect on ROS using glyoxal as a reactant. Almost the same trend was obtained by the analysis of antigen coated indirect noncompetitive ELISA using monoclonal antibody (6D12). These data indicated that ROS affected glucose autoxidation as well as mediated both CML and glyoxal formation, but did not affect the reactive compounds such as fructose, PUFAs and ascorbic acid.