The purpose of this study was to investigate the effects of gender on the thermic effect of food and substrate oxidation rate during 5 hours after a mixed meal. Twenty healthy college students (10 males and 10 females) aged 20-26 years participated in this study. The energy contents of the experimental diets were 775 kcal and 627 kcal for males and females respectively, which were 30% of individual energy requirements and were composed of 65/15/20% as the proportion of carbohydrate/protein/fat. Resting and postprandial energy expenditure and substrate oxidation rates were measured with indirect calorimetry in the fasting state and every 30 min for 5 hours after meal consumption. Thermic effects of food expressed as ${Delta}AUC$ and TEF% were not significantly different between males and females. However, TEF% adjusted for body weight and fat-free mass in males (0.095% and 0.120%) were significantly lower than those in females (0.152% and 0.213%)(p < 0.05). The total amount of carbohydrate oxidized was significantly lower in males than that in females (58.6 vs. 86.6 mg/kcal energy intake/5 h, p < 0.05). In contrast, the total amount of fat oxidized was significantly higher in males than that in females after the meal (32.9 vs. 17.2 mg/kcal energy intake/5 h, p < 0.01). These results indicate that gender affects the thermic effects of food and the substrate oxidation rate after a meal. The results show that males use relatively less carbohydrate and more fat as an energy source after a meal than that of females.