Investigation of Antioxidant Synergisms and Antagonisms among Phenolic Acids in the Model Matrices Using FRAP and ORAC Methods

The total antioxidant potential of a sample cannot be predicted from the antioxidant activity of its compounds; thus, scientists usually explain the overall activity through their combined effects (synergistic, antagonistic, or additive). Phenolic compounds are one of the most powerful and widely investigated antioxidants, but there is a lack of information about their molecular interactions. This study aimed to investigate the individual and combined antioxidant activity of equimolar mixtures (binary, ternary, quaternary, and quinary) of 10 phenolic acids (protocatechuic, gentisic, gallic, vanillic, syringic, p-coumaric, caffeic, ferulic, sinapic, and rosmarinic acid) at different concentrations using ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC) assays. Gallic acid showed the highest antioxidant activity, determined using the FRAP assay (494–5033 µM Fe(2+)) and rosmarinic acid with the ORAC assay (50–92 µM Trolox Equivalents (TE)), while the lowest antioxidant potential was observed for p-coumaric acid (FRAP 24–113 µM Fe(2+) and ORAC 20–33 µM TE). The synergistic effect (by FRAP) in the equimolar mixtures of hydroxybenzoic acids was confirmed for a large number of tested mixtures, especially at low concentrations. All mixtures containing gentisic acid showed a synergistic effect (28–89% difference). Using the ORAC method, only two mixtures of hydroxybenzoic acids showed an antagonistic effect, namely a mixture of gentisic + syringic acids (−24% difference) and gallic + vanillic acids (−30% difference), while all other mixtures showed a synergistic effect in a range of 26–236% difference. Among mixtures of hydroxycinnamic acids, the highest synergistic effect was observed for the mixtures of p-coumaric + ferulic acids and caffeic + sinapic acids with differences of 311% and 211%, respectively. The overall antioxidant activity of phenolic acids could be explained by the number or position of hydroxyl and/or methoxy functional groups as well as the compound concentration, but the influence of other parameters such as dissociation, intramolecular hydrogen bonds, and electron donating or withdrawing effect should not be neglected.