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UPLC/MS based method for quantitative determination of fatty acid composition in Gram-negative and Gram-positive bacteria
Authors:Merli Špitsmeister [Author Vitae] [Author Vitae]  Raivo Vilu [Author Vitae]
Institution:
  • a Competence Centre of Food and Fermentation Technologies, Akadeemia tee 15, 12618, Tallinn, Estonia
  • b Tallinn University of Technology, Department of Food Processing, Ehitajate tee 5, 19086, Tallinn, Estonia
  • c Tallinn University of Technology, Department of Chemistry, Ehitajate tee 5, 19086, Tallinn, Estonia
  • Abstract:Quantitative fatty acid composition of microorganisms at various growth space points is required for understanding membrane associated processes of cells, but the majority of the relevant publications still restrict to the relative compositions. In the current study, a simple and reliable method for quantitative measurement of fatty acid content in bacterial biomass without prior derivatization using ultra performance liquid chromatography-electrospray ionization mass spectrometry was developed. The method was applied for investigating the influence of specific growth rate and pH on the fatty acid profiles of two biotechnologically important microorganisms — Gram-negative bacteria Escherichia coli and Gram-positive bacteria Lactococcus lactis grown in controlled physiological states. It was found that the membranes of slowly growing cells are more rigid and that the fatty acid fraction of the cells of L. lactis diminishes considerably with increasing growth rate.
    Keywords:APCI  Atmospheric pressure chemical ionization  APE  Absolute percent error  CFA  Cyclopropane fatty acids  EI  Electron ionization  ESI-MS  Electrospray ionization mass spectrometry  FAME  Fatty acid methyl esters  FFA  Free fatty acids  HPLC  High performance liquid chromatography  LAB  Lactic acid bacteria  LOD  Limit of detection  LOQ  Limit of quantification  PBS  Phosphate-buffered saline  RSD  Relative standard deviation  SFA  Saturated fatty acids  TIC  Total ion chromatogram  TOF  Time of flight  UFA  Unsaturated fatty acids  UPLC  Ultra performance liquid chromatography
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