Hello every one,
I extract chlorophyll , and I read the absorbance in 480, 663 and 645.
I need to estimate the quantity of both chlorophyll and cartonoid , I need help please. Could you ?
Thank you
Hello Hafida
1) Considering that you have to measure the absorbance at these particular wavelengths (480, 645 and 663 nm) I think that you can use the equations presented in the attached paper 1 (pages 206-207).
2) Substitutionally, you can see the following protocol based on the attached paper 2 which uses different equations and wavelengths related to different solvents:
The extraction procedure of the photosynthetic pigments is carried out under dim light and low temperatute (4oC). The plant leaf material (0.5 gr is enough) is ground in porcelain mortar using 100% acetone (or other suitable solvent, see paper 2). The resulting suspension is centrifuged at 4oC at 2000 g for 10 min. Chlorophyll and carotenoids contents are determined in acetone supernatants (or in appropriate dilutions) according to the method of Lichtenthaler (paper 2). At page 366 you can find acetone or other solvents' equations for chlorophyll a, b and carotenoids spectrophotometric quantification.
Hello Hafida
1) Considering that you have to measure the absorbance at these particular wavelengths (480, 645 and 663 nm) I think that you can use the equations presented in the attached paper 1 (pages 206-207).
2) Substitutionally, you can see the following protocol based on the attached paper 2 which uses different equations and wavelengths related to different solvents:
The extraction procedure of the photosynthetic pigments is carried out under dim light and low temperatute (4oC). The plant leaf material (0.5 gr is enough) is ground in porcelain mortar using 100% acetone (or other suitable solvent, see paper 2). The resulting suspension is centrifuged at 4oC at 2000 g for 10 min. Chlorophyll and carotenoids contents are determined in acetone supernatants (or in appropriate dilutions) according to the method of Lichtenthaler (paper 2). At page 366 you can find acetone or other solvents' equations for chlorophyll a, b and carotenoids spectrophotometric quantification.
Dear Hafida Baoune,
Please you can read this research for estimation the quantity of both chlorophyll and cartonoid
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1679-87592014000100006
Best regards
Chlorophyll quantification can be done using Arnons formula (1949).
Arnon (1949) gave the equations for extraction with 80% acetone and absorbances read at 645nm and 663 nm as
Chlorophyll a (μg/ml) = 12.7 (A663) - 2.69 (A645)
Chlorophyll b (μg/ml) = 22.9 (A645) - 4.68 (A663)
Thank you so much for all.
I make the calculations and I am glad , I find a good results.
Thank you George Zervoudakis, I use the paper 1 for my calculations.
Thank you Kais Kassim , the paper that you offer me , it is really interesting.
Thank you Liza, I used this formula.
With my kind regards for you all.
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