Effects of verapamil, an calcium channel blocker, on hepatic glutathione were studied in vivo in the rat and in the perfused rat liver. An injection of verapamil at a dose of 15 mg/kg body weight but not at 5 mg/kg significantly decreased hepatic glutathione contents in both fed and fasted animals 6 h after the injection. The administration of verapamil at a dose of 10 mg/kg twice a day for a week brought a significant decrease in hepatic glutathione contents and a significant increase in plasma glutathione levels. In the perfused rat liver, sinusoidal glutathione efflux was significantly increased when verapamil was added to the perfusion medium in a concentration of 20 microM. These data indicate that verapamil increases glutathione efflux from the liver and that calcium mobilization may be concerned in glutathione efflux in vivo.