出版社：American Society for Biochemistry and Molecular Biology
摘要：This article presents a novel methodology for the analysis ofethanolamine glycerophospholipid (PE) and lysoPE molecular speciesdirectly from lipid extracts of biological samples. Throughbrief treatment of lipid extracts with fluorenylmethoxylcarbonyl(Fmoc) chloride, PE and lysoPE species were selectively derivatizedto their corresponding carbamates. The reaction solution wasinfused directly into the ion source of an electrospray ionizationmass spectrometer after appropriate dilution. The facile lossof the Fmoc moiety dramatically enhanced the analytic sensitivityand allowed the identification and quantitation of low-abundancemolecular species. A detection limitation of attomoles (amoles)per microliter for PE and lysoPE analysis was readily achievedusing this technique (at least a 100-fold improvement from ourprevious method) with a >15,000-fold dynamic range. Throughintrasource separation and multidimensional mass spectrometryarray analysis of derivatized species, marked improvements insignal-to-noise ratio, molecular species identification, andquantitation can be realized. The procedure is both simple andeffective and can be extended to analyze many other lipid classesor other cellular metabolites by adjustments in specific derivatizationconditions.
Thus, through judicious derivatization, a new dimension exploitingspecific functional reactivities in each lipid class can beused in conjunction with shotgun lipidomics to penetrate fartherinto the low-abundance regime of cellular lipidomes.Abbreviations: 2D, two-dimensional; DMAP, dimethylaminopyridine; ESI, electrospray ionization; Fmoc-Cl, fluorenylmethoxylcarbonyl chloride; m:n, acyl chain containing m carbons and n double bonds; MS, mass spectrometry; NAC, 2-(2-naphthyl)acetyl chloride; NL, neutral loss; PE, ethanolamine glycerophospholipid; PI, precursor ion; PlsEtn, plasmenylethanolamine; PtdCho, phosphatidylcholine; PtdEtn, phosphatidylethanolamine; PtdGro, phosphatidylglycerol
Supplementary key words electrospray ionization-mass spectrometry • fluorenylmethoxylcarbonyl derivatization • lipidome • lipidomics • lysophosphatidylethanolamine • multidimensional mass spectrometry • plasmalogen
