I want to calculate spectrophotometrically the concentration of Chl a and b after extraction with acetone 90%.
Can I use the equation from Jeffrey and Humphrey (1975), even if green algae do not have chlorophyll c?
Moreover, does anyone know the same equations if I use ethanol as solvent for pigment extraction?
There are several methods for the determination of chlorophyll pigments using acetone90% , and it is easy to calculate chl.a and chl.b using published equations of Holden,1965
Please, use the attached paper about chlorophyll extraction from green algae.
Dear Lorenza,
Depending on alga species, different extraction methods ensure different extraction yields. The ethanolic extraction is rather questionable, in some species, it extracts only about 10 % of chlorophylls. I would suggest you the hot methanol method. True, methanol is dangerous but you certainly can organize your work considering all safety regulations. I attach our earlier publication about the ethanol vs. methanol question. You can use the equations and references of this paper, too.
Best greetings and good luck to your work,
Bela
Consult the Book by T.W.Goodwin Chemistry and Biochemistry of Plant Pigments. Volume I .Academic Press.
lorenza ferro! The acetone method you have used for your extraction, or if you use methanol Porra's method for pigment calculation is best. find the attached paper.
I used acetone 100% and EtOH 95% and they both worked well, but I had first to disrupt cell walls through a homogenization step. The results were comparable and the pellet was quite white after 1 or 2 steps extractions.
Chlorophyll was determined according to Wintermans and De Mots (1965) after extraction in 96% (v/v) ethanol.
http://cse.naro.affrc.go.jp/suzuki/Chlorophyll.html
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