Determination of Antioxidant Activity in Foods and Beverages by Reaction with 2,2′-Diphenyl-1-Picrylhydrazyl (DPPH): Collaborative Study First Action 2012.04

Abstract A colorimetric method for the determination of total antioxidant activity in a variety of foods and beverages was validated in both a single-laboratory validation and a collaborative laboratory validation study. The procedure involved extraction of the antioxidants directly into a methanol–water solution containing a known amount of 2,2′-diphenyl-1-picrylhydrazyl (DPPH), thus promoting the rapid reaction of extracted materials with DPPH. The reaction was monitored by spectrophotometric measurement of the absorbance loss at 517 nm. Antioxidant activity was quantified relative to a dilution series of vitamin E analog standards (Trolox®), which were analyzed in parallel simultaneously with the food and beverage samples. The antioxidant activities of the samples ranged from 131 to 131 000 μmole Trolox equivalents/100 g. Statistical analysis of the results showed that nine of the 11 matrixes gave acceptable HorRat values, indicating that the method performed well in these cases. The acceptable matrixes include pomegranate juice, blueberry juice, carrot juice, green tea, wine, rosemary spice, ready-to-eat cereal, and yogurt. Two samples failed the HorRat test: the first was an almond milk that had an antioxidant level below the practical LOQ for the method; the second was a sample of canola oil with added omega-3 fatty acid that was immiscible in the reaction medium.