I want to know the proper protocol and mechanism of Fe++ metal chelating assay for the determination of antioxidant scavenging activity of the plant extracts and what solvent is used for the chemical used in this assay and how to calculate its ic50 and its final result. Can anybody help me for this?
HI Verma,
Briefly, the extract (from 25 - 75 μg/mL) in 0.4 ml was added to a solution
of 2 mM FeCl2 (0.2 ml). The reaction was initiated by the addition of
5 mM ferrozine (0.4 ml) and total volume was adjusted to 4 ml of
ethanol. Then, the mixture was shaken vigorously and left at room
temperature for ten minutes. Absorbance of the solution was then
measured spectrophotometrically at 562 nm.
Dinis TCP, Madeira VMC, Almeida LM (1994). Action of phenolic
derivates (acetoaminophen, salycilate and 5-aminosalycilate) as
inhibitors of membrane lipid peroxidation and as peroxyl radical
scavengers. Arch. Biochem. Biophys. 315: 161-169.
Hi, Urvashi,
Please, look here; http://www.ajol.info/index.php/ajb/article/viewFile/59257/47555
This will help you!
The antioxidant activity is thus evaluated at various levels by using diverse in vitro assay protocols like, DPPH- Radical Scavenging method, Total Phenol and Flavanoid estimation, Ferrous ion Chelating activity, Ferric Reducing Antioxidant Power,
Thiobarbituric acid Reactive Species, Automatic Determination of the Oxidative Stability of Fat etc
HI Verma,
Briefly, the extract (from 25 - 75 μg/mL) in 0.4 ml was added to a solution
of 2 mM FeCl2 (0.2 ml). The reaction was initiated by the addition of
5 mM ferrozine (0.4 ml) and total volume was adjusted to 4 ml of
ethanol. Then, the mixture was shaken vigorously and left at room
temperature for ten minutes. Absorbance of the solution was then
measured spectrophotometrically at 562 nm.
Dinis TCP, Madeira VMC, Almeida LM (1994). Action of phenolic
derivates (acetoaminophen, salycilate and 5-aminosalycilate) as
inhibitors of membrane lipid peroxidation and as peroxyl radical
scavengers. Arch. Biochem. Biophys. 315: 161-169.
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