2203 uv vis spectrophotometer

Scavenging activity on DPPH free radicals by the extracts was assessed according to the method reported with slight modifications [20 , 21 ]. Briefly, a 2.0 ml solution of the extract, at different concentrations diluted two-fold (2–125 μg/ml) in methanol was mixed with 1.0 ml of 0.3 mM DPPH in methanol. The mixture was shaken vigorously and allowed to stand at room temperature in the dark for 25 min. Blank solutions were prepared with each test sample solution (2.0 ml) and 1.0 ml of methanol while the negative control was 1.0 ml of 0.3 mM DPPH solution plus 2.0 ml of methanol. Thereafter, the absorbance of the assay mixture was measured at 518 nm against each blank with Systronics 2203 UV–vis spectrophotometer. Lower absorbance of the reaction mixture indicated higher radical-scavenging activity. BHA was used as positive control. DPPH radical-scavenging activity was calculated using the equation;
$\mathrm{D}\mathrm{P}\mathrm{P}\mathrm{H}\mathrm{\%}=(\mathrm{A}\mathrm{b}\mathrm{l}\mathrm{a}\mathrm{n}\mathrm{k}-\mathrm{A}\mathrm{s}\mathrm{a}\mathrm{m}\mathrm{p}\mathrm{l}\mathrm{e})/\mathrm{A}\mathrm{b}\mathrm{l}\mathrm{a}\mathrm{n}\mathrm{k})\times 100$
where A blank is the absorbance of the control reaction (containing all reagents except the sample) and A sample is the absorbance of test sample (with the DPPH solution).

The method suggested by Wilcox et al. [21 (link)] was employed for seed extraction with HCl (0.4 mM) for phytic acid evaluation. To obtain a clear supernatant, Chen’s reagent (3 M sulfuric acid, 2.5% ammonium molybdate, 10% ascorbic acid and deionized water in the ratio of 1:1:1:2) was utilized. The reading was recorded at a 450 nm wavelength by employing a Systronics 2203 UV-Vis spectrophotometer (Systronics, Ahmedabad, India).

Scavenging activity on DPPH free radicals by the extracts was assessed according to the method reported by Awah et al. [14 (link)] with slight modifications of Gyamfi et al. [15 (link)]. Briefly, a 2.0 mL solution of the extract, at different concentrations diluted twofold (2–125 μg/mL) in methanol, was mixed with 1.0 mL of 0.3 mM DPPH in methanol. The mixture was shaken vigorously and allowed to stand at room temperature in the dark for 25 min. Blank solutions were prepared with each test sample solution (2.0 mL) and 1.0 mL of methanol while the negative control was 1.0 mL of 0.3 mM DPPH solution plus 2.0 mL of methanol. L-ascorbic acid was used as the positive control. Thereafter, the absorbance of the assay mixture was measured at 518 nm against each blank with Systronics 2203 UV-Vis spectrophotometer. Lower absorbance of the reaction mixture indicated higher radical scavenging activity.
DPPH radical scavenging activity was calculated using the equation $\begin{array}{c}\mathrm{D}\mathrm{P}\mathrm{P}\mathrm{H}\%=\left(\frac{\left(A_{\mathrm{b}\mathrm{l}\mathrm{a}\mathrm{n}\mathrm{k}}-A_{\mathrm{s}\mathrm{a}\mathrm{m}\mathrm{p}\mathrm{l}\mathrm{e}}\right)}{A_{\mathrm{b}\mathrm{l}\mathrm{a}\mathrm{n}\mathrm{k}}}\right)\times \mathrm{100}.\end{array}$