Separation of chlorophylls and carotenoids from marine phytoplankton:a new HPLC method using a reversed phase C8 column and pyridine-containing mobile phases

TLDR

The study introduces a reversed‑phase C8 HPLC method with pyridine‑containing mobile phases that achieves simultaneous separation of chlorophylls and carotenoids, a capability previously limited to polymeric C18 columns. Using a binary gradient, the method is compatible with both low‑ and high‑pressure mixing instruments and was validated across three distinct HPLC systems. It resolves monovinyl/divinyl chlorophyll pairs—including DV chlorophyll a versus chlorophyll b—and separates taxon‑specific carotenoids from eight algal classes, requiring only minor gradient adjustments for different equipment. © 2000 Inter‑Research; keywords: HPLC pigment analysis, phytoplankton pigments, C8 column, pyridine‑containing mobile phases; published in Marine Ecology Progress Series vol.

Abstract

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 195:29-45 (2000) - doi:10.3354/meps195029 Separation of chlorophylls and carotenoids from marine phytoplankton: a new HPLC method using a reversed phase C8 column and pyridine-containing mobile phases Manuel Zapata1,*, Francisco Rodríguez1, José L. Garrido2 1Centro de Investigacións Mariñas, Consellería de Pesca, Xunta de Galicia, Apdo.13, 36620 Vilanova de Arousa, Spain 2Instituto de Investigacións Mariñas de Vigo (CSIC), Eduardo Cabello 6, 36208 Vigo, Spain *E-mail: [e-mail] ABSTRACT: A high-performance liquid chromatographic (HPLC) method based on a reversed-phase C8 column and pyridine-containing mobile phases was developed for the simultaneous separation of chlorophylls and carotenoids. The method is selective enough to resolve monovinyl (MV) and divinyl (DV) pairs of polar chlorophylls and DV chlorophyll a (chl a) (the marker pigment for the prokaryote Prochlorococcus marinus) from chl a (the MV analogue). Only the pair DV chl a/chl b was not resolved. This resolution capability for chlorophylls was only previously achieved using polymeric C18 columns in combination with ammonium acetate or pyridine-containing mobile phases. The proposed method also allows the separation of taxon-specific carotenoids belonging to 8 algal classes, including some critical pigment pairs for previous HPLC methods using C18 columns. The method employs a binary gradient, so it can be used with both low-pressure and high-pressure mixing instruments. Method transferability was tested using 3 HPLC systems. Only a slight adjustment of gradient profile was required to obtain similar results with HPLC equipment having different dwell volumes. The selectivity of the method towards some recently discovered chlorophyll and carotenoid pigments makes it especially suitable for studying not only field samples, but also for re-examining the pigment composition of different algal classes. KEY WORDS: HPLC pigment analysis · Phytoplankton pigments · C8 column · Pyridine-containing mobile phases · Chemotaxonomy Full text in pdf format PreviousNextExport citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 195. Publication date: March 31, 2000 Print ISSN:0171-8630; Online ISSN:1616-1599 Copyright © 2000 Inter-Research.