title: Comprehensive methodology for Staphylococcus aureus lipidomics by liquid chromatography and quadrupole time-of-flight mass spectrometry.
abstract: Staphylococcus aureus is a common pathogen known to cause relatively minor infections as well as severe disorders in humans. Although there is fair amount of published data concerning various aspects of its biology, epidemiology, genetics, etc., there is still a scarce amount of data presenting reliable and thorough investigations regarding high-throughput analysis of total S. aureus lipid content. Therefore, the aim of this study was to develop an analytical method that in combination with advanced chemometric tools enables comprehensive lipidomic analysis of S. aureus cells. The newly developed method uses high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) to directly examine extracted lipids and further identify them within a newly developed novel S. aureus lipids database. High coverage of the S. aureus lipidome was obtained by simultaneous bacterial cell lysis and liquid-liquid extraction. The combination of three techniques enabled the analysis of the major membrane lipid classes of S. aureus: separation of the lipid extract in reversed-phase mode, Q-TOF-MS detection in positive ion mode, and lipid database mining. The developed lipidomic approach is also a powerful tool that allows one to assess the lipid content of S. aureus cells in a comparative manner between strains characterized by different phenotypic features as exemplified here by their various sensitivities toward antibiotics.
authors.affiliation: Department of Analytical Chemistry, Chemical Faculty, Gdańsk University of Technology, G. Narutowicza 11/12, 80-233 Gdańsk, Poland. Electronic address: weronika.hb@gmail.com.
authors.forename: Weronika
authors.initials: W
authors.lastname: Hewelt-Belka
authors.affiliation: Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, Kładki 24, 80-822 Gdańsk, Poland. Electronic address: joanna.nakonieczna@biotech.ug.edu.pl.
authors.forename: Joanna
authors.initials: J
authors.lastname: Nakonieczna
authors.affiliation: Department of Pharmaceutical Chemistry, Medical University of Gdańsk, Hallera 107, 80-416 Gdańsk, Poland. Electronic address: mariusz.belka@gmail.com.
authors.forename: Mariusz
authors.initials: M
authors.lastname: Belka
authors.affiliation: Department of Pharmaceutical Chemistry, Medical University of Gdańsk, Hallera 107, 80-416 Gdańsk, Poland. Electronic address: tbaczek@gumed.edu.pl.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Bączek
authors.affiliation: Department of Analytical Chemistry, Chemical Faculty, Gdańsk University of Technology, G. Narutowicza 11/12, 80-233 Gdańsk, Poland. Electronic address: jacek.namiesnik@pg.gda.pl.
authors.forename: Jacek
authors.initials: J
authors.lastname: Namieśnik
authors.affiliation: Department of Analytical Chemistry, Chemical Faculty, Gdańsk University of Technology, G. Narutowicza 11/12, 80-233 Gdańsk, Poland. Electronic address: agawasik@pg.gda.pl.
authors.forename: Agata
authors.initials: A
authors.lastname: Kot-Wasik
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019295:Computational Biology / Q000379:methods*; D050356:Lipid Metabolism; D008055:Lipids / Q000032:analysis* / Q000737:chemistry; D059625:Liquid-Liquid Extraction / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods*; D013211:Staphylococcus aureus / Q000737:chemistry* / Q000378:metabolism
chemical_list: D008055:Lipids
keywords: Chemometric analysis; Comparative non-targeted lipidomics; Lipid database; Liquid chromatography–mass spectrometry; Staphylococcus aureus lipids