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A study was undertaken to define an appropriate marker of lactogenesis II (the onset of copious milk secretion) in mothers, and to determine the effect of diabetes on this marker. Changes in the concentrations of three milk components—lactose, citrate, and glucose—were measured in 38 normal mothers and 6 type I diabetic mothers up to 10 days after birth. Milk yield was measured in 12 of the normal mothers, and all mothers were asked to note the time of milk “coming in” (the feeling of overfullness of the breasts). The average concentrations of lactose, citrate, and glucose in milk were low for the first 24 h after birth, then between 24 and 48 h after birth there was a rapid increase in the concentrations of lactose and citrate, and this transitional period was followed by a plateau period that began between 60 and 84 h after birth. For individual mothers the transitional period for citrate began 32 ± 9 h (n = 13) and finished 77 ± 10 h (n = 17) after birth, and for lactose the transitional period finished at 53 ± 12 h (n = 29) after birth. For diabetic mothers these times were significantly later. The average 24-h milk intake by infants increased from 82 to 556 ml/24 h between 24 and 144 h after birth. Milk intakes were correlated with the concentration of lactose (r = 0.52, n = 51, p < 0.001), citrate (r = 0.47, n = 47, p < 0.001), and glucose (r = 0.69, n = 50, p < 0.001). The time of milk “coming in” was highly variable (59 ± 13) (X ± SD) and occurred well after the onset of copious milk secretion between 24 and 48 h after birth. We conclude that for individual mothers, milk lactose and citrate are useful markers of lactogenesis II. The changes in the concentrations of these markers suggested that lactogenesis II was delayed in type I diabetic mothers.