Beef contains functional fatty acids such as conjugated linoleic acid and long-chain fatty acids. This review summarizes results from studies comparing the fatty acid composition of beef from cattle fed either grass or grain-based feed. Since functional lipid components are contributed through dietary consumption of beef, the fatty acid composition is reported on mg/100 g of meat basis rather than on a percentage of total fat basis. Beef from grass-fed contains lesser total fat than that from grain-fed in all breeds of cattle. Reduced total fat content also influences the fatty acid composition of beef. A 100 g beef meat from grass-fed cattle contained 2,773 mg less total saturated fatty acids (SFA) than that from the same amount of grain-fed. Grass-fed also showed a more favorable SFA lipid profile containing less cholesterol-raising fatty acids (C12:0 to C16:0) but contained a lesser amount of cholesterol-lowering C18:0 than grain-fed beef. In terms of essential fatty acids, grass-fed beef showed greater levels of trans-vaccenic acid and long-chain n-3 polyunsaturated fatty acids (PUFA; EPA, DPA, DHA) than grain-fed beef. Grass-fed beef also contains an increased level of total n-3 PUFA which reduced the n-6 to n-3 ratio thus can offer more health benefits than grain-fed. The findings signify that grass-fed beef could exert protective effects against a number of diseases ranging from cancer to cardiovascular disease (CVD) as evidenced by the increased functional omega-3 PUFA and decreased undesirable SFA. Although grain-fed beef showed lesser EPA, DPA, and DHA, consumers should be aware that greater portions of grain-fed beef could also achieve a similar dietary intake of long-chain omega-3 fatty acids. Noteworthy, grain-fed beef contained higher total monounsaturated fatty acid that have beneficial roles in the amelioration of CVD risks than grass-fed beef. In Hanwoo beef, grain-fed showed higher EPA and DHA than grass-fed beef.