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Steevensz, AJ,MacKinnon, SL,Hankinson, R,Craft, C,Connan, S,Stengel, DB,Melanson, JE
Phytochemical Analysis
Profiling Phlorotannins in Brown Macroalgae by Liquid Chromatography-High Resolution Mass Spectrometry
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high resolution mass spectrometry hydrophilic interaction chromatography brown seaweed phlorotannins polyphenols HYDROPHILIC-INTERACTION CHROMATOGRAPHY ALGA FUCUS-VESICULOSUS ECKLONIA-KUROME ANTIOXIDANT PHAEOPHYCEAE POLYPHENOLS SEPARATION MODE CAVA WALL
Introduction Phlorotannins, phenolic compounds produced exclusively by Phaeophyceae (brown algae), have recently been associated with a wide variety of beneficial bioactivities. Several studies have measured the total phenolic content in extracts from various species, but little characterisation of individual phlorotannin components has been demonstrated. Objective The purpose of this study was to develop a liquid chromatographymass spectrometry (LC-MS) based method for rapid profiling of phlorotannins in brown algae. Methodology Phlorotannin-enriched extracts from five phaeophyceaen species were analysed by ultrahigh-pressure liquid chromatography (UHPLC) operating in hydrophilic interaction liquid chromatography (HILIC) mode combined with high resolution mass spectrometry (HRMS). The method was optimised using an extract of Fucus vesiculosus; separation was achieved in less than 15?min. The basic mobile phase enhanced negative-ion electrospray ionisation (ESI), and generated multiply charged ions that allowed detection of high molecular weight phlorotannins. Results The phlorotannin profiles of Pelvetia canaliculata, Fucus spiralis, F. vesiculosus, Ascophyllum nodosum and Saccharina longicruris differed significantly. Fucus vesiculosus yielded a high abundance of low molecular weight (< 1200?Da) phlorotannins, while P. canaliculata exhibited a more evenly distributed profile, with moderate degrees of polymerisation ranging from 3 to 49. HRMS enabled the identification of phlorotannins with masses up to 6000?Da using a combination of accurate mass and 13?C isotopic patterns. Conclusion The UHPLC-HRMS method described was successful in rapidly profiling phlorotannins in brown seaweeds based on their degree of polymerisation. HILIC was demonstrated to be an effective separation mode, particularly for low molecular weight phlorotannins. Copyright (c) 2012 John Wiley & Sons, Ltd.
DOI 10.1002/pca.2354
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