High-resolution 1H NMR spectroscopy has been used to measure the concentrations of metabolites (alanine, N-acetylaspartate, gamma-aminobutyric acid, glutamate, glutamine, aspartate, taurine, glycine, succinate, creatine, cholines, inositol, and glucose) in perchloric acid extracts of human epileptic cortex and brain tumors. All tissue was obtained by surgical biopsy, excised before thermal coagulation, and immediately frozen in liquid nitrogen. Lower levels of N-acetylaspartate and gamma-aminobutyric acid and a shift in the glutamate/glutamine ratio toward glutamine in the tumors reflect neuronal loss. Abnormal glucose metabolism (aerobic glycolysis) in the tumors gives decreased levels of succinate, glutamate, aspartate, glutamine, and creatine and generally increased concentrations of glycine and alanine. Differences in metabolite concentrations that may be of use in differential tumor diagnosis include lower creatine and inositol in meningiomas than in astrocytomas. Lower taurine differentiates benign from malignant astrocytomas. Malignant astrocytomas and metastatic tumors are more regionally heterogeneous than meningiomas or benign astrocytomas. Mannitol, administered perioperatively to all patients from whom tissue was obtained, was observed only in the spectra of extracts of tissue from tumors which enhanced on computerized tomographic imaging.