We investigated the possible role of enterohepatic recirculation in prolongation of the half-life of elimination for Adriamycin, a commonly prescribed anticancer agent. We sought to determine whether enterohepatic recirculation of Adriamycin and its metabolites occurs using a linked-rat model. Two rats, a donor and a receiver, were linked via a catheter from the bile duct of the donor rat to the duodenum of the receiver. Control experiments were conducted with intact rats (without a bile duct cannula, control A) in order to estimate the half-life of elimination and with bile duct-cannulated rats (control B) to determine the amounts of Adriamycin and its metabolites in the bile. [14C-14]-Adriamycin was injected intravenously via the femoral vein to control A, control B and donor rats. The biological half-life of Adriamycin in the intact rats (control A, 10 h) was significantly higher than in the bile-duct-cannulated rats (control B, 4 h). The cumulative amount of Adriamycin and its metabolites excreted in the urine of the control A rats was also greater than from control B rats, indicating higher levels of the drug in their systemic circulation. Biological samples (bile, urine, plasma, blood cells and the major organs heart, liver and kidney) of the receivers contained significant amounts of Adriamycin and its metabolites. The total radioactivity recovered in the bile of the receivers accounted for 0.1% to 8% of the Adriamycin dose that was administered to the donors. Adriamycin and its metabolites appeared there only after a lag time that was consistent among all the receivers. Doxorubicinol aglycone was the major metabolite found in the bile and urine of the receivers. Low but constant levels of radioactivity were also detected in the plasma and blood cells of the receivers. The presence of unchanged Adriamycin in the bile and urine of the receivers suggested absorption of the parent drug from the intestine of the receivers. Overall, we estimated that about 22% of the dose injected to the donors was absorbed from the intestine of the receivers. Taken together, these findings clearly demonstrate a significant role for enterohepatic recirculation of Adriamycin and its metabolites, which may contribute to the ability of these compounds to induce cumulative cardiac damage and/or to increase the efficacy of Adriamycin.