2 20 azobis 2 amidinopropane

TAC, an index of the total antioxidant strength of a molecule, was measured using a water-soluble analog of vitamin E (Trolox) equivalent antioxidant capacity assay. This assay is based in a hydrogen atom transfer that measures the capability of an antioxidant to quench free radicals by hydrogen atom donation [72 (link)]. This analysis was carried out in the supernatant obtained after macerating the muscle as previously described by Romay et al. [73 (link)]. Briefly, 10 µL of supernatant were mixed with a 1:1 mixture of 150 µM 2,20-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (Sigma-Aldrich, St. Louis, MO, USA) and 10 mM 2,20-azobis(2-amidinopropane) (Sigma-Aldrich, St. Louis, MO, USA), previously incubated at 45 °C for 30 min. Radicals presents in the mixture were scavenged by the antioxidants in the sample, and as a result, the colored product concentration decreases over time. Kinetics was determined at 734 nm for 10, 30 and 50 s after adding the sample. The antioxidant capacity thus measured was compared with that of Trolox (Sigma-Aldrich, St. Louis, MO, USA) and expressed in TEAC.