Can someone describe me a simple protocol to determinate the total phenols in plants in order to study the effect of salt stress in it? Is the Folin Ciocalteu method is the simplest? Thanks.
Yes you can use Folin Ciocalteu method to estimate total phenol content in plants to study the effect of salt stress. The method is simple and the paper attached below can help you.
Folin Ciocalteu method is a good one for total phenolics - just see that you don't get a very dark color
I used this method. I hope it can help you
Total phenolic content was determined by a method based on Folin-ciocalteu assay with slight modification. Ground dry bean sample (0.5g) was treated with Methanol for 30 min. clear supernatant was centrifuged at 10,000 rpm for 10 min at 25⁰c. Sample extract (300 µl), distilled water (200 µl), Folin-ciocalteu reagents (2.5 ml), and 7% Naco3 (2 ml) were mixed and incubated in darkness for 1.5 hr at room temperature. The absorbance reading was taken at 760 nm using a UV-visible spectrophotometer. The results were expressed as Gallic acid equivalents (GAE).
I agree. The simplest is the Folin-Ciocalteu assay and you can use whatever phenolic you have at hand as standard equivalent: phenolic, caffeic or cumaric acid besides gallic.
Malick and Singh(1980) is a best method used in the plant stress physiology works extract 500 mg fresh tissue with10 ml of 80 % etanol, then centrifuge homogenate @ 10000 rpm for 20 min, allow to evaporate the supernatant till dry, followed by dissolve the residue in 5ml of distilled water n you can use that as extract.
For estimation 2ml extract/.5ml Folin-Ciocalteu reagent, after 3 min, add 1ml 20% Na2CO3 , mix well. Keep in boiling water for exactly one minute, after cooling read the absorbance @ 650 nm (Blank is water n reagent)
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