2H-MRS is a novel method of imaging flux through metabolic pathways. Here, we show that 2H-MRS can be used to probe the isocitrate dehydrogenase 1 mutation (IDHmut), which catalyzes production of the oncometabolite 2-hydroxyglutarate (2-HG) in low-grade gliomas. Our results indicate that 2-HG production from [3,3’-2H]-α-KG can be specifically observed in IDHmut cells. Importantly, treatment with an IDHmut inhibitor, which is in clinical trials for low-grade glioma patients, results in a significant reduction in 2-HG production from [3,3’-2H]-α-KG in IDHmut cells. Our results point to the potential of [3,3’-2H]-α-KG as a probe of glioma IDHmut status and response to therapy.