Left ventricular flow progressively decreases during preservation of ex vivo hearts by continuous hypothermic perfusion, according to previous studies. This study was done to determine whether left ventricular flow could be maintained during 24-hour hypothermic perfusion by the addition of adenosine to the perfusate. Isolated canine hearts were perfused with either a modified Krebs-Henseleit solution or with the control solution containing adenosine (20 mumol/L) for either 4 or 24 hours. On completion of the perfusion period, radioactive microspheres were injected into the arterial inflow of the hearts. Myocardial samples were then taken from the endocardial, midcardial, and epicardial portions of the free wall and papillary muscles of the right and left ventricles, as well as the interventricular septum. Concentrations of radioactivity were determined for each sample, summed, and averaged for each ventricle. The ratio of average concentrations (L/R ratio) was significantly increased in the 24-hour adenosine hearts compared with 24-hour controls (1.10 +/- 0.15 vs 0.64 +/- 0.21, p less than 0.005). The L/R ratio of 4-hour controls (1.30 +/- 0.48) was also greater than that of 24-hour controls (p less than 0.005). These results suggest that left ventricular flow decreases during continuous hypothermic perfusion. The tendency toward diminished left ventricular perfusion after 24 hours is reduced when adenosine is added to the perfusate. Adenosine may be beneficial in ex vivo heart preservation with hypothermic perfusion by improving left ventricular flow.