To study whether sucrose administration acts on lipid mobilization during prolonged exercise, we used subcutaneous abdominal adipose tissue microdialysis in eight well-trained subjects submitted at random to two 100-min exercises (50% maximal aerobic power) on separate days. After 50 min of exercise, the subjects ingested either a sucrose solution (0.75 g/kg body wt) or water. By using a microdialysis probe, dialysate was obtained every 10 min from the subjects at rest, during exercise, and during a 30-min recovery period. During exercise without sucrose, plasma and dialysate glycerol increased significantly. With sucrose, the response was significantly lower for dialysate glycerol (P < 0.05). Plasma free fatty acid level was lower after sucrose than after water ingestion (P < 0.05). With water ingestion, plasma catecholamines increased significantly, whereas insulin fell (P < 0.05). With sucrose ingestion, the epinephrine response was blunted, whereas the insulin level was significantly increased. In conclusion, the use of adipose tissue microdialysis directly supports a lower lipid mobilization during exercise when sucrose is supplied, which confirms that the availability of carbohydrate influences lipid mobilization.