We evaluated the effect of moderate dietary changes on the prostaglandin system by measuring the urinary excretion of 7 alpha-hydroxy-5,11-dioxo-tetranorprostane-1,16-dioic acid (PGE-M). In a crossover design, twenty-four free-living male subjects in good health (24 to 54 years of age) were fed two diets: (i) Regular (R) diet, 41% energy (en%) from fat, P/S 0.59, M/S 0.96; (ii) Experimental (E) diet, 19 en% from fat, P/S 1.31, M/S 1.48. Diet R contained 13.9 g/day of neutral detergent fiber (NDF) and about 600 mg/day of cholesterol per 3200 kcal; Diet E contained 35.5 g/day NDF and about 280 mg/day cholesterol. Each controlled-diet period lasted ten weeks. The menu cycle was 7 days, and all diets were calculated to provide adequate amounts of essential nutrients. The PGE-M excretion rates were determined in 24-hr urine by stable-isotope dilution gas chromatography-mass spectrometry in the selected ion-monitoring mode. Low-fat Diet E, with an intake of 6.6 en% from polyunsaturates, was associated with an average 14.2% reduction in PGE-M daily output, compared to high-fat Diet R with a 9.3 en% from polyunsaturates (P = 0.046). These results support the view that dietary lipid changes can significantly alter the in vivo production of E-series prostaglandins. We cannot conclude, however, if this apparent diet effect was brought about by the change in linoleate intake alone or was the result of complex biochemical interactions among individual fatty acids, both saturated and unsaturated.