A stable isotope technique has been developed which uses 57Fe and 58Fe as labels and which enables the simultaneous measurement of Fe absorption from two test meals in infants. The method was evaluated by measuring Fe absorption from a commercial whey-adjusted infant formula in nine healthy infants aged 13-25 weeks. Each infant was fed 210 ml formula, labelled with either 57Fe or 58Fe, on four consecutive mornings, in random order. The total Fe content in each feed was 2.5 mg Fe; either as 2.5 mg 57Fe, or 0.6 mg 58Fe plus 1.9 mg Fe with normal isotopic composition. Isotopic enrichment of Fe in erythrocytes was measured by thermal ionization mass spectrometry 14 d after the last administration, and Fe absorption was calculated based on isotope ratio shifts, total circulating Fe and intake of each isotope. Geometric mean absorption for the 57Fe and 58Fe labels was 6.72 and 6.58% respectively, and the absorption of the two isotopes was not significantly different (Student's paired t test). By this technique, paired comparisons of Fe absorption can be obtained and systematic studies of the influence of dietary factors on Fe absorption during infancy can be conducted.