Formulae for Determination of Chlorophyllous Pigments Extracted with N,N-Dimethylformamide

Abstract

The extraction of chlorophylls in higher plant tissue using N,N-dimethylformamide expedites the process and enables the determination of small samples with low pigment level.

Absorption spectra of Chl a, Chl b, and Pchl and of their acidified derivatives, the phaeophytins, were recorded. Conversion of Chl b to its corresponding acidified product occurs much more slowly than that of Chl a and Pchl. When acidified, Pchl differs from Chl a and Chl b by the disappearance of the red band in the absorption spectrum. Specific extinction coefficients were determined and formulae for quantitative determination of pigments concentrations were developed. When concentrations of pigments are low, as in etiolated plant material, the absorption spectra of the chlorophylls can be distorted due to the presence of other substances simultaneously extracted; formulae for pigment determination under such circumstances were also derived.