A novel fluorescent ORAC-SIA method to determine antioxidant capacity in several food samples using fluorescein as the probe was developed. The optimization of the method was through a multivariable design, decreasing the analysis time to 5 min and the AAPH concentration to 67% compared with 90 min in the standard 96-well microplate method. The aspiration order was AAPH-sample/standard-fluorescein injected into a stream of a water-based carrier. The calculation of the antioxidant capacity was done from the fluoresceine peak heigh, so neither delay time nor area measurement was necessary. The proposed method showed excellent precision (RDS < 3%) with a LOD of 3.13 µmol L-1 and recoveries from 90% to 107%. The results from the ORAC-SIA method did not show a significant difference from the microplate method.
Keywords: AAPH; Antioxidant capacity; Fluorescein; Food samples; ORAC; SIA.
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