The use of thin-layer chromatography for the separation and identification of phytoplankton pigments

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Abstract
Thin-layer chromatography has been used for the rapid separation of microgram amounts of pigments of marine phytoplankton, both from cultures and from the sea. The separation is carried out on plates coated with silica gel; the chromatogram is developed with a solvent consisting of petroleum ether (b.p. 60–80°), ethyl acetate and diethylamine in the ratio 58:30:12 by volume. The separated individual pigments may be eluted from the plate and characterized by their absorption spectra. The pigments of the following phytoplankton have been examined: Dunaliella primolecta, Phaeodactylum tricornutum, Isochrysis galbana, Dicrateria inornata, Coccolithus huxleyi, and Peridinium trochoidium.IntroductionA knowledge of the component pigments of phytoplankton is of value in the systematic classification of these organisms and in the study of the mechanism of photosynthesis. Very little is known about the amounts of the lesser phyto-plankton pigments associated with the natural phytoplankton crop in the sea, but a considerable amount of work has been carried out on the measurement of chlorophylls a, b and c and of carotenoids for the estimation of the plant bio-mass in the sea. These determinations are usually carried out by modifications of the poly-chromatic spectrophotometric method originally described by Richards with Thompson (1952) and modified by Creitz & Richards (1955) (see also Parsons & Strickland, 1963). While these procedures give a satisfactory measure of chlorophylls a and b, their precision for chlorophyll c and carotenoids is poor; furthermore, no resolution of carotenes from xanthophylls can be attained; nor can any indication of the presence of unusual pigments be obtained.