The β-Carotene bleaching activity of M. nervosa EO was assessed as described previously [10 (link),51 (link)]. Briefly, 0.5 mg of β-Carotene was dissolved in 1 mL of chloroform. Then, 200 mg of tween 40 and 25 μL of linoleic acid were added to the chloroform solution. The chloroform was removed by vacuum evaporation at 40 °C. Next, 100 mL of hydrogen peroxide was introduced, and the mixture was subjected to vigorous stirring. After preparing the emulsion, 20 μL of M. nervosa EO was added to the β-Carotene/linoleic acid mixture in 96-well microtiter plates. Plates were incubated at 50 °C for 120 min, and absorbance was read at t = 0 min and t = 120 min of incubation (automated plate reader ELx 800 Biotek, Biotek, Winooski, VT, USA).
As a standard reference, the same procedure was carried out using 20 μL of BHT solution (Vigon International, East Stroudsburg, PA, USA) in a solvent. Additionally, a control solution having the same composition, but lacking β-Carotene, was prepared.
The assessment of antioxidant activity (AA) was performed based on β-Carotene bleaching and the following equation was used:
In this equation, At corresponds to the absorbance value measured for the sample tested after incubation for 120 min, Ct represents the absorbance value of the standard reference at the same time point, and C0 means the absorbance value of the standard reference measured at the initial time.
The results are presented in terms of IC50 (μg mL−1), representing the concentration required to achieve 50% inhibition of β-Carotene bleaching. To ensure accuracy and consistency, all analyses were performed in triplicate.
As a standard reference, the same procedure was carried out using 20 μL of BHT solution (Vigon International, East Stroudsburg, PA, USA) in a solvent. Additionally, a control solution having the same composition, but lacking β-Carotene, was prepared.
The assessment of antioxidant activity (AA) was performed based on β-Carotene bleaching and the following equation was used:
In this equation, At corresponds to the absorbance value measured for the sample tested after incubation for 120 min, Ct represents the absorbance value of the standard reference at the same time point, and C0 means the absorbance value of the standard reference measured at the initial time.
The results are presented in terms of IC50 (μg mL−1), representing the concentration required to achieve 50% inhibition of β-Carotene bleaching. To ensure accuracy and consistency, all analyses were performed in triplicate.
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