The effects of zero-erucic acid rapeseed oil and rapeseed oil-containing margarine on plasma fatty acid composition and serum cholesterol were studied in butter users (n 43). Compliance to the substitution was followed by fatty acid analysis of total plasma and plasma phospholipids. The amount of substitute fats represented, on average, 21% of total fat and 8% of total energy intake. Changes in the relative fatty acid composition of plasma phospholipids indicated further fatty acid metabolism, and were closely related to the serum cholesterol level. The reduction in saturated fatty acids led to a significant increase in the proportion of n-3 and n-6 polyunsaturated fatty acids (PUFA) with the rapeseed oil diet, whereas the margarine caused a significant rise in n-6 PUFA only. The increase in the proportions of the two PUFA families occurred in accordance with their competitive order, most completely with the rapeseed oil diet. When butter was replaced by rapeseed oil, low-density-lipoprotein-cholesterol decreased by an average of 9.1% without a reduction in high-density-lipoprotein-cholesterol. During margarine substitution the reduction was 5.2%, on average. Of the plasma phospholipids, alpha-linolenic acid and the linoleic:stearic acid ratio, but not oleic acid, were the components most significantly correlated with serum cholesterol levels or the decrease in these levels. The results show that rapeseed oil can act primarily as a source of essential fatty acids, rather than that of monoenes, in the diet of butter users.