The purpose of present study was to evaluate the effect of endurance exercise training on skeletal muscle free amino acid concentrations, and differences in free amino acid concentration between soleus muscle which consists of mostly slow twitch oxidative fiber and extensor digitorum longus muscle which consists of fast twitch oxidative glycolytic fiber. Sixteen male SD rats (4 weeks old) were randomly devided into two groups, and fed a purified AIN-93M diet with or without aerobic exercise training according to the protocol (running on the treadmill at 25 m/min for 60 min, 5 days a week) for 6 weeks. Exercise-training for 6 weeks significanly reduced the commulative body weight gain (p＜0.05) and food efficiency ratio (p＜0.01) of rats. The result showing mitochondrial citrate synthase activity of soleus muscle was significantly higher in exercise-trained rats compared to the value for control animals (p＜0.01) indicates aerobic exercise-training was successfully accomplished in the trained group. No difference was found in the muscle aminogram pattern between soleus muscle and extensor digitorum longus muscle of control animals. However, free amino acid concentrations of soleus muscle were from 1.2 to 3.9 times of those found in extensor digitorum longus muscle of control rats, depending on an individual amino acid. Intermediate level of endurance exercise training for 6 weeks did not influence concentrations of most of free amino acid in soleus muscle of rats collected at an overnight fasted and rested state. In contrast, isolucine and leucine concentrations in extensor digitorum longus muscle of exercise-trained rats were significantly lower than those for control animals. These results indicate that aerobic energy metabolism had not been efficiently conducted, and thereby the utilization of BCAA for energy substrate was enhanced in fast twitch oxidative glycolytic fibers of extensor digitorum longus muscle of rats followed exercise-training protocol for 6 weeks.