We investigated the inhibitory effect of calcium on iron absorption in 57 human subjects. Three studies suggested that the effect is not located in the gastrointestinal tract. The presence of phytate in a meal and formation of calcium-iron-phytate complexes is not a prerequisite for the inhibition. The relative increase in iron absorption by ascorbic acid was the same in meals with and without calcium, suggesting that calcium did not influence the balance between enhancing and inhibiting ligands in the gastrointestinal lumen. No inhibiting effect on iron absorption was seen when adding 3 mg calcium to 0.01 mg iron (molar ratio Ca/Fe = 420). Previous studies showing a marked inhibition by calcium had a lower molar ratio, but greater amounts of calcium were given. This suggests that a minimal concentration of calcium is needed to achieve an effect. The present results indirectly support our original hypothesis that the inhibitory effect of calcium on iron absorption is situated within the intestinal mucosal cells. The practical nutritional implications of the inhibitory effect of calcium are considerable since addition of milk, milkshake or cheese to common meals such as pizza or hamburger meals reduced iron absorption by 50-60%. It is recommended to reduce the intake of dairy products with the main meals providing most of the dietary iron, especially for those having the highest iron requirements i.e. children, teenagers and women at childbearing age.