Precise estimation of chlorophyll a , b and carotenoid content by deconvolution of the absorption spectrum and new simultaneous equations for Chl determination - Archive ouverte HAL
Article Dans Une Revue The Plant Journal Année : 2021

Precise estimation of chlorophyll a , b and carotenoid content by deconvolution of the absorption spectrum and new simultaneous equations for Chl determination

Résumé

The precise determination of photosynthetic pigment content in green organisms, chlorophylls (Chls) andcarotenoids (Cars), is important to investigate many photosynthetic processes such as responses to envi-ronmental fluctuations or to gene mutations, as well as to interpret biochemical and structural resultsobtained on purified membranes and photosynthetic complexes. The most utilized methods for determina-tion by spectrophotometry of Chl content in solution, usually 80% acetone, are based on the use of simulta-neous equations. The advantages are the easiness and speed over chromatography, which also requiresless common equipment. The disadvantage is that issues in sample preparation or in the measurement arenot detectable, which could lead to wrong results. Here we propose a fast, accurate and (almost) error-proof method to measure Chla, Chlband also total Car content in a solution of pigments extracted fromtissue, membranes or purified complexes. The method is based on the fit of the absorption spectrum of theacetone extract using the spectra of purified pigments as references. We show how this method allows amore precise and accurate estimation of pigment content as compared to classical equations, even in incor-rectly prepared acetone solutions. Moreover, the method allows the discovery of artifacts in sample prepa-ration or measurement and thus drastically reduces the risk of mistakes. Examples obtained on purifiedcomplexes are also discussed. Based on newly acquired Chl spectra, we also propose a new set of improvedsimultaneous equations that provide slightly different but more reliable results in comparison with the cur-rently used equations
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Dates et versions

hal-03515372 , version 1 (06-01-2022)

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Marie Chazaux, Christo Schiphorst, Gioele Lazzari, Stefano Caffarri. Precise estimation of chlorophyll a , b and carotenoid content by deconvolution of the absorption spectrum and new simultaneous equations for Chl determination. The Plant Journal, 2021, ⟨10.1111/tpj.15643⟩. ⟨hal-03515372⟩
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