n-3 PUFA supplementation helps in the prevention or treatment of inflammatory diseases and CVD. However, many supplementations reported so far are either a combination of n-3 PUFA or used large daily amounts of n-3 PUFA dosages. The present study investigated the influence of increasing dose intake of DHA on the fatty acid composition of phospholipids in neutrophils and on their capability to produce leukotrienes (LT) B4 and B5in vitro. Twelve healthy volunteers were supplemented with increasing daily doses of DHA (200, 400, 800 and 1600 mg, each dose in TAG containing DHA as the only PUFA and for a 2-week period). At the end of each supplementation period, neutrophil fatty acid composition, and LTB4 and LTB5 production were determined by GC and liquid chromatography–tandem MS, respectively. The DHA/arachidonic acid ratio increased in a dose-dependent manner with respect to the increasing doses of DHA supplementation and was significantly different from baseline after supplementation with either 400, 800 or 1600 mg DHA. The LTB5/LTB4 ratio was significantly increased compared to baseline after supplementation with 800 and 1600 mg DHA. LTB5/LTB4 and DHA/arachidonic acid ratios were correlated (r 0·531, P < 0·0001). The present data suggest that both changes in neutrophil lipid composition and LT production occurred with daily supplementation with 800 and 1600 mg DHA. The clinical benefits associated with these doses of DHA in inflammatory diseases remain to be investigated.