Absorption of calcium and magnesium endogenous to human milk, as well as calcium and magnesium added as an exogenous supplement to human milk, was determined in 9 very low birth wt infants. Human milk, intrinsically labeled with stable isotopic tracers of calcium and magnesium, was prepared by administering isotopic tracers intravenously to a lactating woman. Different isotopic tracers, which were representative of calcium and magnesium in the supplement (Enfamil Human Milk Fortifier, Mead Johnson Nutritional Div.), were added to the intrinsically labeled milk. The fortified milk, which was labeled with two calcium tracers and two magnesium tracers, was given orally to the test subjects in a single feeding. True absorption of calcium and magnesium was determined from differences between the doses of tracer ingested and the quantities of tracer excreted in the feces. Stable isotopic tracers were quantified by fast atom bombardment mass spectrometry. These results demonstrate that the fractional absorptions of calcium in the human milk and the added mineral supplement are 80 and 82%, respectively. A total of 89% magnesium endogenous to human milk and 86% of magnesium derived from the mineral supplement was absorbed by the VLBW infants.