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HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY OF PHYTOPLANKTON PIGMENTS USING A POLYMERIC REVERSED-PHASE C18 COLUMN1
Authors:Laurie Van Heukelem  Alan J. Lewitus  Todd M. Kana  Neal E. Craft
Abstract:A high-performance liquid chromatographic (HPLC) method is described that allows improved resolution of several chemotaxonomically significant phytoplankton pigments. The protocol, which employs two pumps and a modified Mantoura and Llewellyn (1983) solvent system, can be easily adapted for many HPLC systems currently in use. The most unique aspect of the method is the use of a polymeric C18 reversed phase HPLC column (VYDAC 201TP). In comparison to the monomeric C18 columns typically used in the characterization of phytoplankton pigments, polymeric C18 columns offer superior selectivity for structurally similar compounds. The protocol was evaluated for the ability to resolve most of the phytoplankton pigments of diagnostic importance using algal cultures from nine classes. Pigment pairs that were resolved by the method include a) lutein and zeaxanthin, b) neoxanthin and 19′-hexanoyloxyfucoxanthin, and c) α-carotene and β-carotene, and partial resolution of chlorophyll c1 and chlorophyll c2.
Keywords:carotenoids  chlorophyll c  high-performance liquid chromatography (HPLC)  lutein  phytoplankton  pigments  polymeric  reversed-phase HPLC  zeaxanthin
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