Milk triglycerides from the platypus were subjected to fatty acid and stereospecific analysis to determine the positional distribution of fatty acids in the triglycerides. Of the major fatty acids, 12∶0 was preferentially esterified at thesn-3 position, 14∶0 and 16∶0 were selectively associated with thesn-2 position, and 18∶0 was located predominantly at thesn-1 position. The unsaturated fatty acids, 14∶1, 16∶1, 18∶1, 18∶2 and 18∶3, were preferentially esterified at thesn-3 position. The fatty acid distribution pattern of the platypus, a monotreme, is similar to that of marsupials and eutherians but is in contrast to the only other extant monotreme, the echidna.