The most useful and appropriate methods for assessing the bioavailability of (nonheme) iron supplements are described. When the supplement can be labeled isotopically, the best method for measuring bioavailability is hemoglobin incorporation, followed by fecal monitoring. Caco-2 cell in vitro systems can be used for rapid screening to predict potential availability for absorption. If the compound cannot be labeled, then the plasma appearance/disappearance of oral iron given together with an intravenous dose of iron isotope can be used to quantify absorption. With oral doses in excess of 25 mg, the 4- to 6-h plasma concentration can provide a qualitative assessment of bioavailability. Approaches for normalizing results to minimize intraindividual and interindividual variability in efficiency of iron absorption are discussed.