Optimization and validation of an alternative method to evaluate total reactive antioxidant potential.

The total reactive antioxidant potential (TRAP) is one of the methods most employed to estimate the antioxidant capacity of samples in vitro. This method is based on the quenching of luminol-enhanced chemiluminescence derived from the thermolysis of 2,2'-azo-bis(2-amidinopropane)dihydrochloride (AAPH) as the free radical source. However, this method can present limitations when the sample does not present a lag phase. In addition, there are no studies regarding TRAP assay validation. In this context, the aim of this work was to optimize and validate this method and to propose another evaluation method using the area under the curve (AUC). The main condition established was the need for the stabilization of the system, at 7000s, before the addition of the antioxidant to be tested. Both evaluation methods were validated using Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) as a calibrator in the range of 50 to 250nM, and all parameters showed satisfactory results: specificity, linearity (r>0.99), precision (intra and interassay relative standard deviations <5%), robustness, and the limits of detection and quantitation (low and similar for both methods). The main advantage of the use of AUC is to evaluate the antioxidant potential of samples that do not present lag phase.

Antioxidant potential of peels and fleshes of peaches from different cultivars.

Increasing recent interest in nutraceuticals and functional foods has led researchers to investigate the antioxidant potential of several fruits. This article evaluates the antioxidant potential and reactivity based on luminol-enhanced chemiluminescence capacity of peach extracts (peels and fleshes) and the contribution of a major compound present in these extracts to antioxidant potential and reactivity. The results obtained showed that the extracts of peels and fleshes of Maciel, Leonense, and Eldorado peach cultivars present free radical scavenging activity in all concentrations tested, with a concentration-dependent action. The immediate inhibition of chemiluminescence and the duration of this inhibition were significantly higher with the extracts than with the major compound (chlorogenic acid) alone, and it can be due to a synergistic or additive effect of other antioxidants present in the extracts. The 50% inhibitory concentration (IC(50)) values for peach extract and chlorogenic acid were 1.19 microg/mL and 8.43 microg/mL, respectively, when total radical-trapping antioxidant potential was evaluated, whereas IC(50) values of 0.41 microg/mL and 1.89 microg/mL was found when total antioxidant reactivity was evaluated in peach extract and chlorogenic acid, respectively. Chlorogenic acid presented a good contribution to antioxidant reactivity and potential, but the fruit extracts provide better antioxidant action. Peach could be of great interest as an important antioxidant source including chlorogenic acid, and it may provide health-promoting advantages to consumers by intake of this fruit or by utilization of its peels as antioxidant sources in industry.