Glutamate (Glu) is the most abundant excitatory neurotransmitter in the central nervous system (CNS) and is involved in the pathophysiology of Alzheimer's disease (AD) in which there is an increased excitotoxicity. Biochemical composition of living tissues including the levels of Glu was analyzed by magnetic resonance spectroscopy (MRS). Previous reports point to decreased levels of Glu in AD. As Glu plays an important role in memory, we hypothesize that Glu levels are decreased in patients with AD when compared with controls. A consecutive sample of 30 patients with mild-to-moderate AD underwent H-MRS with the voxel placed in the bilateral posterior cingulate gyrus. For comparison purposes, we carried out the same technique in 68 patients with mild cognitive impairment (MCI) and in 26 controls. The healthy controls had higher metabolite levels of N-acetyl-aspartate (NAA) than patients with MCI and AD. In turn, patients with MCI and the controls had higher levels of Glu than in patients with AD. The differences were significant in the analysis of variance (ANOVA) test model corrected for age. In the post hoc analysis, the most remarkable differences were seen between patients with AD and the rest (patients with MCI and the controls). In AD, the levels of Glu and NAA are decreased in comparison with MCI and normality, which reflects loss of neurons.