Volumetric analysis of hippocampal subfields and their asymmetry assessment recently has been useful biomarkers in neuroscience. In this study, hippocampal subfields atrophy and pattern of their asymmetry in the patient with Alzheimer's disease (AD) and mild cognitive impairment (MCI) were evaluated. MRI images of 20 AD patients, 20 MCI patients, and 20 healthy control (HC) were selected. The volumes of hippocampal subfields were extracted automatically using Freesurfer toolkit. The subfields asymmetry index (AI) and laterality ([Formula: see text]) were also evaluated. Analysis of covariance was used to compare the subfields volume between three patient groups (age and gender as covariates). We used ANOVA (P < 0.05) test for multiple comparisons with Bonferroni's post hoc correction method. Hippocampal subfields volume in AD patients were significantly lower than HC and MCI groups (P < 0.02); however, no significant difference was observed between MCI and HC groups. The asymmetry index (AI) in some subfields was significantly different between AD and MCI, as well as between AD and HC, while there was not any significant difference between MCI groups with HC. In all three patient groups, rightward laterality ([Formula: see text]) was seen in several subfields except subiculum, presubiculum, and parasubiculum, while in AD patient, rightward lateralization slightly decrease. Hippocampal subfields asymmetry can be used as a quantitative biomarker in neurocognitive disorders. In this study, it was observed that the asymmetry index of some subfields in AD is significantly different from MCI. In AD, patient rightward laterality was less MCI an HC group.