*2.4. Assessment of Total Phenol Content*

The determination of the total phenolic content (TPC) was carried out based on the method developed by Singleton and Rossi [22] adapted to bee pollen. 1 mL of the bee pollen extract solution (0.01 g/mL) was dissolved with 1 mL of Folin-Ciocalteu reagent and 10 mL of distilled water. After gently stirring and standing for 2 min, 4 mL of 7% Na2CO3 solution was added and made up to 25 mL with distilled water. The solutions were measured using a UV–Vis spectrophotometer (Jenway 6305, Fisher Scientific, Loughborough, UK) at an absorbance of 765 nm after being kept in the dark for 1 h. Gallic acid solutions were used to obtain the calibration curve. The results of TPC as gallic acid in mg/100 g were expressed.
