These experiments were designed to examine the mechanisms involved in the renal excretion of the non-nutritive sweetener, saccharin. Renal transport of saccharin in female rats was quantitatively evaluated using renal cortical slices in vitro and renal clearances in vivo. Renal cortical slices actively accumulated saccharin. Accumulation was oxygen dependent, saturable and reduced in the presence of metabolic inhibitors (2,4-dinitrophenol and sodium azide) and other organic anions 1p-aminohippurate (PAH) and probenecid]. Furthermore, addition of acetate or lactate to the medium stimulated saccharin uptake whereas reducing potassium concentration in the medium significantly decreased saccharin accumulation. Addition of saccharin to medium containing PAH and N-methylnicotinamide produced a dose-related depression of PAH accumulation. Although N-methylnicotinamide accumulation also was reduced, the depression was not dose-related. The saccharin/inulin clearance ratio of 3.76 indicates that saccharin, like PAH, undergoes tubular secretion. These findings suggest that the primary route of renal elimination of saccharin is active tubular secretion. It is also suggested that saccharin and PAH may share a common transport system.