Aggregation studies were performed on platelets from five patients with Bartter's syndrome. Epinephrine failed to induce aggregation in all five patients. Adenosine 5'-diphosphate (ADP) produced a single reversible phase of aggregation, and there was depressed sensitivity to collagen. Response to ristocetin was normal. There was a dose-related inhibition of ADP-induced platelet aggregation when plasma from the patients was added to normal platelet-rich plasma. This inhibition was diminished or absent when patients were receiving aspirin. Washed platelets from two patients who were no longer undergoing aspirin therapy, showed a normal response to epinephrine in normal platelet-poor plasma. Bleeding time was reduced from 23 minutes to 12 minutes in one patient while on aspirin therapy. These studies suggest that a circulating inhibitor of platelet aggregation, probably of prostaglandin origin, is present in the plasma of patients with Bartter's syndrome.