The purpose of this study was to determine whether submaximal exercise significantly changes the concentration of vitamin E (αToc) in rat liver and skeletal muscle and to establish a time course for the return to basal levels. Male Sprague-Dawley rats, age 8 to 10 weeks, were randomly divided into sedentary control (Con) (n = 7) and exercise n = 17) groups. Exercised animals ran 100 min on a motorized treadmill at approximately 70% VO2max for 3 consecutive days. They were then sacrificed immediately postexercise (0Post), 24 hr post (24Post), or 72 hr post (72Post). The gastrocnemius, red vastus lateralis (RV), white vastus lateralis (WV), and liver were excised and analyzed for αToc concentration by high-performance liquid chromolography utilizing electrochemical detection. We found that after 3 consecutive days of exercise, αToc was reduced in RV and WV at 0Post and 24Post but returned to control values by 72Post. Liver αToc content was not changed at OPost but was significantly reduced at 24 Post and 72 Post. No significant changes in αToc were observed in the gastrocnemius in response to exercise. The data indicate that following an exercise-related decrease, skeletal muscle vitamin E concentration requires more than 24 hr to return to the preexercise concentration, and that the replenishment process may involve redistribution of vitamin E from liver to muscle.