Forty-eight 27-wk-old White Leghorn hens were assigned randomly to four diets containing 0, 1.25, 2.5, or 5.0% conjugated linoleic acid (CLA). Hens were fed the CLA diets for 2 wk before eggs were collected for the study. Classes of egg yolk lipids were separated, and fatty acid concentrations in total lipid, triglyceride (TG), phosphatidylethanolamine (PE), and phosphatidylcholine (PC) were analyzed to determine the incorporation of dietary CLA isomers into different classes of egg yolk lipids. The amounts of CLA incorporated into lipid, PC, PE, and TG of egg yolk were proportional to the levels of CLA in the diet. However, more CLA was incorporated in TG than in PC and PE. The incorporation rates of different CLA isomers into different classes of lipids also were significantly different: cis-9, trans-11 and cis-10, trans-12 CLA were deposited more in TG, but cis-11, trans-13 CLA deposition in TG was significantly less. There were large differences in the concentrations of cis-8, trans-10 CLA in PC and PE. The inclusion of CLA into the diet influenced the metabolism of polyunsaturated fatty acids. The contents of 5,8,11,14-eicosatetraenoic, 9,12-octadecadienoic, and 9,12,15-octadecatrienoic acids were decreased as dietary CLA increased. Three isomers of hexadecadienoic acid were found in egg yolk lipids from hens fed 5% dietary CLA. The detection of hexadecadienoic acid isomers in lipid indicates that the utilization of CLA as an energy source after the first round of beta-oxidation may be less favorable than that of 9,12-octadecadienoic acid.