1. Whole-body kinetics and regional exchange of glycerol across forearm muscle were assessed in eight lean subjects by a combination of a tracer method (infusion of [2H5]glycerol) and arteriovenous catheterization. 2. During an apparent steady state, the enrichment of glycerol in deep venous blood from the muscle bed of the forearm was about half (4.40 +/- 1.72 atom per cent excess) that observed in arterialized blood (8.41 +/- 4.30 atom per cent excess). Under the same conditions, the circulating concentrations of glycerol in arterialized (91 +/- 24 mumol/l) and venous (87 +/- 32 mumol/l) blood were similar. 3. In a further group of 37 subjects it was found that about half had a positive arteriovenous concentration difference and the other had half a negative arteriovenous concentration difference (mean -1.6 +/- 11.9 mumol/l; range -25 to +22 mumol/l). 4. These results suggest: (a) that human muscle does not always release glycerol and may take it up; (b) that there is substantial isotopic exchange of glycerol across forearm muscle tissue, which is not reflected by the net exchange of glycerol; this could be due to slow equilibrium of enriched glycerol from the circulation, with unenriched free glycerol in the muscle pool, or due to the simultaneous metabolic utilization of enriched glycerol and metabolic production of unenriched glycerol; (c) that the estimation of glycerol flux rates is strongly dependent on whether the blood is arterialized or deep venous.