title: Isotope-labeled differential profiling of metabolites using N-benzoyloxysuccinimide derivatization coupled to liquid chromatography/high-resolution tandem mass spectrometry.
abstract: BACKGROUND
  An isotopic labeling strategy based on derivatizing amine-containing metabolites has been developed using light ((12) C6 ) and heavy ((13) C6 ) N-benzoyloxysuccinimide reagents for semi-targeted metabolomic applications.
  
  
  METHODS
  Differentially labeled samples were combined and analyzed simultaneously by liquid chromatography/high-resolution tandem mass spectrometry (LC/HR-MS/MS) to compare relative amounts of amine-containing metabolites. The selectivity of the reaction was determined with model metabolites and was shown to also be applicable to thiol and phenol moieties. The potential for relative quantitation was evaluated in cell extracts and the method was then applied to quantify metabolic perturbations occurring in human cultured cells under normal vs. oxidative stress conditions.
  
  
  RESULTS
  A total of 279 derivatized features were detected in HL60 cell extracts, 77 of which yielded significant concentration changes upon oxidative stress treatment. Based on accurate mass measurements and MS/MS spectral matching with reference standard solutions, 10 metabolites were clearly identified. Derivatized compounds were found to have diagnostic fragment ions from the reagent itself, as well as structurally informative ions useful for metabolite identification.
  
  
  CONCLUSIONS
  This simple derivatization reaction can be applied to the relative quantitation of amine-, thiol- and phenol-containing compounds, with improved sensitivity and chromatographic peak shapes due to the increased hydrophobicity of polar metabolites not readily amenable to reversed-phase LC/MS analysis.
authors.affiliation: Université du Québec à Montréal, Chemistry Department/Pharmaqam, Montréal, QC, Canada.
authors.forename: Michel
authors.initials: M
authors.lastname: Wagner
authors.affiliation: Université du Québec à Montréal, Chemistry Department/Pharmaqam, Montréal, QC, Canada.
authors.forename: Leanne B
authors.initials: LB
authors.lastname: Ohlund
authors.affiliation: Université du Québec à Montréal, Chemistry Department/Pharmaqam, Montréal, QC, Canada.
authors.forename: Tze Chieh
authors.initials: TC
authors.lastname: Shiao
authors.affiliation: Université du Québec à Montréal, Chemistry Department/Pharmaqam, Montréal, QC, Canada.
authors.forename: Amélie
authors.initials: A
authors.lastname: Vézina
authors.affiliation: Université du Québec à Montréal, Chemistry Department/Pharmaqam, Montréal, QC, Canada.
authors.forename: Borhane
authors.initials: B
authors.lastname: Annabi
authors.affiliation: Université du Québec à Montréal, Chemistry Department/Pharmaqam, Montréal, QC, Canada.
authors.forename: René
authors.initials: R
authors.lastname: Roy
authors.affiliation: Université du Québec à Montréal, Chemistry Department/Pharmaqam, Montréal, QC, Canada.
authors.forename: Lekha
authors.initials: L
authors.lastname: Sleno
mesh_terms: D002247:Carbon Isotopes / Q000032:analysis / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods*; D018922:HL-60 Cells; D006801:Humans; D007553:Isotope Labeling; D055432:Metabolomics / Q000379:methods*; D013388:Succinimides / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D002247:Carbon Isotopes; D013388:Succinimides