The fatty acid composition of total lipids, triglycerides, free fatty acid pool, esterified cholesterol, phosphatidyl choline, and phosphatidyl ethanolamine of mammary tissue of lactating and nonlactating buffalo has been studied. The distribution of long chain triglycerides, free fatty acids free cholesterol and esterified cholesterol in nonlactating mammary tissue was 76.1,4.2,4 and 0.5 percent of total lipids respectively and in lactating mammary tissue it was 68.3, 1.2, 2.7 and 0.3 percent respectively. Short chain triglycerides constituting about 6.6% of total lipids and could be detected only in lactating mammary tissue. Among phospholipids, phosphatidyl choline, phosphatidyl ethanolamine and sphingomyelin constituted about 39.4, 21.5 and 15.6 percent respectively of total phospholipids in nonlactating tissue and 39.3, 22.6 and 10.7 percent respectively in lactating tissue. All the major fatty acids viz myristic, palmitic, stearic acid oleic were present in various lipid components of both lactating and nonlactating mammary tissue. Linoleic acid was present only in the lactating mammary tissue. Differences were observed in the distribution of fatty acids in lipid components of lactating and nonlactating mammary tissue.