The aim of this study was to evaluate the fatty acid composition of intramuscular fat and the vitamin E content of beef from different production systems. Four cattle production systems were used: cattle reared under intensive conditions and fed concentrate (INT1) from Spain, cattle raised at pasture and supplemented with concentrate (SUP1) from the United Kingdom, cattle raised at pasture and on corn silage (SUP2) from Germany and cattle reared under extensive conditions slaughtered at two and three years old (EXT2 and EXT3) from Uruguay. The highest proportion and content (mg per 100g of muscle) of linoleic acid (C18:2n-6) and arachidonic acid (C20:4n-6) and the lowest saturated fatty acids (SFA) was found in INT1 beef. In contrast, beef reared under extensive conditions showed the highest proportion and content of linolenic acid (C18:3n-3), docosapentaenoic acid (DPA, C22:5n-3) and eicosapentaenoic acid (EPA, C20:5n-3), and SUP1 and SUP2 beef showed the highest level of palmitic acid (C16:0) and SFA. Beef from intensive system (INT1) had the lowest P/S (PUFA/SFA) ratio, whereas beef from extensive system (EXT2 and EXT3) had the lowest n-6/n-3 ratio. The results of the PCA (principal components analysis) of fatty acid composition showed that beef from intensive system (INT1) was clearly differentiated from the other meats and was located with the C18:2n-6 and C20:4n-6 and the n-6/n-3 ratio. EXT2 and EXT3 beef were located with C18:3n-3 and long chain fatty acids. Beef from extensive systems had the highest concentration of vitamin E (3.91mg α-tocopherol/kg muscle).