We investigated the effect of fish oil supplementation on the synthesis of prostaglandin E (PGE) in vivo by measuring the excretion of its catabolite, PGE-M, in 24-hr urine by gas chromatography/mass spectrometry. Forty healthy male volunteers (24-57 years of age) consumed a controlled basal diet providing 40% of energy from fat (P/S ratio about 0.8:1), 130 mg/1000 kcal cholesterol, and a minimum of 22 mg/day of alpha-tocopherol (alpha-T), for three experimental periods lasting a total of 28 weeks. During period 1 (10 weeks) the diet was supplemented with placebo (PO) capsules (15 X 1 g/day) consisting of a blend of fats approaching the fatty acid profile of the basal diet. This was followed by a second 10-week period during which the subjects received 15 X 1 g/day capsules of fish oil concentrate (FOC). During period 3 (8 weeks) they continued the 15 g/day intake of FOC but received an additional 200 mg/day of alpha-T. PO and FOC capsules contained 1 mg alpha-T/g fat as antioxidant. A 14% reduction of PGE-M excretion was observed after 10 weeks of FOC supplementation (period 2), compared to an identical period of placebo supplementation (period 1), P = 0.009. PGE-M excretion during the last week of period 3 was not significantly different from that at the end of period 2. The reduction in PGE synthesis in response to the relatively high marine oil supplementation was large in many subjects participating in this study.