Collaborative Study of the Rat Hemoglobin Repletion Test for Bioavailability of Iron

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Abstract
A collaborative study of the hemoglobin repletion test was conducted with male weanling rats. The rats were depleted for 4 weeks on a low-iron casein based diet. They were then divided into 13 comparable groups of at least 8 animals each. One group continued on the basal diet. Three groups were given different levels of each of 4 samples. Sample 1 was FeS04.7H20, used as the reference standard. Samples 2 and 3 were from the same lot of electrolytically reduced iron separated into fractions of different particle size by nitrogen elutriation, and were 7—10 and 27—40 /tm, respectively. Sample 4 was a mixture of several lots of food grade ferric orthophosphate. After 2 weeks on the test diets, individual blood samples were drawn for hemoglobin determinations. The data were analyzed by the parallel lines technique, and relative biological values, vs. FeS04 = 100, together with 95% confidence limits were calculated. The relative biological values were as follows: reduced iron, 7—10 /mi = 63.5 ± 10.9; reduced iron, 27— 40 /im = 3 7 . 9 ± 1 2 . 2 ; ferric orthophosphate = 44.5±4.8. This revised method for measuring bioavailability of iron has been adopted as official first action to replace the original method, 39.A15-39.A17.

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