Enzymatically prepared split frog skins consisted purely of epithelial cells. Electrical parameters and the cell contents of ATP, ADP, phosphocreatine (PCr), creatine, inorganic phosphate, protein, and water were measured in skins maintained at room temperature. Studies were conducted under base-line conditions, 15 and 60 min after adding vasopressin, and 30 min after adding amiloride. Intracellular ionic activities and concentrations were obtained from previous results. The data demonstrated that 1) the base-line concentration ratio of PCr/ATP was 0.53 +/- 0.03; 2) the average molar free energy of hydrolysis of intracellular ATP was approximately 15.0 kcal X mol-1 under control conditions, changing by less than or equal to 3% with changes in transport; and 3) the free energy of extruding 3 mol of Na+ and accumulating 2 mol of K+ was approximately 9.8 kcal X mol-1 under base-line conditions; the difference between the molar free energies of ATP hydrolysis and of transport work remained large, despite large changes in transepithelial transport. The simplest conclusion is that the Na+ pump of frog skin operates far from equilibrium.