I need to estimate antioxidant in lichens methanolic and acetonic extract samples , by using FRAP method, I'm just searching an easy protocol or any paper help me
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Article Purification and characterization of fucose-containing sulph...
Ferric reducing antioxidant power assay The ferric reducing activity of sulphated polysaccharides was carried out based on the method of Benzie and Strain (1996). The solutions for this assay consisted of 300 mM L−1 acetate buffer (pH), 10 mM L−1 (TPTZ) 2,4,6-tripyridyl-s-triazine in 40 mM L−1 of HCl and 20 mM L−1 FeCl3.6H2O. The freshly prepared ferric reducing antioxidant power (FRAP) reagent was incubated at 37 °C for 5 min. 0.05 mL of sulphated polysaccharides at 1 mg mL−1 was added to 0.95 mL of FRAP reagent. The reagent mixture was incubated at 37 °C for 30 min. The absorbance was read at A593nm. The results were expressed as μM of Fe (II) reducing activity per mg of dry weight of sulphated polysaccharides. A calibration curve was plotted using various concentrations of FeSO4·7H2O (100– 400 μM)
The FRAP (ferric reducing antioxidant power) method: The FRAP (ferric reducing antioxidant power) method relies on the reduction by the antioxidants, of the complex ferric ion-TPTZ (2,4,6-tri(2-pyridyl)- 1,3,5-triazine). The binding of Fe2+ to the ligand creates a very intense navy blue color. The absorbance can be measured to test the amount of iron reduced and can be correlated with the amount of antioxidants . Trolox or ascorbic acid
You know what is the absorbance of the FRAP solution? or what is the blank to calibrate and take the control absorbance?
The reduced FRAP reagent will give blue colour, so can read at 593 nm. You can keep FRAP reagent without any sample or standard as blank
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