We measured splanchnic and leg glycerol [and free fatty acid (FFA)] uptake and release in 11 healthy volunteers before and after meal ingestion to assess whether regional FFA-to-glycerol release ratios mirror systemic release ratios. Basal splanchnic triglyceride release was also assessed. Although basal splanchnic glycerol uptake (111 +/- 18 micromol/min) accounted for most of systemic glycerol rate of appearance (156 +/- 20 micromol/min), leg glycerol uptake was also noted. The basal, systemic FFA-to-glycerol release ratio was less (2.6 +/- 0.2, P < 0.05) than the splanchnic ratio of 6.1 +/- 1.3, and the leg FFA-to-glycerol release ratio under fed conditions was less than the systemic ratio (0.9 +/- 0.1 vs. 1.6 +/- 0.2, respectively, P < 0.05). Basal splanchnic triglyceride production rates were 74 +/- 20 micromol/min, which could produce equimolar amounts of glycerol in the peripheral circulation via lipoprotein lipase action. In summary, 1) regional FFA-to-glycerol release ratios do not mirror systemic ratios, 2) leg glycerol uptake occurs in humans, and 3) splanchnic triglyceride production rates are substantial relative to systemic glycerol appearance. Glycerol appearance rates may not be a quantitative index of whole body lipolysis.