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Chlorophyll content, pigment extraction

Posted by dianaxvb (My Page) on
Mon, Jun 29, 09 at 15:09

Hi everyone
I have been looking for different methods to measure leaf chlorophyll content. And I have found a lot of different approaches: 1. using different solvents (acetone; dimethyl sulphoxide (DMSO); Methanol; ethanol); 2. by griding the samples with the solvents; making a powder out of the samples and then mix it with the solvent; or just making the extraction by leaving the sample in the solvent in a water bath at 65 celcius degrees; 3. using centrifuges or not; 4. using High Performance Liquid Chromatography (HPLC); 5. measuring with spectrophotometers; 6. using Arnon's (1949) equations for the chlorophyll content; or Porra's (1989) equations.
Does anyone have a recommendation about which methods is better?
Can somebody give me a reference that can give me a light about which approach is more accurate or reliable?
Thanks a lot for your help!

Follow-Up Postings:

RE: Chlorophyll content, pigment extraction

Better? I dunno myself, but I do know that, going through college, we would smash up leaves, let them soak in acetone for a week in a refrigerator covered by aluminum foil, then measure them with spectrophotometers. Relatively painless and we got good results.

RE: Chlorophyll content, pigment extraction

I'd go with Terpguy on the extraction business. The secret is low temps. I have used acid-free methanol at ca. 4 degrees C for at least 48 hrs. Then again, which chlorophyll are you trying to determine? Also, very quick drying of the plant tissue and grinding to a fine powder are both good policies. Sounds as tho you are pursuing an interesting project!!!

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