*2.6. Radical-Scavenging Activity*

The method used to evaluate the antioxidant activity of honey is based on the discoloration of a 2,2-diphenyl-1-picrilhidrazil (DPPH) solution. The methodology measures the radical-scavenging activity of a honey solution against DPPH by spectrophotometry [26]. A solution (0.1 g/mL) of each honey sample in methanol was prepared. Then, 0.3 mL of the honey solution was mixed with 2.7 mL of the DPPH solution (6 × 10−<sup>5</sup> M); a blank was also prepared. The honey sample solution and the blank were maintained in the dark at room temperature for 30 min. Then, absorbance at 517 nm was measured with a UV-vis spectrophotometer (Jenway 6305, Staffordshire, UK). Radical Scavenging Activity (RSA) percentage was calculated considering DPPH discoloration for each sample, tested as follows: RSA = [(AbsB − AbsS)/AbsB] × 100, where AbsB is the absorbance of the blank, and AbsS is the absorbance of the honey sample solution.
