Low red blood cell (RBC) membrane content of EPA + DHA (hereafter called omega-3 index) has recently been described as an indicator for increased risk of death from coronary heart disease. The relationship between plasma and RBC FA, focusing on omega-3 index, and the response to DHA supplementation were investigated in a double-blind, randomized, placebo-controlled, intervention study. Healthy vegetarians (87 f, 17 m) consumed daily a microalgae oil from Ulkenia sp. (0.94 g DHA/d) or olive oil (placebo) for 8 wk. DHA supplementation significantly increased DHA in RBC total lipids (7.9 vs. 4.4 wt%), in RBC PE (12.1 vs. 6.5 wt%), in RBC PC (3.8 vs. 1.4 wt%), and in plasma phospholipids (PL) (7.4 vs. 2.8 wt%), whereas EPA levels rose to a much lesser extent. Microalgae oil supplementation increased the omega-3 index from 4.8 to 8.4 wt%. After intervention, 69% of DHA-supplemented subjects (but no subject of the placebo group) reached an omega-3 index above the desirable value of 8 wt%. Omega-3 index and EPA + DHA levels in RBC PE, RBC PC, and plasma PL were closely correlated (r always > 0.9). We conclude that an 8-wk supplementation with 0.94 g DHA/d from microalgae oil achieves a beneficial omega-3 index of > or =8% in most subjects with low basal EPA + DHA status. RBC total FA analyses can be used instead of RBC lipid fraction analyses for assessing essential FA status, e.g., in clinical studies.