title: A comprehensive method for lipid profiling by liquid chromatography-ion cyclotron resonance mass spectrometry.
abstract: This work aims to combine chromatographic retention, high mass resolution and accuracy, MS/MS spectra, and a package for automated identification and quantitation of lipid species in one platform for lipidomic analysis. The instrumental setup elaborated comprises reversed-phase HPLC coupled to a Fourier transform ion cyclotron resonance mass spectrometer (LTQ-FT), and Lipid Data Analyzer (LDA) software. Data analysis for lipid species quantification in this platform is based on retention time, mass resolution of 200,000, and mass accuracy below 2 ppm. In addition, automatically generated MS/MS spectra provide structural information at molecular level. This LC/MS technology allows analyzing complex biological samples in a quantitative manner as shown here paradigmatically for murine lipid droplets having a huge surplus of triacylglycerol species. Chromatographic preseparation of the bulk lipid class alleviates the problem of ion suppression of lipid species from other classes. Extension of 1D to 2D chromatography is possible, yet time consuming. The platform affords unambiguous detection of lipid species as low as 0.1‰ within major lipid classes. Taken together, a novel lipidomic LC/MS platform based on chromatographic retention, high mass resolution and accuracy, MS/MS analysis, and quantitation software enables analysis of complex samples as demonstrated for lipid droplets.
authors.affiliation: Institute of Analytical Chemistry and Food Chemistry Graz University of Technology, Graz, Austria.
authors.forename: Alexander
authors.initials: A
authors.lastname: Fauland
authors.affiliation: Core Facility for Mass Spectrometry, Center for Medical Research, Medical University of Graz, Graz, Austria. Electronic address: harald.koefeler@medunigraz.at.
authors.forename: Harald
authors.initials: H
authors.lastname: Köfeler
authors.affiliation: Core Facility for Mass Spectrometry, Center for Medical Research, Medical University of Graz, Graz, Austria.
authors.forename: Martin
authors.initials: M
authors.lastname: Trötzmüller
authors.affiliation: Core Facility for Mass Spectrometry, Center for Medical Research, Medical University of Graz, Graz, Austria.
authors.forename: Astrid
authors.initials: A
authors.lastname: Knopf
authors.affiliation: Institute for Genomics and Bioinformatics, Graz University of Technology, Graz, Austria.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hartler
authors.affiliation: Core Facility for Mass Spectrometry, Center for Medical Research, Medical University of Graz, Graz, Austria.
authors.forename: Anita
authors.initials: A
authors.lastname: Eberl
authors.affiliation: Department of Molecular Biosciences University of Graz, Graz, Austria.
authors.forename: Chandramohan
authors.initials: C
authors.lastname: Chitraju
authors.affiliation: Institute of Analytical Chemistry and Food Chemistry Graz University of Technology, Graz, Austria.
authors.forename: Ernst
authors.initials: E
authors.lastname: Lankmayr
authors.affiliation: Department of Molecular Biology and Biochemistry Medical University of Graz, Graz, Austria; Department of Molecular Biosciences University of Graz, Graz, Austria.
authors.forename: Friedrich
authors.initials: F
authors.lastname: Spener
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase; D017357:Cyclotrons; D008055:Lipids / Q000032:analysis* / Q000737:chemistry*; D051379:Mice; D008810:Mice, Inbred C57BL; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D013997:Time Factors
chemical_list: D008055:Lipids