2.6.4. Hydrogen peroxide scavenging assay

The H₂O₂-scavenging activities were analyzed using a chemiluminescence method [24]. A carbonate buffer solution (pH 9.5) with a concentration of 50 mmol/L and a luminol solution of 1 mmol/L were mixed at a ratio of 17:1 away from light (working solution). For the measurement, 10 μL of different concentrations of the spent substrate extract of C. militaris, 10 μL of a 6% H₂O₂ solution, and 150 μL of the luminol-carbonate system luminescence working solution were added to the a 96-well plate, and the chemiluminescence was measured. The reaction time was set to 30 s, and the experimental data were measured every 0.3 s to obtain the highest value, A1, with deionized water as a blank control. The peak control value was recorded as A0.

Three replicate wells were used for in each group and the average value was obtained. The formula for hydrogen peroxide anion clearance was: