I recently needed to quantify chlorophylls and found that most
researchers continue to use the original method of Arnon (Arnon, D.I.
(1949) Plant Physiol. 24, 115). However, subsequent studies indicate
inaccuracies in his extinction coefficients.
I decided to use the highly-cited method of Porra et al. (Biochimica et
Biophysica Acta (BBA) - Bioenergetics. Volume 975, Issue 3, August 1989,
Pages 384-394). The equations also appear in a review by Porra
(PHOTOSYNTHESIS RESEARCH Volume: 73 Issue: 1-3 Pages: 149-156).
They present updated equations for quantifying chlorophyll a, b, and a +
b in three solvents: buffered aqueous 80% acetone, DMF, and methanol.
I chose 80% acetone.
Postdoctoral Research Associate
arab-gen-request from oat.bio.indiana.edu wrote:
>> Message: 2
> Date: Mon, 3 Mar 2008 15:06:35 -0600
> From: "Siefers, Nicholas A." <nsiefers from ou.edu>
> Subject: [Arabidopsis] Quantification of total chlorophyll;
> chlorophyll a and b
> To: "arab-gen from net.bio.net" <arab-gen from magpie.bio.indiana.edu>
> <ADEA556CB6893A47AB325BC6EC28F7BFEE55417AF1 from XMAIL4.sooner.net.ou.edu>
> Content-Type: text/plain; charset="iso-8859-1"
>> Dear Arab-gen,
>> We are looking for alternative methods of quantifying total chlorophyll and/or chlorphyll a and b. Presently we are using the dimethyl formamide methods as desribed in the two papers:
>> Moran, 1982, Plant Phys, 69:1376
>> Inskeep 1985, Plant Phys, 77:483
>> If your lab is using an alternative method for quantification of chlorophyll please notify me via email at:
>nsiefers from ou.edu<mailto:nsiefers from ou.edu>
>> thanks in advance,
> Nicholas Siefers
> Holt Lab
> University of Oklahoma