There are quite a number of methods to evaluate the in vitro antioxidant activities. Can you please suggest me the best and the most effective method?
Hi, Arvind, one of the way is to measure activity of several antioxidant enzymes like catalase, peroxidase, SOd etc.
But remember, that activity of one enzyme may not give you correct picture, because enzymes have different localization (catalase is peroxisomal, APX is cytoplasmic, plastidic etc).
In addition, you can also measure contents of low molecular weight antioxidants like ASC, GSH, tocopherol etc.
More information you can find in my profile.
Dear Dr. Pasternak,
Thanks for your suggestion but as per my knowledge the antioxidant enzymes you mentioned are to access the activity in vivo and not in vitro. My query was regarding in vitro methods.
Methods Enzymol. 2002;359:366-79.
Evaluation of antioxidant activity using peroxynitrite as a source of radicals.
Geletii YV, Balavoine GG.
Russian Journal of Bioorganic Chemistry
November 2002, Volume 28, Issue 6, pp 501-514
The Determination of Total Concentration and Activity of Antioxidants in Foodstuffs
Yu. V. Geletii, G. G. A. Balavoine, O. N. Efimov, V. S. Kulikova
Nitric Oxide. 1999;3(1):40-54.
Peroxynitrite scavenging by different antioxidants. Part I: convenient assay.
Balavoine GG1, Geletii YV.
Hi arvind
For evaluation of in vitro antioxidant activity of plants, you should adopt the methods of
DPPH free radical scavenging activity, nitric oxide scavenging activity, superoxide scavenging activity, hydroxyl radical scavenging activity etc.
Dear Arvind
You can follow the following methods to evaluate in vitro antioxidant activity of plants.--
1. DPPH Free radical scavenging activity
2. Determination of total phenolic content
3. Determination of total flavonoid content
4. Reducing power assay & so on
You can read the following articles:
http://www.sciencedirect.com/science/article/pii/S0974694313002387
http://pelagiaresearchlibrary.com/european-journal-of-experimental-biology/vol3-iss1/EJEB-2013-3-1-449-454.pdf
10.5897/JMPR2014.5443
http://ajp.mums.ac.ir/jufile?c2hvd1BERj0xMDU2Jl9hY3Rpb249c2hvd1BERiZhcnRpY2xlPTEwNTYmX29iPTVjYWVmYTdmZGMwM2RmNjExZDE1YjJiZmI3NTUyODBlJmZpbGVOYW1lPWZ1bGxfdGV4dC5wZGY
Dear Arvind, In-vitro antioxidant assay are done through evaluation of radical-scavenging activity by DPPH colourimetry assay; Ferric reducing antioxidant potential (FRAP) and ABTS-FRS ability.
Dear Dr. Arvind,
Based on the results of one assay of antioxidant activity, you may not the proper results, therefore you always have to perform at least more than one or two assay, to examine the radical scavenging profile of the different phytoconstitutes (because of their complex chemical nature and also mixture of different compounds in crude extracts); and most easiest assays are DPPH, TPC, TFC, FRAP and ABTS. For more information, see the files given below. All the best
Dear Arvind,
Whatever you read in answers, the best and simplest assay does not exist (see the profile of Dr Guido Haenen). Just use the assay(s) you can use in your laboratory and which you understand well. When I developed my tube assay, the simplicity (skills of labor hands, instruments availability and the cost) was my first priority. On your request I can send you the copy of this paper;
Russian Journal of Bioorganic Chemistry
November 2002, Volume 28, Issue 6, pp 501-514
The Determination of Total Concentration and Activity of Antioxidants in Foodstuffs
Yu. V. Geletii, G. G. A. Balavoine, O. N. Efimov, V. S. Kulikova,
where you can find a protocol applied for the plant extracts
There are many ways to assess the antioxidant activity and easiest way to evaluate the DPPH or BBC and RP.
You can download these methods of research and of my research
Whatever method is suggested/recommended, make sure it is a published and recognized validated method. ORAC is such a method. Simple doesn't always meaning better. Science is about reproducibility. Getting different results from different labs using exactly the same methods, materials and procedures, the agree, is a step in the right direction.
Following are the most easy and simple method for evaluation of in vitro antioxidant activity of plants..........
1. DDPH
2. Fe++ ion chelating ability
3. Determination of total phenolic content
4. Determination of flavonoid content
You can see the following articles :...
http://www.sciencedirect.com/science/article/pii/S0974694313002387
http://pelagiaresearchlibrary.com/european-journal-of-experimental-biology/vol3-iss1/EJEB-2013-3-1-449-454.pdf
http://www.academicjournals.org/journal/JMPR/article-abstract/E83A3E648091
http://ajp.mums.ac.ir/jufile?c2hvd1BERj0xMDU2Jl9hY3Rpb249c2hvd1BERiZhcnRpY2xlPTEwNTYmX29iPTVjYWVmYTdmZGMwM2RmNjExZDE1YjJiZmI3NTUyODBlJmZpbGVOYW1lPWZ1bGxfdGV4dC5wZGY
In our lab, we usually use DPPH method in order to investigate the antioxidative property of the extracts from plants in vitro. Also, reducing power assay could be use to scan which initial material has ability of antioxidation.
- Badges
- Science topic
- Similar topics
- Biological Science
- Biota
- Eukaryota
- Viridiplantae
- Streptophyta
- Plants