Milk was collected from mothers of 18 very premature (26 to 30 wk gestation age), 28 premature (31 to 36 wk), and six term (37 + wk) infants on day 2 to 3 (colostrum), and at 1, 3, 6, and 12 wk postpartum. Fat content for 154 milk samples was 2.80 g/dl gravimetrically and 2.66 g/dl by quantitative thin-layer chromatography. Fat content increased during lactation, whereas phospholipids and cholesterol declined. Concentrations of medium-chain fatty acids increased from colostrum to mature milk and were highest in preterm milk. Compensatory decreases were observed in very premature and premature oleic acid. Long-chain polyunsaturated fatty acids were highest in colostrum and reduced in mature milk. Long-chain polyunsaturated fatty acids were also higher in very premature and premature milk than in term milk. These elevated levels of readily absorbed medium-chain fatty acids and long-chain polyunsaturated fatty acids in preterm milk may be of special benefit for the needs of premature infants.