Prior investigations have shown that rats are less sensitive than humans to dietary factors that influence the absorption of nonheme iron. This investigation was undertaken to determine whether this disparity is due to differences in the methods used to measure absorption in the two species. By use of identical methodology and test meals, absorption studies were performed in rats and humans to compare the effect of known dietary enhancers (ascorbic acid and meat) and inhibitors (tea, bran, and soy protein) on nonheme-iron absorption. Meat and tea had a marked effect on absorption in humans but did not influence absorption in rats. Although the effect of ascorbic acid, soy protein, and bran on absorption was statistically significant in rats, the absorptive response was far less than it was in humans. Our studies indicate that rodents cannot be used to assess the quantitative importance of dietary factors in human iron nutrition.