This study investigated the effects of a moderate dose of long-chain n-3 polyunsaturated fatty acids (1.8 g eicosapentaenoic acid (EPA) plus 0.3g docosahexaenoic acid (DHA) per day) given for 8 weeks to healthy middle-aged males on cardiovascular risk factors, particularly plasma lipids and inflammatory markers. The study was double-blind and placebo-controlled. The proportion of EPA was significantly increased in plasma phosphatidylcholine (from 1.4% to 5.0% of total fatty acids; P<0.001), cholesteryl esters (from 1.2% to 4.5%; P<0.001) and triacylglycerols (from 0.3% to 1.8%; P<0.001). In contrast, the more modest increases in DHA in these lipid fractions were not significant. There was very little effect of n-3 fatty acids on the risk factors measured, apart from a reduction in plasma soluble intercellular adhesion molecule (sICAM)-1 concentration compared with placebo (P=0.05). The change in plasma sICAM-1 concentration was significantly inversely related to the change in DHA in plasma phosphatidylcholine (r=-0.675; P=0.001), but less so to the change in EPA (r=-0.406; P=0.076). Data from the present study suggest that marine oil providing 1.8 g of EPA plus 0.3g DHA/day is not sufficient to demonstrate marked effects on cardiovascular risk factors (plasma lipids and inflammatory markers) in healthy middle-aged men, although there may be a slight anti-inflammatory effect as indicated by the decrease in sICAM-1. The stronger association between changes in DHA than EPA and sICAM-1 concentrations suggest that DHA may be more anti-inflammatory than EPA. Thus, one reason why only limited effects were seen here may be that the dose of DHA provided was insufficient.