Temperature-programmed high performance liquid chromatography separation of mono- and divinyl chlorophyll forms from marine phytoplankton

Abstract

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 a5 monomethyl 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.