Objective: To determine the effect of folic acid, vitamin B6 and B12 fortified spreads on the blood concentrations of these vitamins and homocysteine. Design and setting: A 6-week randomized, double-blinded, placebo-controlled, parallel trial carried out in a clinical research center. Subjects: One hundred and fifty healthy volunteers (50% males). Interventions: For 6 weeks, the subjects consumed the test spreads (20 g/day): containing per 20 g (1) 200μ g folic acid, 2μ g vitamin B12 and 1 mg vitamin B6, or (2) 400μ g folic acid, 2μ g vitamin B12 and 1 mg vitamin B6 or (3) no B-vitamins (control spread). Results: The B-vitamin status increased on using the test spreads, with the largest effect on the serum folate concentration: 48% in men and 58% in women on spread 1 and 92 and 146%, respectively, on spread 2 (P-values all <0.05). The plasma homocysteine decreased in the groups treated with the fortified spreads as compared to the control group. Average decreases were for males: 0.7±1.5±μmol/l (6.8%) on spread 1 and 1.7 ± 1.7 μmol/l (17.6%) on spread 2 and for females: 1.4 ± 1.2 μmol/l (14.2%) and 2.4 ± 2.0 μmol/l (23.3%), respectively (P-values all <0.05). Conclusions: Consumption of a spread fortified with folic acid, vitamin B6 and vitamin B12 for 6 weeks significantly increases the blood concentrations of these vitamins and significantly decreases the plasma concentration of homocysteine. Fortified staple foods like spreads can contribute to the lowering of homocysteine concentrations.