N-acetyl cysteine (NAC) is a widely used therapeutic and involved to stimulate glutathione synthesis. Glutathione elevates detoxification and works directly as a free radical scavenger. Here, we synthesized and demonstrated [1-13C]NAC as a promising novel probe for hyperpolarized 13C MRI methodologies, which has limitations in available number of probes. In vivo hyperpolarized NAC was broadly distributed throughout the body. The chemical conversions into products were observed in pancreatic tumor xenografts, Hs766t, and SU.86.86, with various conversion efficiencies depending on metabolic characteristics and status. Hyperpolarized NAC can provide insights into redox status, metabolic profile, and enzymatic activities.