Bendavia is a mitochondria‐targeted peptide that protects the heart against ischemia‐reperfusion (I/R) injury in a wide range of experimental models. We have previously shown that Bendavia's mechanism of action involves lowering intracellular levels of reactive oxygen species (ROS). As other compounds that lower ROS levels can abrogate the protective effects of ischemic preconditioning (IPC), this study was conducted to determine if Bendavia influenced the protection evoked by IPC. Isolated guinea pig hearts were put into one of 4 groups: untreated controls (n=7), perfusion with 1nM Bendavia without IPC (n=5), untreated+IPC (n=6), and IPC+1nM Bendavia (n=5). All hearts were exposed to I/R (20/120 min respectively), with cardiac electromechanical function monitored throughout the protocol. Infarct size (expressed as % of zone‐at‐risk; ZAR) was reduced in hearts treated with Bendavia (29 +/− 2.4% ZAR) when compared to untreated hearts (42% +/−2.5 P=<0.01 ). IPC also reduced infarct size to 26 +/−3.3% ZAR, and the protective effect of IPC was not influenced by Bendavia (23 +/− 1.6% ZAR). We conclude that Bendavia may be an attractive candidate for preventing cardiac injury without influencing endogenous protective signaling. Funding for this project included support from the APS UGSRF program.