MDA TBARS Malondialdehyde
I want to use different concentration of plant extract and the measure the MDA content from final product, but plant extract has its natural color, so it disturb the final color of reaction mixture and cause variation in OD. To solve this problem, Can we take OD of reaction mixture before TBARS solution addition and heating and minus this OD from the final OD of reaction mixture?
Is this correct way to remove the difference of natural color of plant extracts, or there is any other suitable way?
Thanks
I think that you can solve your problem if you prepare a sample blank and subtract its OD from the corresponding sample OD.
The sample blank must be prepared and measured under exactly the same conditions (quantity of extract, incubation, heating etc.) with the corresponding sample, with only one difference: the reaction solution added to the extract sample must not contain thiobarbituric acid (TBA). Thus, your sample blank will measure only the non-specific absorbance including the natural colour of the extract.
I think that you can solve your problem if you prepare a sample blank and subtract its OD from the corresponding sample OD.
The sample blank must be prepared and measured under exactly the same conditions (quantity of extract, incubation, heating etc.) with the corresponding sample, with only one difference: the reaction solution added to the extract sample must not contain thiobarbituric acid (TBA). Thus, your sample blank will measure only the non-specific absorbance including the natural colour of the extract.
If you intend to evaluate the antioxidant activity in a range of lipid peroxidation systems using rat liver microsomes, red blood cells and other in vitro assays to determine their ability to scavenge free radicals, should be carried out appropriate controls to discard any possible interference with the assay (Schinella et al, Life Sci. 2002)
For example, if evaluated Lipid peroxidation induced by Fe (II) / ascorbate, checks are necessary, such as: 1) the extract in the presence of microsomes and absence Fe (II) / ascorbate, and 2) the extract in the presence of Fe (II) / ascorbate and absence of microsomes (It is important to note that sugars present in the extract in the presence of a prooxidant system could generate MDA).
Schinella GR, Tournier HA, Prieto JM, Mordujovich de Buschiazzo P, Ríos JL. Antioxidant activity of anti-inflammatory plant extracts. Life Sci. 2002; 70(9):1023-33.
Completely agreed with George Z.
Alternatively, decolorize the test extract using activated charcoal pass under multi-stage vacuum filtration. A quick check of antioxidant activity (DPPH test) (pre and post filtration) can help optimize the no. of cycle pass, and ensures maximum retention of activity.
Good luck!
thank you very much all of you for your valuable time and considerations
Dear add ur blank solution in THE complete preparation of set wit plant extract only and substrate it from final odq
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