We examined the relationship between tricarboxylic acid (TCA) cycle intermediate (TCAI) pool size, TCA cycle flux (calculated from leg O2 uptake), and pyruvate dehydrogenase activity (PDHa) in human skeletal muscle. Six males performed moderate leg extensor exercise for 10 min, followed immediately by intense exercise until exhaustion (3.8 +/- 0.5 min). The sum of seven measured TCAI (SigmaTCAI) increased (P </= 0.05) from 1.39 +/- 0.11 at rest to 2. 88 +/- 0.31 after 10 min and to 5.38 +/- 0.31 mmol/kg dry wt at exhaustion. TCA cycle flux increased approximately 70-fold during submaximal exercise and was approximately 100-fold higher than rest at exhaustion. PDHa corresponded to 77 and 90% of TCA cycle flux during submaximal and maximal exercise, respectively. The present data demonstrate that a tremendous increase in TCA cycle flux can occur in skeletal muscle despite a relatively small change in TCAI pool size. It is suggested that the increase in SigmaTCAI during exercise may primarily reflect an imbalance between the rate of pyruvate production and its rate of oxidation in the TCA cycle.