Simultaneous Determination of Total Chlorophyll and Total Carotenoid in Detergent Solubilized Chloroplasts

The simultaneous determination of the total carotenoid and the total chloraphyll chloroplast was investigatgd. As the chlorophylls and the carotenoids were found to be solubilized by the cationic detergents, the total carotenoid determinations were carried out by the so-called elimination method, in which the both isolated pigments from spinach leaves were used as the standards and only the carotenoid was determined spectrophotometrically. Namely, cetyldimethyl ammonium chloride was used as the solubilizing agent, and in absorption spectra of the solubilized total chlorophyll aqueous solutions the two wavelengths having the identical optical densities (401 and 443 nm) might be arbitrarily determined. As the first order relationship between the total carotenoid quantities and the differences of these optical densities was able to be established, the carotenoid determinations of the solubilized chloroplasts were also taken place by estimating the optical densities in the two wavalength. The total chlorophyll determinations were carried out by the estimation of the optical densities of the two wavelength (620 and 667 nm). The determination examples of both pigments in some chloroplasts were carried out with these methods.