A new bioassay using phage is described for the assay of antioxidative activity. The assay is based on: (1)
in vitro in aqueous solution oxygen radicals being produced during oxidation of ascorbic acid and related substances, (2) phage being inactivated by the oxygen radicals, and (3) substances having antioxidative activity inhibiting the inactivation of phage. Samples were ground with a pestle, then diluted with water. The assay method is as follows. Phage T2 (4×10
7PFU/mL) was incubated with iron (II)-erythorbate complex (1×10
-6M) and each sample in 0.02M Tris-HC1 buffer (pH 7.4) for 20min at 37°C. The phage inactivation of a control (sample-free) is 90%. Unit activity is defined as the reciprocal of the highest dilution that shows 50% inhibition of the phage inactivation.
The effectiveness of the assay method was confirmed using known antioxidants. Then the activity was effectively assayed for many vegetables and fruits.
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