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Potency of Antioxidant Properties of Major Secondary Metabolite ā€ˇfrom Stevia rebaudiana in a Comparison to Synthetic Antioxidants

Amal Ali Awadalla Mohamed, Mostafa E Rateb, marcel Jaspars.

Abstract
Bioactivity-guided fractionation was performed to evaluate the ā€ˇantioxidant activity of Stevia rebaudiana Bertoni. The methanolic crude ā€ˇā€ˇ(MC) extract of leaves was fractionated using a modified Kupchan ā€ˇpartitioning method. The antioxidant activity was measured for the MC ā€ˇand fractions. Free radical scavenging using 2,2-Diphenyl-1-picryl-ā€ˇhydrazyl (DPPH) and total antioxidant activity using ā€ˇphosphomolybdenum complex methods were selected for ā€ˇdetermination of the antioxidant activity using ascorbic acid (AA) as a ā€ˇreference. Preliminary antioxidant activity measurements revealed that ā€ˇthe dichloromethane fraction was of interest. Consequently, it was ā€ˇsubjected to chromatographic separation and purification techniques. ā€ˇOne major pure compound was isolated and its structure elucidation ā€ˇusing NMR and HRMS confirmed it to be 3',5,7-trihydroxy-3,4',6-ā€ˇtrimethoxyflavone (centaureidin). The IC50 values of MC, butanol, ā€ˇmethanol and dichloromethane fractions and centaureidin (CT) were ā€ˇā€ˇ120.6±0.58, 66.5±0.49, 79.5±0.44, 37.1±0.23 and 18.4±0.44 µg/mL, ā€ˇrespectively. The results were compared to those of synthetic ā€ˇantioxidants, butylated hydroxyanisole (BHA) and butylated ā€ˇhydroxytoluene (BHT) with IC50 values of 13.7±0.09 and 33.1±1.05 ā€ˇā€ˇµg/mL, respectively; while that of AA was 6.5±0.25 µg/mL. The highest ā€ˇtotal antioxidant activity evaluated by the phosphomolybdenum ā€ˇcomplex method was exhibited by the purified compound (CT) with a ā€ˇvalue of 5028.6±645.87 mg AA equivalent/g sample. The study ā€ˇreveals that components of this plant could be a potential alternative to ā€ˇsynthetic antioxidants by virtue of their antioxidant properties and ā€ˇnatural origin.ā€ˇ

Key words: antioxidant activity; Asteraceae; Stevia rebaudiana; synthetic ā€ˇantioxidant; natural antioxidant; Centaureidin


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