The net rates of uptake of the natural (2 R,4′R,8′R) diastereoisomer of α-tocopherol (α-T) and the biodiscrimination relative to its 2 S-epimer (2 S,4′R,8′R) have been measured, in two experiments, for the blood and 21 tissues of male Sprague-Dawley rats fed over a period of several months diets containing deuterium-substituted forms of the α-T acetates. Gas chromatography-mass spectrometry was used to measure the amount of deuterated tocopherols taken up relative to the amount of nondeuterated tocopherol remaining. The measurements were performed at different times after the rats, placed for one month on a basal diet containing nondeuterated, natural α-T acetate, were switched to a diet containing the same total quantity of deuterated forms of either natural α-T acetate or a mixture of the acetates of the 2 R- and 2 S-epimers (i.e., ambo-α-T acetate). In experiment 1 the source of vitamin E in the replacement diet was trideuterio-2 R,4′R,8′R-α-T acetate. The data obtained provide the first direct measure of the rate at which natural vitamin E is replaced and augmented in the tissues of growing animals under normal laboratory dietary conditions. There are dramatic differences in the tissue kinetics; for example, the apparent half-life of vitamin E, i.e., the time at which the total amount of ingested trideuterio-α-T taken up is the same as the amount of nondeuterated α-T remaining, varies from ca. 1 wk for the lung to ca. 11 wk for the spinal cord. In experiment 2 the vitamin E in the replacement diet was an equimolar mixture of trideuterio-2 S,4′R,8′R- and hexadeuterio-2 R,4′R,8′R-α-T acetates. The results show that there is a preferential uptake of the natural diastereoisomer of α-T by all tissues (except the liver during the first month). Examination of fecal material reveals that the biodiscrimination begins in the gut; the incomplete hydrolysis of the acetates shows clearly that this reaction proceeds to a greater extent with the natural diastereoisomer. The greatest discrimination of all the tissues examined was found to occur in the brain. After five months, the level of the deuterated natural diastereoisomer was more than five times that of the deuterated 2 S-epimer. These results have potential implications for human nutrition. © 1985 American Oil Chemists' Society.