title: High performance liquid chromatographic analysis of phytoplankton pigments using a C16-amide column.
abstract: In this study, a reverse-phase HPLC method incorporating a ternary solvent system was developed to analyze most polar and non-polar chlorophylls and carotenoids present in phytoplankton. The method is based on an RP-C₁₆-Amide column and provided excellent peak resolution of most taxonomically important pigments and an elution profile different than C₈ or C₁₈ columns provide. Analysis of mixed pigment standards, extracts of phytoplankton monocultures, and field samples showed that this method was able to resolve more than sixty pigments, ranging from very polar acidic chlorophylls to the non-polar hydrocarbon carotenes in less than 36 min. This included chlorophylls c₁, c₂ and c₃, divinyl chlorophylls a and b, the carotenoids lutein and zeaxanthin and some recently discovered pigments. The ability of this method to resolve divinyl chl b from monovinyl chl b and divinyl chl a from monovinyl chl a is particularly important for the quantification and identification of the marine cyanobacteria Prochlorococcus spp. in oceanic waters. The described protocol is sensitive and reproducible and can be used to assess the distribution and dynamics of major phytoplankton groups in marine and freshwater ecosystems.
authors.affiliation: Environmental Protection Agency, Office of Research and Development, National Health and Environmental Effects Research Laboratory, Atlantic Ecology Division, 27 Tarzwell Dr., Narragansett, RI 02882, USA. Jayaraman.saro@epa.gov
authors.forename: Saro
authors.initials: S
authors.lastname: Jayaraman
authors.forename: Michael L
authors.initials: ML
authors.lastname: Knuth
authors.forename: Mark
authors.initials: M
authors.lastname: Cantwell
authors.forename: Antelmo
authors.initials: A
authors.lastname: Santos
mesh_terms: D000577:Amides; D002247:Carbon Isotopes / Q000737:chemistry*; D002338:Carotenoids / Q000032:analysis*; D002734:Chlorophyll / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D056148:Chromatography, Reverse-Phase; D010839:Phytoplankton / Q000737:chemistry*; D048929:Prochlorococcus; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014753:Vinyl Compounds
chemical_list: D000577:Amides; D002247:Carbon Isotopes; D014753:Vinyl Compounds; C510432:divinyl chlorophyll a; D002734:Chlorophyll; D002338:Carotenoids