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Title

Applicability of supercritical fluid chromatography–mass spectrometry to metabolomics. I – Optimization of separation conditions for the simultaneous analysis of hydrophilic and lipophilic substances

Authors
Farrell, William P.
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Published in Journal of Chromatography. A. 2018, vol. 1562, p. 96-107
Abstract The aim of this study was to evaluate the suitability of SFC-MS for the analysis of a wide range of compounds including lipophilic and highly hydrophilic substances (log P values comprised between − 6 and 11), for its potential application toward human metabolomics. For this purpose, a generic unified chromatography gradient from 2 to 100% organic modifier in CO 2 was systematically applied. In terms of chemistry, the best stationary phases for this application were found to be the Agilent Poroshell HILIC (bare silica) and Macherey-Nagel Nucleoshell HILIC (silica bonded with a zwitterionic ligand). To avoid system overpressure at very high organic modifier proportion, columns of 100×3mm I.D. packed with sub-3m superficially porous particles were selected. In terms of organic modifier, a mixture of 95% MeOH and 5% water was selected, with 50mM ammonium formate and 1 mM ammonium fluoride, to afford good solubility of analytes in the mobile phase, limited retention for the most hydrophilic metabolites and suitable peak shapes of ionizable species. A sample diluent containing 50%ACN/50% water was employed as injection solvent. These conditions were applied to a representative set of metabolites belonging to nucleosides, nucleotides, small organic acids, small bases, sulfated/sulfonated metabolites, poly-alcohols, lipid related substances, quaternary ammonium metabolites, phosphate-based substances, carbohydrates and amino acids. Among all these metabolites, 65% of the compounds were adequately analyzed with excellent peak shape, 23% provided distorted peak shapes, while only 12% were not detected (mostly métabolites having several phosphate or several carboxylic acid groups).
Keywords Supercritical fluid chromatographyPolar compoundsMetabolomicsSFC-MS
Identifiers
PMID: 29861304
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Structures
Research group Sciences analytiques
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DESFONTAINE, Vincent et al. Applicability of supercritical fluid chromatography–mass spectrometry to metabolomics. I – Optimization of separation conditions for the simultaneous analysis of hydrophilic and lipophilic substances. In: Journal of Chromatography. A, 2018, vol. 1562, p. 96-107. https://archive-ouverte.unige.ch/unige:106684

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Deposited on : 2018-07-25

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