BACKGROUND The nutritional effect of the regiospecific distribution of fatty acids in edible fats is currently discussed due to an increased use of interesterification of fats for human consumption. However, disagreeing results have been reported which may be due to the varying composition of the dietary fats compared. Data on the fate of such lipids beyond the bloodstream is rather scarce and animal model studies are needed. OBJECTIVE To compare the metabolism of butter oil and mixtures of butter and rapeseed oil, native or randomized, in a rat model. The regiospecific fatty acid distribution present in dietary fats was followed through absorption, chylomicron formation, and deposition in adipose tissue and in different liver lipids (triacylglycerols, phosholipids, and cholesterol esters). METHODS Rats were fed for 6 weeks from weaning either butter oil (BO), a butteroil-rapeseed oil mixture 65:35 w/w (BR) or a randomized mixture of BR (tBR). Half of the animals were used for organ analysis, the rest for a postprandial study with the same fats and isolation of chylomicrons. The regiospecific distribution of the fatty acids present in the dietary fats was followed during metabolism by analyses of chylomicrons, depot fat and liver lipids, using regiospecific cleavage followed by TLC separation and quantification by GC. RESULTS Randomization of edible fat mixtures leading to equal distribution of fatty acids between TG positions is directly reflected in the composition of chylomicrons. During clearing by lipoprotein lipase this positional distribution is abolished and the regiospecific composition of triacylglycerols in adipose tissue is completely identical for BR and tBR. Chylomicron remnants, which are taken up by the liver, are correspondingly fully degraded to free fatty acids by hepatic lipase, and distribution of fatty acids in liver triacylglycerols, phospholipids and cholesterol esters are identical for the groups fed either BR or tBR. The group fed BO with a low content of linoleic acid is on the borderline of essential fatty acid-deficiency. CONCLUSIONS Randomization (interesterification) of butter oil with rapeseed oil (65:35 w/w) for use as edible fat did not have any impact on the fatty acid composition beyond the chylomicron step when compared to the native mixture.