The effects of a conjugated linoleic acid (CLA) mixture of single isomers (50:50, w/w, cis9,trans11:trans10,cis12) and the individual isomers on (a) the production of resting and calcium ionophore stimulated 1 4 C-eicosanoids and (b) the incorporation of 1 4 C-arachidonic acid (AA) into membrane phospholipids of human saphenous vein endothelial cells were investigated. The CLA mixture and the individual isomers were found to inhibit resting production of 1 4 C-prostaglandin F 2 a by 50, 43 and 40%, respectively. A dose dependent inhibition of stimulated 1 4 C-prostaglandins was observed with the CLA mixture (IC 5 0 100 μM). The cis9,trans11 and trans10,cis12 (50 μM) isomers individually inhibited the overall production of stimulated 1 4 C-prostaglandins (between 35 and 55% and 23 and 42%, respectively). When tested at a high concentration (100 μM), cis9,trans11 was found to inhibit eicosanoid production in contrast to trans10,cis12 that caused stimulation. The overall degree of 1 4 C-AA incorporation into membrane phospholipids of the CLA (mixture and individual isomers) treated cells was found to be lower than that of control cells and the cis9,trans11 isomer was found to increase the incorporation of 1 4 C-AA into phosphatidylcholine. Docosahexaenoic acid, eicosapentaenoic acid and linoleic acid did not alter the overall degree of incorporation of 1 4 C-AA. The results of this study suggest that both isomers inhibit eicosanoid production, and although trans10,cis12 exhibits pro-inflammatory activity at high concentrations, the CLA mixture maintains its beneficial anti-inflammatory action that contributes to its anti-carcinogenic and anti-atherogenic properties.