The changes in the muscle contents of CoASH and carnitine and their acetylated forms, lactate and the active form of pyruvate dehydrogenase complex were studied during incremental dynamic exercise. Eight subjects exercised for 3–4 minutes on a bicycle ergometer at work loads corresponding to 30, 60 and 90% of their V_02max Muscle samples were obtained by percutaneous needle biopsy technique at rest, at the end of each work period and after 10 minutes of recovery. During the incremental exercise test there was a continuous increase in muscle lactate, from a basal value of 4.5 mmol kg^‐1 dry weight to 83 mmol kg^‐1 at the end of the final period. The active form of pyruvate dehydrogenase complex increased from 0.37 mmol acetyl‐CoA formed per minute per kilogram wet weight at rest to 0.80 at 30% V_02max1.28 and 1.25 at 60 and 90% V_02max respectively. Both acetyl‐CoA and acetylcarnitine increased at the two highest work loads. The increase of acetyl‐CoA was from 12.5 μmol kg^‐1 dry weight at rest to 27.3 after the highest work load and for acetylcarnitine from 6.0 mmol kg^‐1 dry weight to 15.2. The CoASH and free carnitine contents fell correspondingly. There was a close relationship between acetyl‐CoA and acetylcarnitine accumulation in muscle during exercise, with a binding of ˜ 500 mol acetyl groups to carnitine for each mole of acetyl‐CoA accumulated. The results imply that the carnitine store in muscle functions as a buffer for excess formation of acetyl groups from pyruvate catalyzed by the pyruvate dehydrogenase complex.