Simple colorimetric method to determine the in vitro antioxidant activity of different monoterpenes

The development of simple, fast and reproducible techniques that provide information about the antioxidant activity (AA) of different compounds is essential to screen and discover new molecules with potential applications in the therapeutic, cosmetic, toxicological and food fields. Here, a novel and simple colorimetric method ("BCB assay") is proposed for measuring the AA of chemical compounds by protection of the reporter dye Brilliant Cresyl Blue (BCB) from loss of color due to oxidation by hypochlorite (a physiological oxidant). The decay in BCB blue color (λ_max = 634 nm) in the presence of hypochlorite occurred in only 5 min and was used to track the AA of different molecules. Particularly, the AA of monoterpenes was demonstrated and used to quantify them at milimolar concentrations. Natural antioxidants like vitamins C and E, resveratrol, dithiothreitol, N-actyl-l-cysteine and glutathione were used as controls to validate the assay. Linalool, geraniol and 1,8-cineole were tested and showed in vitro AA in a concentration-dependent manner. The monoterpene concentrations providing 50% protection against oxidation (AA₅₀) were 2.3, 36.2 and 135.0 mM for linalool, geraniol and 1,8-cineole respectively, suggesting interesting AA. The method provides a useful, fast, simple and low-cost tool to determine the in vitro AA of different molecules.