Determination of phenolic compounds and their antioxidant activity in fruits and cereals

Three methods, FCM (with Folin–Ciocalteu reagent), PBM (Price and Butler) and AAPM (with 4-aminoantipyrine) for assessment of phenolic compounds and three commonly used methods, TEAC (Trolox equivalent antioxidant capacity), DPPH (with diphenyl-picrylhydrazyl radical), and FRAP (ferric reducing antioxidant power) for evaluation of antioxidant capacity, were modified to a semimicroscale (total volume 1 ml) with minimum consumption (to 100 μl) of a sample and thereby applicable for fast screening. Appropriate standards and extracts of 17 kinds of fruit and six kinds of cereal were assessed for total content of phenolic compounds and total antioxidant capacity by each of these methods. The results of analyses of commonly used standards (gallic, caffeic and ferulic acids, (+)-catechin, Trolox, fenol and FeSO4) for these methods and identical plant extract showed different reactivity of principal reagent of the methods with individual standards and therefore with phenolic substances of extracts as well. However, the trends of the measured values of extracts could be compared, though their absolute values differ proportionally. At assessments of phenolic compounds it is important to determine content of ascorbic acid at roughly the same time and correct the obtained values according to its contribution to the increase in absorbance calculated on the basis of absorbance equations, especially for samples with a higher content. The same is true for reducing saccharides; they can significantly “elevate” values of contents of phenolic compounds and antioxidant activities (by even more than 50%), especially in samples of sweeter fruits. The saccharides should therefore be removed or a correction applied reflecting their concentration.