The goal of the present study was to investigate inter‐individual and age‐dependent variation of global DNA methylation in human tissues. In this work, we examined 5‐methyldeoxycytidine (^metC) content by HPLC in human peripheral blood leukocytes obtained from 76 healthy individuals of ages varying from 4 to 94 years (yr), and 39 human placentas from various gestational stages. The HPLC analysis revealed a significant variation of ^metC across individuals and is consistent with the previous findings of age‐dependent decrease of global methylation levels in human tissues. The age‐dependent decrease of ^metC was relatively small, but statistically highly significant (p= 0.0002) in the aged group (65.9 ± 8.9 [mean age ± SD] yr; n = 22) in comparison to the young adult group (19.3 ± 1.4 yr; n = 21). Males showed a subtle but statistically significant higher mean ^metC content than females. In contrast to the peripheral blood samples, DNA extracted from placentas exhibited gestational stage‐dependent increase of methylation levels that appeared to inversely correlate with the expression levels of human endogenous retroviruses. These data may be helpful in further studies of DNA methylation, such as inheritance of epigenetic patterns, environment‐induced changes, and involvement of epigenetic changes in disease.