The selective incorporation of dietary α-linolenate (18:3n-3) and its elongation product, eicosatrienoate (20:3n-3), into various phospholipids (PL) of mouse liver, spleen, kidney, and heart, was examined in a two-week feeding trial by assessing mol % changes in associated fatty acids. Mice were fed fat-free AIN 76A diets modified with either 2 wt% safflower oil (control); 1% safflower and 1% linolenate; or 1% safflower and 1% eicosatrienoate. After linolenate or eicosatrienoate feeding, 20:4n-6 was reduced by 36-50% in liver phosphatidylcholine (PC) and in liver and spleen phosphatidylethanolamine (PE). Linolenate was minimally incorporated into PL, but was desaturated and elongated to 20:5n-3, 22:5n-3, and 22:6n-3, with notable differences in the quantity of these n-3 derivatives associated with different tissues and PL. Eicosatrientoate was uniquely incorporated into the cardiolipin (CL) pool of all organs. There was also considerable retroconversion of 20:3n-3 to 18:3n-3 (PC, PE). Dietary eicosatrienoate may therefore affect metabolism in diverse ways-20:3n-3, which is retroconverted to 18:3n-3, may provide substrate for 20:5n-3 and 22:6n-3 syntheses, whereas intact 20:3n-3 may be incorporated into the CL pool. Acyl modifications of CL are known to affect the activity of key innermitochondrial enzymes, such as cytochrome c oxidase. © 1990 American Oil Chemists' Society (AOCS).