In the present study we investigated the relationship between plasma fatty acids (FA) and intramuscular triacylglycerol (IMTAG) kinetics of healthy volunteers. To this aim [U-13C]-palmitate was infused for 10 hours and FA kinetics determined across the leg. In addition, the rate of FA incorporation into IMTAG in vastus lateralis muscle was determined during two consecutive 4-h periods (2-6 h and 6-10 h). Fifty to 60% of the FA taken up from the circulation were esterified into IMTAG, whereas 32-42% were oxidized during between 2-6 and 6-10 h. IMTAG fractional synthesis rate was 3.4±0.8%h-1 and did not change between the two 4-hours periods, despite an increase in arterial FA concentration (34%, p<0.01). IMTAG concentration was also unchanged, implying that the IMTAG fractional synthesis rate was balanced by an equal rate of breakdown. FA oxidation increased over time, which could be due to the observed decline in plasma insulin concentration (-74 %, p<0,01). In conclusion, a substantial fraction of the fatty acids entering skeletal muscle in post-absorptive healthy individuals is esterified into IMTAG, due to its high turnover rate (29 h.pool-1). An increase in FA level, as a consequence of short-term fasting, does not seem to increase IMTAG synthesis rate and pool size.