2.4. Total Phenolic Content Determination

Total phenolic content (TPC) determination was carried out following the protocol described by Ainsworth and Gillespie [13]: 100 µL of phenolic extracts were mixed with 200 µL of 10% (v/v) Folin-Ciocalteu’s reagent and incubated for 2 min at room temperature in the dark. Next, 800 µL of 0.7 M sodium carbonate was added, and the samples were vortexed and incubated for 2 h at 25 ± 1 °C in the dark. The absorbance was measured at λ = 765 nm against a blank and a calibration curve was performed using gallic acid as standard. Results were expressed as gallic acid equivalents (GAE) in mg/g DW. All determinations were carried out in triplicate.