The changes in fatty acid composition of serum and in platelet aggregation induced by supplementation of arachidonate-enriched TAG were investigated in twenty-four healthy Japanese men in a double-blind, placebo-controlled study. The arachidonate-enriched TAG ingested was an edible oil, extracted and purified from a biomass of submerged fermented Mortierella alpina. Mean daily intake of fish and shellfish by subjects was 87.2 (se5.3) g/d, while dietary intakes of arachidonic acid (ARA) by the ARA group and placebo group were 175 (se12) and 179 (se13) mg/d, respectively. In the ARA group, after 2-week supplementation of 838 mg ARA/d, ARA concentration in serum phospholipids was increased from 9.6 (se0.4) to 13.7 (se0.4) g/100 g total fatty acids, and was significantly different from that in the placebo group (P < 0.001). This level was maintained for 4 weeks but returned to baseline level after a 4-week washout period. Linoleic acid concentration in serum phospholipids decreased from 19.2 (se0.8) to 16.3 (se0.6) g/100 g total fatty acids in the ARA group. Similarly, ARA content of serum TAG increased after ARA supplementation. Neither the EPA nor DHA content of serum phospholipids or TAG was altered by ARA supplementation. The platelet aggregation induced in platelet-rich plasma by adding adenosine diphosphate, collagen and ARA, physical characteristics of subjects, and biochemical parameters were unchanged throughout the test period. These findings suggest that ARA concentration in serum phospholipids and TAG can be safely increased by supplementation of arachidonate-enriched TAG oil.