Please check the following links and PDF attachments.
https://www.researchgate.net/publication/12621651_Methods_to_measure_the_antioxidant_activity_in_plant_material_A_comparative_discussion
https://www.researchgate.net/post/What_is_a_simple_method_to_determine_the_antioxidant_activity_of_plant_material_extracts
https://www.ncbi.nlm.nih.gov/pubmed/10694046
https://www.researchgate.net/post/Which_is_the_best_method_to_determine_the_total_antioxidant_capacity_of_purified_plant_extracts
https://www.researchgate.net/post/How_to_calculate_antioxidant_activity_of_medicinal_plants
https://www.researchgate.net/post/How_can_I_measure_the_antioxidant_activity_DPPH_for_crude_plant_extract
Good Luck
Article Methods to measure the antioxidant activity in plant materia...
Please check the following links and PDF attachments.
https://www.researchgate.net/publication/12621651_Methods_to_measure_the_antioxidant_activity_in_plant_material_A_comparative_discussion
https://www.researchgate.net/post/What_is_a_simple_method_to_determine_the_antioxidant_activity_of_plant_material_extracts
https://www.ncbi.nlm.nih.gov/pubmed/10694046
https://www.researchgate.net/post/Which_is_the_best_method_to_determine_the_total_antioxidant_capacity_of_purified_plant_extracts
https://www.researchgate.net/post/How_to_calculate_antioxidant_activity_of_medicinal_plants
https://www.researchgate.net/post/How_can_I_measure_the_antioxidant_activity_DPPH_for_crude_plant_extract
Good Luck
Article Methods to measure the antioxidant activity in plant materia...
Dear
Several methods are known. DPPH, ABTS are most accurate than reducing power, hydrogen peroxide. However you can use them all to confirm your results. Metal chelating methodologies also could support the results.
Good luck
Methods to measure the antioxidant activity in plant material. A comparative discussion.
Arnao MB1, Cano A, Acosta M.
Abstract
Methods to determine total antioxidant activity (TAA) are generally based on the inhibition of certain reactions by the presence of antioxidants. The most widely used methods are those that involve the generation of radical compounds, and it is the presence of antioxidants that determines the disappearance of these radicals. Strategies for measuring TAA are discussed, particularly the different methodological and instrumental approaches used. In addition, our own methods are presented in order to facilitate and speed up the manipulation of biological material. The values of TAA by different compounds are presented and compared.
https://www.ncbi.nlm.nih.gov/pubmed/10694046
Dear Anil Kumar Thakur, If you study with plants, firstofall you should know that the anti-oxidant properties of plants vary under different circumstances. That is, even if you apply the same protocol in articles, you can not achive the same results given another articles.
As per the research experience that I have regarding determining the anti-oxidative potential of medicinal plants, performing the following experiments might be very useful to you:
1) DPPH scavenging assay
2) Measuring the level of Catalase and SOD
3) Measure Lipid Per oxidation (Also called as TBARS assay)
4) Chelating Assay.
As I have said I'm not as experienced as others who have replied. So kindly take it as a suggestion and cross check. However, I think the above listed experiments if what you need.
Dear Please check the following methods:
DPPH free radical scavenging assay
The DPPH free radical scavenging potential was determined according to the procedure used by Uddin et al.16 0.2 mL of different extracts (1 mg/mL) was added to 0.004% methanolic solution of DPPH (3 mL). The different samples were incubated for 30 min in the dark at room temperature and the absorbance was noted at 517 nm on UV-Visible spectrophotometer 1800 (Shimadzu). The percentage inhibition (I%) of DPPH free radical was evaluated using the following equation:
I% [DPPH free radical] = [(AC – AS) / AC] × 100
AC - the absorbance of the control and AS - the absorbance of the samples/standard solutions. The ascorbic acid and quercetin dihydtrate were used as standards. The percentage inhibition of each sample was evaluated in triplicate.
FRAP assay
The ferric ion reducing antioxidant power of different extracts and standard was measured according to the method used by Benzie and Strain.18 0.2 mL of different extracts (0.5 mg/mL) and standards (60-300 mg/L) were mixed with 2.8 mL of FRAP solution [300 mM acetate buffer (25 mL), 10 mM 2,4,6-tripyridyl-s-triazine in 40 mM HCl (2.5 mL) and 20 mM FeCl3·6H2O (2.5 mL)]. Ferrous sulphate was used as standard. The absorbance was measured at 593 nm on UV-Visible spectrophotometer after the incubation of solutions for 30 min in the dark. The results were expressed as mg of ferrous II equivalent (Fe (II) E) /1g of DPE. Each experiment was carried out in triplicate.
TAC assay
The total antioxidant capacity was evaluated according to phosphomolybdenum reducing method used by Prieto et al.19 In this assay 0.3 mL of different extract (1 mg/mL) or standard (60-300 mg/L) was taken in a test tube. After this, the reagent solution (3 mL; 0.6 M sulfuric acid, 28 mM sodium phosphate and 4 mM ammonium molybdate) was added to each test tube. All the test tubes were incubated at 95 0C for 90 min. Upon cooling to room temperature, the absorbance of the solutions was noted at 695 nm on UV-Visible spectrophotometer. The results were represented in mg of ascorbic acid equivalent (AAE) /1 g of DPE. Again, each experiment was carried out in triplicate.
Hello Anil,
Below attached manuscript may helpful to your research, it having detail procedure for antioxidant analysis by different methods.
Wish you all best wishes for your research.
Article Phytochemical, antioxidant and antibacterial activity of bla...
Methods to determine total antioxidant activity (TAA) are generally based on the inhibition of certain reactions by the presence of antioxidants. The most widely used methods are those that involve the generation of radical compounds, and it is the presence of antioxidants that determines the disappearance of these radicals. Strategies for measuring TAA are discussed, particularly the different methodological and instrumental approaches used. In addition, our own methods are presented in order to facilitate and speed up the manipulation of biological material. The values of TAA by different compounds are presented and compared.
Article Methods to measure the antioxidant activity in plant materia...
You should also know which one is most suitable for you. Biologically, antioxidants play their health-beneficial roles via transferring a hydrogen (H) atom or an electron (e–) to reactive species, thereby deactivating them. Antioxidant activity assays imitate this action; that is, antioxidants are measured by their H atom transfer (HAT) or e– transfer (ET) to probe molecules. Antioxidant activity/capacity can be monitored by a wide variety of assays with different mechanisms, including HAT, ET, and mixed-mode (ET/HAT) assays, generally without distinct boundaries between them.
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