Eight laboratories conducted a test for the estimation of the bioavailability of iron from 4 sources, using depleted male albino rats. Ferrous sulfate was used as the reference standard. Ferric orthophosphate was found to have a relative biological value of 11 (range 6-22), an old sample of hydrogen-reduced iron 27 (range 15-41), and ferric citrate 96 (range 75-125). Good results were obtained with a simplified basal diet prepared without ingredients that had previously contributed variable quantities of iron. There was no apparent advantage in using the change in hemoglobin during the repletion period instead of the final homoglobin value as the criterion of response to iron supplements. Several statistical treatments of the data yielded similar conclusions regarding relative biological values of the iron sources.