Oxygen radical absorbance capacity (ORAC) assay has been applied to determine the Hydrophilic – ORAC index in food. The method measures antioxidant scavenging activity against peroxyl radical induced by 2,2′-azobis(2-amidino-propane) dihydrochloride (AAPH) at 37ºC, used fluororescein as the fluorescent probe. The antioxidant capacity is measured by assemssing the fluorescence decay curve (AUC) of the sample as compared to that of the blank in which no antioxidant is present. Results expressed in ORAC units, equivalent to micromole Trolox per 100 grams sample (Trolox equivalent). The method was shown with high selectivity, a wide linear range, from 5 to 50µM Trolox with linear coefficient R2 = 0.9987. The recovery from 93.2% to 104.4% and repeatability (RSD) was less than 6.60%. The limits of detection and quantitation were 5 and 10µM Trolox, respectively. The method has been applied to determine of H-ORAC index in 65 samples including vegetables, fruits, vegetable products and health supplements with content ranging from 720 µM TE/100g to 310878 µM TE/100g.
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