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Divinyl chlorophyll a (a2) and divinyl chlorophyll b (b2) are chemotaxonomic marker pigments for the marine prochlorophytes, and can be used to study their distribution in marine samples. In this paper we report a baseline resolution of mono- and divinyl forms of chlorophylls a and b employing polymeric ODS stationary phases at sub-ambient temperatures. The simultaneous resolution of mono- and divinyl forms of chlorophylls a and b, chlorophylls c1, c2, c3, Mg 3,8-divinylphaeoporphyrin a5monomethyl ester (MgDVP), and phytol-substituted chlorophylls c was achieved when a temperature step gradient was employed during the analysis. An example is given of the utility of the protocol in oceanic field samples.

Photosynthetic pigments from chromophyte marine algae were analyzed using three different commercial columns with ODS stationary phases of different degrees of polymerization. No separation of chlorophylls c1 and c2 could be achieved using monomeric or intermediate stationary phases, even at subambient temperatures. As with other planar compounds, the polymeric stationary phase showed the highest selectivity for chlorophyll c pigments, allowing the simultaneous separation of chlorophylls c3, c1 and c2 and other pigments occurring in chromophyte algae by using a binary gradient starting with a highly buffered mobile phase.