Although hypoxia and metabolic acidosis have both been shown to impair cardiac function, some workers have suggested that acidosis during a period of hypoxia will actually accelerate physiologic recovery from this insult. To address the interactions of metabolic acidosis and hypoxia further, isolated isovolumic rat hearts were exposed to normal perfusion conditions for 30 min to establish baseline conditions, then either continued normal conditions, metabolic acidosis, hypoxia, or combined acidosis and hypoxia for 30 min and subsequently reperfused under normal perfusion conditions for an additional 30 min. We observed that acidosis + hypoxia impaired recovery of cardiac contraction more than acidosis or hypoxia alone following experimental perfusion. The combination of acidosis and and hypoxia also impaired cardiac energy metabolism more than acidosis or hypoxia alone as assessed by increases in tissue inorganic phosphate during experimental perfusion as well as during reperfusion. These data suggest that during hypoxia, acidosis appears to primarily impair cardiac energy production as we have previously observed in the normoxic isolated rat heart. Therefore, in the intact beating heart, acidosis may not protect from hypoxic injury as has been suggested in simpler systems but may not protect from hypoxic injury as has been suggested in simpler systems but rather may exacerbate at.