A 6-month crossover diet plan was employed to study the effects on human serum lipids of adding margarine or butter to otherwise self-selected diets that included two eggs daily. Two groups of subjects were studied: 51 free-living normocholesterolemic and 20 hypercholesterolemic (greater than 240 mg dl). Four-day diet records in each interval showed that subjects ate about 16% of total dietary fat as either butter or margarine. Blood samples taken every 6 weeks showed variable mean serum total cholesterol (SCHOL), high-density-lipoprotein cholesterol (HDL-C), and serum triglycerides (STG). The normocholesterolemic subjects who ate butter first had by 24 weeks mean SCHOL values equal to their entry values; those who ate margarine first had increased SCHOL values throughout the study. By the end of the study, the hypercholesterolemic subjects showed either no change or a slight decrease in both SCHOL and HDL-C values. Specific fatty acids were distributed differently in the serum fractions of triacylglycerol (TGFA), cholesteryl esters (CEFA), and phospholipids (PLFA). These distributions remained constant in both normocholesterolemic and hypercholesterolemic subjects regardless of the type and amount of fat consumed.