This study characterizes B vitamin supplement use prior to and during pregnancy, changes in dietary one-carbon nutrient intakes (folate, vitamin B12, vitamin B6, choline, betaine and methionine) and most significant dietary sources. In Canadian women (Toronto, Ontario) supplemental (n=364) and dietary intakes (using a food frequency questionnaire) (n=290) were assessed during early and late pregnancy. Majority reported using a B vitamin-containing supplement prior (60%) to and during early (93%) and late (89%) pregnancy. Median supplemental intakes of folic acid, B12 and B6 were 1000 µg/d, 2.6 µg/d and 1.9 mg/d, respectively. Dietary one-carbon nutrient intakes did not change appreciably between early and late pregnancy. Most significant sources of folate and B6 were fruits and vegetables, of folic acid were cereals and grains and of B12 were dairy and egg products. Overall, this study provides novel information about one-carbon nutrient intakes in pregnancy which are crucial in maternal and child health.MAS