2.7.1. Amplex Red Hydrogen Peroxide Assay

Hydrogen peroxide levels present in CFCS were measured using the Amplex Red Hydrogen Peroxide/Peroxidase kit according to the manufacturer's recommendations (Thermo Fisher Scientific, Waltham, USA). After preparing the kit stock solutions, aliquots (50 μL) of the standard curve samples, controls, and experimental samples were added to individual wells on a microplate. After that, the Amplex® Red reagent/HRP working solution (50 μL) was added to the wells previously plotted. After incubation (30 min, room temperature, protected from light), absorbance was measured in a microplate reader (550 nm) to construct the standard curve and measure the H₂O₂ concentration (μM) of the CFCS.