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Evaluation of various chromatographic approaches for the retention of hydrophilic compounds and MS compatibility

Published in Journal of Separation Science. 2013, vol. 36, p. 3141-3151
Abstract The goal of this study was to compare the performance of three separation techniques for the analysis of 57 hydrophilic compounds. RPLC, hydrophilic interaction liquid chromatography (HILIC) and subcritical fluid chromatography (SFC) were tested. The comparison was based on the retention, selectivity, peak shape (asymmetry and peak width) and MS sensitivity. As expected, RPLC had some obvious limitations for such classes of compounds, and on average the %ACN required to elute these hydrophilic substances was 4, 7, and 11% ACN at pH 3, 6, and 9, respectively. However, a hybrid polar-embedded C18 phase with an appropriate mobile phase could represent a viable strategy for hydrophilic basic compounds with log D greater than -2 on average. HILIC and SFC were found to be more appropriate for analyzing a large majority of these hydrophilic analytes (~60 and 70% of compounds eluted during the gradient in HILIC and SFC), while maintaining good MS sensitivity. Finally, this work demonstrated the complementarity of the three analytical techniques and showed that the selection of a suitable strategy should mostly be based on physicochemical properties of the analytes (pKa, log D, H-bonding capability, etc.).
Keywords HILICMS sensitivityPolar substancesRPLCSubcritical fluid chromatography
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Other version: http://doi.wiley.com/10.1002/jssc.201300567
Research group Sciences analytiques
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PERIAT, Aurélie Claudine, GRAND-GUILLAUME-PERRENOUD, Alexandre, GUILLARME, Davy. Evaluation of various chromatographic approaches for the retention of hydrophilic compounds and MS compatibility. In: Journal of Separation Science, 2013, vol. 36, p. 3141-3151. https://archive-ouverte.unige.ch/unige:32773

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Deposited on : 2014-01-06

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