The thiobarbituric acid reactive substances (TBARS) assay is widely used to measure lipid oxidation and antioxidant activity in food and physiological systems. However, there has been no review (to our knowledge) that focuses exclusively on this test. This review presents an overview of the current use of the TBARS test in food and physiological systems, before looking at the various ways in which the assay is used in studies on antioxidant activity. As an antioxidant assay, the TBARS test may lack acceptable reproducibility, and long reaction times may preclude its adoption as a rapid screening method. Despite these potential limitations, there are features of the TBARS test that make it useful as a complement to popular screening tests such as Trolox equivalent antioxidant capacity. This review concludes with proposals for development of the TBARS test so that it can be used as a rapid and robust antioxidant assay.