A thorough study was done to test the reactivity of the Folin-Ciocalteu (F-C) reagent toward various compound classes. Over 80 compounds were tested. Compound classes included phenols, thiols, vitamins, amino acids, proteins, nucleotide bases, unsaturated fatty acids, carbohydrates, organic acids, inorganic ions, metal complexes, aldehydes, and ketones. All phenols, proteins, and thiols tested were reactive toward the reagent. Many vitamin derivatives were also reactive, as were the inorganic ions Fe(+2), Mn(2+), I(-), and SO(3)(2-). Other compounds showing reactivity included the nucleotide base guanine and the trioses glyceraldehyde and dihydroxyacetone. Copper complexation enhanced the reactivity of salicylate derivatives toward the reagent, whereas zinc complexation did not. Several amino acids and sugars that were reported to be reactive toward the F-C reagent in earlier studies were found not to be reactive in this study, at least in the concentrations used. Reaction kinetics of each compound with the F-C reagent were also measured. Most compounds tested showed a biphasic kinetic pattern with half-lives under 1 min. Trolox and ascorbic acid displayed a rapid monophasic pattern in which the reaction reached end point within 1 min. In summary, this study has shown that the F-C reagent is significantly reactive toward other compounds besides phenols. As other investigators have suggested, the F-C assay should be seen as a measure of total antioxidant capacity rather than phenolic content. Because phenolics are the most abundant antioxidants in most plants, it gives a rough approximation of total phenolic content in most cases.