title: The Role of Fluorinated Alcohols as Mobile Phase Modifiers for LC-MS Analysis of Oligonucleotides.
abstract: Hexafluoroisopropanol (HFIP) has been widely used as an acidic modifier for mobile phases for liquid chromatography-mass spectrometry (LC-MS) analysis of oligonucleotides ever since the first report of its use for this purpose. This is not surprising, considering the exceptional performance of HFIP compared with carboxylic acids, which cause significant MS signal suppression in electrospray ionization. However, we have found that other fluorinated alcohols can also be utilized for mobile phase preparation and the choice of optimal fluorinated alcohol is determined by the ion-pairing (IP) agent. Although HFIP is a very good choice to be used alongside less hydrophobic IP agents, other fluorinated alcohols such as 1,1,1,3,3,3-hexafluoro-2-methyl-2-propanol (HFMIP) can significantly outperform HFIP when used with more hydrophobic IP agents. We also found that more acidic fluorinated alcohols assist with the transfer of oligonucleotides with secondary structure (e.g., folded strands and hairpins) into the gas phase. Graphical Abstract ᅟ.
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, The University of Georgia College of Pharmacy, 250 W. Green Street, Athens, GA, 30602-2352, USA.
authors.forename: Babak
authors.initials: B
authors.lastname: Basiri
authors.affiliation: ProQR Therapeutics NV, Zernikedreef 9, 2333 CK, Leiden, The Netherlands.
authors.forename: Hilde
authors.initials: H
authors.lastname: van Hattum
authors.affiliation: ProQR Therapeutics NV, Zernikedreef 9, 2333 CK, Leiden, The Netherlands.
authors.forename: William D
authors.initials: WD
authors.lastname: van Dongen
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, The University of Georgia College of Pharmacy, 250 W. Green Street, Athens, GA, 30602-2352, USA.
authors.forename: Mandi M
authors.initials: MM
authors.lastname: Murph
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, The University of Georgia College of Pharmacy, 250 W. Green Street, Athens, GA, 30602-2352, USA. bartlett@rx.uga.edu.
authors.forename: Michael G
authors.initials: MG
authors.lastname: Bartlett
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D004247:DNA / Q000737:chemistry; D054879:Halogenation; D057927:Hydrophobic and Hydrophilic Interactions; D007202:Indicators and Reagents; D008745:Methylation; D009841:Oligonucleotides / Q000737:chemistry*; D020005:Propanols / Q000737:chemistry*; D012313:RNA / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D006845:Hydrocarbons, Fluorinated
chemical_list: D004247:DNA; D007202:Indicators and Reagents; D009841:Oligonucleotides; D020005:Propanols; D012313:RNA; C001337:hexafluoroisopropanol; D006845:Hydrocarbons, Fluorinated
keywords: Electrospray ionization; Fluorinated alcohols; LC-MS; Oligonucleotides