Arginase-2 is upregulated following renal injury. This study used hyperpolarized pyruvate, arginine, citrulline and γ-glutamylglycine to probe changes in renal metabolism following ischemia-reperfusion in mice deficient in renal arginase-2. The kidney is a major site of arginine synthesis, and conversion of hyperpolarized citrulline to arginine can be detected, as can a signal consistent with argininosuccinate. While metabolic differences in mice lacking arginase-2 compared to controls are not readily apparent by this method, ischemia has a clear effect on several metabolite signals. Of the pyruvate metabolites, conversion to aspartate is significantly diminished, as is arginine and argininosuccinate production from citrulline.