Distribution of intestinal iron and absorption were studied in rats with high hemoglobin (Hb) levels of >7.5 g/dL or low-Hb, ≤7.5 g/dL, which were fasted overnight and fed a single 2-gram meal of diet containing either wheat bran or FeSO4. One hour later they were gavaged with 2 μCi of 59FeCl3 in 0.5M HCl. Low-Hb rats absorbed similar amounts of iron from the bran and FeSO4 diets. Both groups of rats absorbed similar amounts of iron from FeSO4, but the high-Hb rats absorbed less iron from bran than did the low-Hb ones. Gastric 59Fe clearance was slower in the high-Hb than in the low-Hb rats. Gastric 59Fe clearance was faster from FeSO4 diet, regardless of iron status. About 10–20% of the 59Fe was in the ileum and about 1.3–3.3% of the 59Fe was in the colon after gavaging; the distal intestines of the rats fed the bran diet contained the least 59Fe. During the first 24 h, whole blood 59Fe activity was similar for both diets; but after 24 h, 59Fe activity was lower in the rats fed the bran diet. During the first 48 h, plasma clearance of i.v. injected 55Fe transferrin was greater in the low-Hb rats than in the high-Hb ones but plasma clearance was not affected by iron source. However, beginning at 36 h, clearance was reduced in rats fed the bran diet. Thus, the high-Hb rats (but not the low-Hb rats) differed in the mechanisms of their absorption of wheat bran and FeSO4 iron.