Background/Aims: An important public health issue is monitoring folate inadequacy in women of childbearing potential. The aim of the present study was to investigate associations between folate intake, red blood cell (RBC) folate, total homocysteine (tHcy) and the MTHFR 677T allele. Methods: A total of 204 women were enrolled in a cross-sectional study. Folate intake was assessed by a food frequency questionnaire, and RBC folate, tHcy and MTHFR C677T genotype were determined. Results: About half of the women had a decreased RBC folate level (<305 nmol/l) and all were <906 nmol/l, even though 51% of the subjects reported use of supplements. Overall 91.5% had a high Hcy concentration. Notably, younger women, and those with a low level of education, were shown to be at higher risk of inadequate RBC folate levels. Additionally, younger women were also at higher risk of carrying the TT genotype, particularly unfavorable in the setting of a low folate status. Conclusions: Our study revealed significant folate deficiency in our Mediterranean population and higher than ideal Hcy concentrations, thus emphasizing that in these groups an improvement in the folate status is needed via a food-based approach or supplement. Consequently, public health policy strategies aiming at improved supplementation are required.