Urea kinetic modeling depends critically on the parameters of the model used. When urea is removed during hemodialysis, the kinetic model is quite complex. This experiment describes for the first time the use of injected stable isotope-labeled urea to define kinetics in ESRD patients and compares the magnitude of the two urea compartments in these patients with those of control subjects. Such an experimental approach provides the kinetic data in the most direct manner. A gas chromatograph/mass spectrometer-based assay provided quantitation to as little as 0.2 mol% excess urea. The rapidly equilibrating fraction of the two urea compartments is quantified as 41.2% in ESRD patients and 33.4% in controls (P = 0.24). The rest is in a more slowly equilibrating pool. The urea clearances between these two compartments were near 1 L/min for both sets of subjects. The elimination of urea was due to the metabolic removal of the 15N label in both groups of subjects as well as renal elimination in the controls. The nearly threefold larger clearance (Cl) of labeled urea removal in controls (Cl = 74.6 mL/min) than in ESRD patients (Cl = 25.4 mL/min; P = 0.015) shows the extent to which renal clearance is more important than metabolism. These direct analyses of the fractional volumes and intercompartmental clearances for urea agree closely with previous measurements during high-efficiency hemodialysis and indicate that ESRD patients do not differ significantly from control subjects in these parameters.