5,5-Dimethyl-1-pyrroline-N-oxide (DMPO) is used to detect reactive oxygen species in vitro, and N-acetyl-L-cysteine (NAC) is an antioxidant. We synthesized 13C-labeled DMPO and NAC, and investigated feasibility of hyperpolarized 13C-DMPO and 13C-NAC for evaluating oxidative stress in mice. Hyperpolarized 13C-DMPO and 13C-NAC provided a single peak at 76 ppm and 174 ppm, respectively, and the T1 relaxation time was sufficiently long to apply them for mouse imaging. The signals 13C-DMPO and 13C-NAC were also detected in a mouse body after intravenous injection. The results showed 13C-DMPO and 13C-NAC can be applied to some disease models to evaluate oxidative stress in vivo.