were evaluated using determination of DPPH free radical scavenging activity, determination of reducing power, determination of ferrous ion chelating ability, click here and total phenolic content determination methods. The stable DPPH radical scavenging activity
was measured using the modified method described by Chang et al11 Stock solution (1 mg/ml) of the ethanol extract of the leaves of A. conyzoides and M. cordifolia were prepared in ethanol from which serial dilutions were carried out to obtain the concentrations of 5, 10, 20, 40, 60, 80, 100 μg/ml. In this assay, 2 ml of 0.1 mm ethanolic DPPH solution was added to 2 ml of extract solution at different concentrations and the contents were stirred vigorously for 15 s. Then the solutions were allowed to stand at dark place at room temperature for 30 min for reaction to occur. After 30 min, absorbance was measured against a blank at 517 nm with a double beam UV spectrophotometer (UV-1800, Shimadzu, Japan). The percentage of DPPH radical scavenging activity of each plant extract
was calculated as: DPPHradical−scavengingactivity(I%)=[(A0−A)/A0]×100where A0 is the absorbance of the control solution (containing all reagents except plant extracts); A is the absorbance of the DPPH solution containing plant extract. The concentration of sample required to scavenge 50% DPPH free radical (IC50) was calculated from the plot of inhibition (%) against the concentration Quizartinib ic50 of the extract. Ascorbic acid and BHA were used as positive control standard. This assay was determined according to the method reported by Oyaizu12 with slight modifications. Briefly, 1 ml of extract solution of different concentrations (5, 10, 20, 40, 60, 80, 100 μg/ml) was mixed with 2.5 ml of phosphate buffer (0.2 M, pH 6.6) and 2.5 ml of potassium ferricyanide [K3Fe(CN)6] (1% w/v). The mixture was incubated at 50 °C for 20 min. The reaction was terminated by adding 2.5 ml of Trichloroacetic acid (10%, w/v), then the mixture was centrifuged at 3000 rpm for 10 min. The supernatant solution (2.5 ml) was mixed with distilled water (2.5 ml)
and ferric chloride (0.5 ml, 0.1% w/v) solution. Then the absorbance was measured at 700 nm against a blank using UV spectrophotometer. Increased absorbance value of the reaction mixture Vasopressin Receptor indicates increased reducing power. Three replicates were made for each test sample and average data was noted. Here, ascorbic acid and BHA were used as positive control standard, too. The ferrous ions chelating activity of ethanol extract of the leaves of A. conyzoides and M. cordifolia, and standards was investigated according to the method of Dinis et al 13 Briefly, ethanol extracts (5 ml) was added to 0.1 ml solution of 2 mM FeCl2 and ethanol. Then, the reaction was initiated by the addition of 0.2 ml of 5 mM ferrozine and mixture was shaken vigorously and left standing at room temperature for 10 min.
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