Factors associated with concentrations of energy-yielding nutrients in human milk were examined at 3, 6, 9, and 12 mo postpartum in the DARLING (Davis Area Research on Lactation, Infant Nutrition and Growth) Study. Samples were obtained by complete expression of alternate breasts over 24 h. Milk energy density was highly correlated with lipid concentration; both were positively related to maternal percent of ideal body weight (%IBW) at 6, 9, and 12 mo and negatively related to milk volume at 3 mo and to parity at 12 mo. Milk protein concentration was negatively related to milk volume at 6 and 9 mo and positively related to nursing frequency at 6 mo and %IBW at 9 mo. Milk lactose concentration was positively related to milk volume at 6 and 9 mo and to continued amenorrhea at 9 mo. In a subsample who completed dietary records, protein intake was positively associated with lipid concentration after 16 wk postpartum but not before. These findings suggest that milk composition is more sensitive to maternal factors such as body composition, diet, and parity during later lactation than during the first few months.