Recent studies have demonstrated that long-chain n-3 PUFA (LCn-3PUFA) are beneficial in reducing the risk of cardiac arrhythmias. This study was conducted to determine the extent of incorporation of LCn-3PUFA into human atrium following supplementation with a fish oil concentrate high in LCn-3PUFA. Volunteers preparing for coronary bypass surgery were randomized either to the treatment group (n = 8), receiving 6 g/d of fish oil concentrate (4.4 g of LCn-3PUFA), or the placebo group (n = 9), receiving 6 g/d of olive oil for a minimum period of 6 wk. Blood samples were collected prior to commencement of treatment, and preoperatively before bypass surgery. Atrial biopsies were obtained during surgery. The plasma and atrium samples were analyzed by GC following trans-methylation to determine FA profile. Post-supplementation, the treatment group had significantly higher plasma levels of 20:5n-3, 22:5n-3, and 22:6n-3 than the placebo group. Analysis of the atrium total lipids revealed a significant increase in the proportion of 20:5n-3 following fish oil supplementation. There was no significant difference in the concentration of 22:5n-3 and 22:6n-3 in the atrium total lipids; however, an upward trend was observed in subjects receiving fish oil supplementation. In the phospholipid fraction of the atrium, both 20:5n-3 and 22:6n-3 increased, whereas 20:4n-6 levels decreased. This study demonstrates for the first time that short-term supplementation with fish oil concentrate results in significant incorporation of LCn-3PUFA with a concomitant depletion of the eicosanoid substrate (20:4n-6) in the human atrium.