The influence of maternal intake of vitamin C on the vitamin C concentration in human milk and on the vitamin C intakes of breast-fed infants has not been demonstrated conclusively. This study examined these influences of diet and supplementation in 25 lactating women administered 90 mg of ascorbic acid for 1 day followed by 250, 500 or 1000 mg/day for 2 days or unsupplemented for 1 day followed by either 0 or 90 mg ascorbic acid supplement for 2 days. Vitamin C content in milk and urine was determined by the 2,4-dinitrophenylhydrazine method. Vitamin C intakes of infants were calculated from milk volume, as determined by the test-weighing method and from vitamin C levels in milk samples obtained at each feeding. Total maternal intakes of vitamin C, which exceeded 1000 mg/day or 10-fold the RDA for lactation (100 mg/day), did not significantly influence the vitamin C content in milk or the vitamin C intakes of infants. However, maternal vitamin C intake was positively correlated (r = 0.7) with maternal urinary excretion. These differences in milk and urine response to vitamin C intake suggest a regulatory mechanism for vitamin C levels in milk.