Ten healthy volunteers were given a daily supplement of 30 g olive oil for 6 weeks in order to evaluate how it would affect cell membrane composition and ultimately platelet function. Fasting blood and cheek cell samples were taken before commencing the study, after 21 and 42 days of supplementation and also at 30 days after finishing the supplement (washout). C18:1n-9 was significantly increased (p < 0.01) in platelet and cheek cell phospholipids. Erythrocytes were not good markers for C18:1n-9 intake and no significant change was found in this tissue. There was a small nonsignificant decrease in platelet phospholipid 20:4n-6 after the supplementation, in accordance with previously published results. C18:1n-9 did not persist in platelet membranes after the volunteers stopped consuming the olive oil supplement, but in erythrocytes a significant increase (p < 0.05) was found after the washout period. None of these changes in fatty acid composition in the different tissues were related to significant changes in serum cholesterol-related variables or in clotting factors or adenosine diphosphate-induced platelet aggregation.