Skip to Content
Merck
  • Comprehensive methodology for Staphylococcus aureus lipidomics by liquid chromatography and quadrupole time-of-flight mass spectrometry.

Comprehensive methodology for Staphylococcus aureus lipidomics by liquid chromatography and quadrupole time-of-flight mass spectrometry.

Journal of chromatography. A (2014-08-28)
Weronika Hewelt-Belka, Joanna Nakonieczna, Mariusz Belka, Tomasz Bączek, Jacek Namieśnik, Agata Kot-Wasik
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.

MATERIALS
Product Number
Brand
Product Description

Supelco
2-Propanol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Methanol-12C, 99.95 atom % 12C
Supelco
2-Propanol, analytical standard
Sigma-Aldrich
2-Propanol, BioUltra, for molecular biology, ≥99.5% (GC)
Sigma-Aldrich
2-Propanol, anhydrous, 99.5%
Sigma-Aldrich
Hexane, anhydrous, 95%
Supelco
Hexane, analytical standard
Sigma-Aldrich
2-Propanol, for molecular biology, BioReagent, ≥99.5%
Sigma-Aldrich
2-Propanol, electronic grade, 99.999% trace metals basis
Sigma-Aldrich
2-Propanol, puriss. p.a., ACS reagent, ≥99.8% (GC)
Sigma-Aldrich
2-Propanol, suitable for HPLC, 99.9%
Sigma-Aldrich
Hexane, ReagentPlus®, ≥99%
Sigma-Aldrich
2-Propanol, HPLC Plus, for HPLC, GC, and residue analysis, 99.9%
USP
2-Propanol, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Hexane, suitable for HPLC, ≥95%
Sigma-Aldrich
Hexane, suitable for HPLC, ≥97.0% (GC)
Sigma-Aldrich
2-Propanol, ACS reagent, ≥99.5%
Sigma-Aldrich
Hexane, HPLC Plus, for HPLC, GC, and residue analysis, ≥95%
Sigma-Aldrich
Methanol, NMR reference standard
Sigma-Aldrich
Chloroform, anhydrous, ≥99%, contains 0.5-1.0% ethanol as stabilizer
Sigma-Aldrich
Isopropyl alcohol, ≥99.7%, FCC, FG
Sigma-Aldrich
Methanol, anhydrous, 99.8%
Supelco
Methanol, analytical standard
Sigma-Aldrich
Ammonium formate, BioUltra, ≥99.0% (calc. based on dry substance, NT)
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Supelco
Ammonium formate, eluent additive for LC-MS, LiChropur, ≥99.0%
Sigma-Aldrich
Ammonium formate, ≥99.995% trace metals basis
Sigma-Aldrich
Methanol, BioReagent, ≥99.93%
Sigma-Aldrich
Methanol, HPLC Plus, ≥99.9%
Sigma-Aldrich
Chloroform, suitable for HPLC, ≥99.8%, amylene stabilized