This study determined if the suppression of lipolysis after preexercise carbohydrate ingestion reduces fat oxidation during exercise. Six healthy, active men cycled 60 min at 44 +/- 2% peak oxygen consumption, exactly 1 h after ingesting 0.8 g/kg of glucose (Glc) or fructose (Fru) or after an overnight fast (Fast). The mean plasma insulin concentration during the 50 min before exercise was different among Fast, Fru, and Glc (8 +/- 1, 17 +/- 1, and 38 +/- 5 microU/ml, respectively; P < 0.05). After 25 min of exercise, whole body lipolysis was 6.9 +/- 0.2, 4.3 +/- 0.3, and 3.2 +/- 0.5 micromol x kg(-1) x min(-1) and fat oxidation was 6.1 +/- 0.2, 4.2 +/- 0.5, and 3.1 +/- 0.3 micromol x kg(-1) x min(-1) during Fast, Fru, and Glc, respectively (all P < 0.05). During Fast, fat oxidation was less than lipolysis (P < 0.05), whereas fat oxidation approximately equaled lipolysis during Fru and Glc. In an additional trial, the same subjects ingested glucose (0.8 g/kg) 1 h before exercise and lipolysis was simultaneously increased by infusing Intralipid and heparin throughout the resting and exercise periods (Glc+Lipid). This elevation of lipolysis during Glc+Lipid increased fat oxidation 30% above Glc (4.0 +/- 0.4 vs. 3.1 +/- 0.3 micromol x kg(-1) x min(-1); P < 0.05), confirming that lipolysis limited fat oxidation. In summary, small elevations in plasma insulin before exercise suppressed lipolysis during exercise to the point at which it equaled and appeared to limit fat oxidation.