Oxidation of lipid, nucleic acids or protein has been suggested to be involved in the etiology of several chronic diseases including cancer, cardiovascular disease, cataract, age-related macular degeneration and aging in general. A large body of research has investigated the potential role of antioxidant nutrients in the prevention of these and other chronic diseases. This review concentrates on the following antioxidant nutrients: beta-carotene and other carotenoids, vitamin E, vitamin C and selenium. The first part of the review emphasizes the utility of biological markers of exposure for these nutrients and the relationship to dietary intake data. The second part considers functional assays of oxidative stress status in humans including the strengths and limitations of various assays available for use in epidemiologic research. A wide variety of functional assays both in vivo and ex vivo, are covered, including various measures of lipid oxidation (thiobarbituric acid reactive substances, exhaled pentane/ethane, low-density lipoprotein resistance to oxidation, isoprostanes), DNA oxidation (oxidized DNA bases such as 8-OHdG, autoantibodies to oxidized DNA, modified Comet assay) and protein oxidation (protein carbonyls). Studies that have examined the effects of antioxidant nutrients on these functional markers are included for illustrative purposes. The review concludes with a discussion of methodologic issues and challenges for studies involving biomarkers of exposure to antioxidant nutrients and of oxidative stress status.