*3.4. Antioxidant Analyses*

All the antioxidant analyses of the extracts were performed in triplicate (*n* = 3).

#### 3.4.1. 1,1-Diphenyl-2-Picryl-Hydrazil (DPPH) Radical Scavenging Activity

DPPH assay was performed following the protocol as described by Sridhar and Charles [53]. A total of 700 μL of a methanolic DPPH solution (100 μM) was added to 700 μL of sample/standard, incubating the mixtures at room temperature in the dark for 20 min. The absorbance of the reactions was measured against blank (methanol without DPPH solution) at a wavelength of 515 nm in a spectrophotometer. Trolox (97% purity) was used as standard at concentrations ranging from 10–150 mM/L. DPPH results were expressed as mM TE/g.

#### 3.4.2. Ferric Reducing Antioxidant Power (FRAP) Assay

FRAP assay was performed following the method described by Benzie and Strain [54]. Fresh FRAP working solutions were prepared by mixing 300 mM acetate buffer, pH 3.6; 10 mM 2,4,6-Tri(2-pyridyl)-s-triazine in 40 mM hydrochloric acid and 20 mM Iron(III) chloride hexahydrate in the ratio of 10:1:1, *v/v*/*<sup>v</sup>*. A total of 2.5 mL of pre-heated FRAP working solution (37 ◦C) was added to 83 μL of sample/standard and the mixtures were incubated 10 min in dark conditions at room temperature. The absorbance of the reactions was measured at 593 nm in a spectrophotometer against a reagen<sup>t</sup> blank (FRAP working solution). Trolox was used as standard at concentrations of 10–150 mM/L. FRAP results were expressed as mM TE/g.
