The calcium requirement for prolonged lactation was investigated in a randomized supplementation study of Gambian mothers consuming a low-calcium diet (7.1 mmol/d, or 283 mg/d). Sixty women were studied from 10 d to 78 wk of lactation, receiving calcium or placebo for the first 12 mo. The supplement increased average calcium intake by 17.9 mmol/d (714 mg/d). Supplementation had no effect on breast-milk calcium concentration or on maternal bone mineral content. Urinary calcium output was higher in supplemented than in unsupplemented mothers by 1.18 mmol/d (47 mg/d), P < or = 0.005. Longitudinal changes in urinary calcium output and bone mineral content made a substantial contribution to calcium requirements for lactation. This study suggests that, in women with low calcium intakes, there is no direct benefit from increasing calcium intake during lactation, and that physiological mechanisms operate to furnish calcium for breast-milk production.