Antioxidant HPTLC-DPPH Fingerprinting of Honeys and Tracking of Antioxidant Constituents upon Thermal Exposure

The use of High-Performance Thin-Layer Chromatography (HPTLC) coupled with the use of DPPH* (2,2-diphenyl-1-picrylhydrazyl) as a derivatisation reagent is a novel approach to the analysis of antioxidant activity of honeys. The method facilitates the visualisation of individual constituents that contribute to the overall antioxidant activity of the honey, even if they are not yet chemically identified, and allows for the quantification of their antioxidant activity as gallic acid equivalents. The method supports a more in-depth study of the antioxidant activity of honey as it allows for a comparative analysis of the antioxidant fingerprints of honeys of different floral origin and is able to capture differences in their individual bioactive constituents. Further, it supports the tracking of changes in antioxidant activity of individual honey constituents over time upon exposure to different temperature conditions, which demonstrates the potential value of the method for in-process quality control.