title: Analysis of non-derivatised bacteriohopanepolyols by ultrahigh-performance liquid chromatography/tandem mass spectrometry.
abstract: BACKGROUND
  Traditional investigation of bacteriohopanepolyols (BHPs) has relied on derivatisation by acetylation prior to gas chromatography/mass spectrometry (GC/MS) or liquid chromatography/MS (LC/MS) analysis. Here, modern chromatographic techniques (ultrahigh-performance liquid chromatography (UPLC)) and new column chemistries were tested to develop a method for BHP analysis without the need for derivatisation.
  
  
  METHODS
  Bacterial culture and sedimentary lipid extracts were analysed using a Waters Acquity Xevo TQ-S triple quadrupole mass spectrometer in positive ion atmospheric pressure chemical ionisation (APCI) mode. Waters BEH C18 and ACE Excel C18 were the central columns evaluated using a binary solvent gradient with 0.1% formic acid in the polar solvent phase in order to optimise performance and selectivity.
  
  
  RESULTS
  Non-amine BHPs and adenosylhopane showed similar performance on each C18 column; however, BHP-containing terminal amines were only identified eluting from the ultra-inert ACE Excel C18 column. APCI-MS/MS product ion scans revealed significant differences in fragmentation pathways from previous methods for acetylated compounds. The product ions used for targeted multiple reaction monitoring (MRM) are summarised.
  
  
  CONCLUSIONS
  UPLC/MS/MS analysis using an ACE Excel C18 column produced superior separation for amine-containing BHPs and reduced run times from 60 to 9 min compared with previous methods. Unexpected variations in fragmentation pathways between structural subgroups must be taken into account when optimising MRM transitions for future quantitative studies. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: School of Civil Engineering and Geosciences, Newcastle University, Newcastle upon Tyne, NE1 7RU, UK.
authors.forename: Helen M
authors.initials: HM
authors.lastname: Talbot
authors.affiliation: School of Civil Engineering and Geosciences, Newcastle University, Newcastle upon Tyne, NE1 7RU, UK.|Newcastle University Protein and Proteome Analysis (NUPPA), Devonshire Building, Newcastle University, Newcastle upon Tyne, NE1 7RU, UK.
authors.forename: Frances R
authors.initials: FR
authors.lastname: Sidgwick
authors.affiliation: School of Civil Engineering and Geosciences, Newcastle University, Newcastle upon Tyne, NE1 7RU, UK.
authors.forename: Juliane
authors.initials: J
authors.lastname: Bischoff
authors.affiliation: School of Civil Engineering and Geosciences, Newcastle University, Newcastle upon Tyne, NE1 7RU, UK.
authors.forename: Kate A
authors.initials: KA
authors.lastname: Osborne
authors.affiliation: School of Civil Engineering and Geosciences, Newcastle University, Newcastle upon Tyne, NE1 7RU, UK.
authors.forename: Darci
authors.initials: D
authors.lastname: Rush
authors.affiliation: School of Civil Engineering and Geosciences, Newcastle University, Newcastle upon Tyne, NE1 7RU, UK.
authors.forename: Angela
authors.initials: A
authors.lastname: Sherry
authors.affiliation: School of Civil Engineering and Geosciences, Newcastle University, Newcastle upon Tyne, NE1 7RU, UK.
authors.forename: Charlotte L
authors.initials: CL
authors.lastname: Spencer-Jones
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008563:Membrane Lipids / Q000737:chemistry* / Q000378:metabolism; D020619:Methylococcus capsulatus / Q000737:chemistry* / Q000378:metabolism; D020602:Methylosinus trichosporium / Q000737:chemistry* / Q000378:metabolism; D015394:Molecular Structure; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D008563:Membrane Lipids