L-Carnitine (L-C) transports fatty acids into mitochondria for oxidation and is marketed as a weight loss supplement. In a double-blind investigation to test the weight loss efficacy of L-C, 36 moderately overweight premenopausal women were pair matched on Body Mass Index (BMI) and randomly assigned to two groups (N = 18). For 8 weeks the L-C group ingested 2 g twice daily of L-C, while the placebo (P) group ingested the same amount of lactose. All subjects walked for 30 min (60-70% maximum heart rate) 4 days/week. Body composition, resting energy expenditure (REE) and substrate utilization were estimated before and after treatment. For the subjects who completed the study (15 P, 13 L-C), no significant changes in mean total body mass (TBM), fat mass FM, and resting lipid utilization occurred over time, nor were there any significant differences between groups for any variable. Conversely REE increased significantly for all subjects, but no between group differences existed. Five of the L-C group experienced nausea or diarrhea and consequently did not complete the study. Eight weeks of L-C ingestion and walking did not significantly alter the TBM or FM of overweight women, thereby casting doubt on the efficacy of L-C supplementation for weight loss.