Many glioma patients harbor specific mutations in the isocitrate dehydrogenase gene IDH1, which associate with a relatively better prognosis. IDH1-mutated tumors produce the oncometabolite 2-hydroxyglutarate. Since IDH1 regulates several pathways leading to lipid synthesis, we hypothesized that IDH1 mutant tumors would display an altered phospholipid metabolite profile which would impinge on tumor pathobiology. To investigate this hypothesis, we performed 31P-MRS imaging in mouse xenograft models of four human gliomas, one of which harbored the IDH1-R132H mutation. 31P-MR spectra from the IDH1-mutant tumor displayed a pattern distinct from that of the three IDH1 wild-type tumors, characterized by decreased levels of phosphoethanolamine and increased levels of glycerophosphocholine. This spectral profile was confirmed by ex vivo analysis of tumor extracts, and it was also observed in human surgical biopsies of IDH1-mutated tumors by 31P high resolution-magic angle spinning spectroscopy. The specificity of this profile for the IDH1-R132H mutation also was established by in vitro 31P-NMR of extracts of cells overexpressing IDH1 or IDH1-R132H. Overall, our results provide evidence that IDH1-R132H mutation alters phospholipid metabolism in gliomas involving phosphoethanolamine and glycerophosphocholine. This new non-invasive biomarker can assist in the identification of the mutation and in research towards novel treatments that target aberrant metabolism in IDH1-mutant glioma.