Abstract
Size-exclusion chromatography (SEC) and activity-guided fractionation of honeys allowed the isolation of high molecular weight brown compounds, ranging in size from 66 to 235kDa that exhibited peroxyl radical-scavenging activity. Their concentrations, antioxidant activity and degree of browning increased after heat-treatment of honeys, suggesting that they represent melanoidins. Chemical analysis of melanoidins demonstrated the presence of proteins, polyphenols and oligosaccharides. Heat-treatment caused an increased incorporation of phenolics into high molecular weight melanoidins and drastically decreased the protein content in these fractions with a concomitant appearance of high molecular weight protein–polyphenol complexes of reduced solubility. LC–ESI–MS demonstrated the presence of oligosaccharide moieties, supporting the postulated origin of melanoidins. The changes in the phenolic content of melanoidins from heated honeys were strongly correlated with their oxygen radical absorbance capacity (ORAC) values (R=0.75, p<0.0001), indicating that polyphenols contribute to the antioxidant activity of melanoidins. In summary, honey melanoidins are multi-component polymers consisting of protein–polyphenol–oligosaccharide complexes. A direct interaction between polyphenols and melanoidins resulted in a loss or gain of function for melanoidin antioxidant activity.
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