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AbstractCarnitine has key functions in energy metabolism. In most studies in animals or humans, carnitine supplementation did not influence muscle composition and function, but the effects of the carnitine esters acetylcarnitine or propionylcarnitine are not well characterized. We therefore investigated the influence of carnitine, acetylcarnitine and propionylcarnitine on body carnitine homeostasis, energy metabolism and physical performance in mice. Animals were orally supplemented with approximately 2 mmol/kg/d carnitine, acetylcarnitine or propionylcarnitine for four weeks, before being subjected to exhaustive exercise and analysis of skeletal muscle energy metabolism and morphology.In supplemented groups, the total plasma and urine carnitine concentrations were significantly higher than in the control group, whereas the skeletal muscle carnitine content remained unchanged. The bioavailability of supplemented carnitine or acylcarnitines was in the range of 20 to 23% as determined by urinary excretion of total carnitine. The supplemented acylcarnitines were hydrolyzed in the intestine and in the liver before reaching the systemic circulation. Skeletal muscle morphology including fiber type composition was not affected by treatment with carnitine or acylcarnitines. Oxygen consumption by muscle fibers obtained from soleus or gastrocnemius was not different between the groups. The supplementation of carnitine or acylcarnitines had no significant impact on the running capacity and on key substrates of skeletal muscle energy metabolism such as glycogen, ATP, phosphocreatine or creatine.In conclusion, oral supplementation of carnitine, acetylcarnitine or propionylcarnitine is associated with increased plasma concentrations of total carnitine and increased urinary excretion of carnitine, but does not affect the skeletal muscle carnitine content. Accordingly, physical performance, skeletal muscle energy metabolism, and muscle fiber type composition are not affected by high oral doses of carnitine or acylcarnitines in mice.