A comprehensive method for lipid profiling by liquid chromatography-ion cyclotron resonance mass spectrometry

被引:114
作者
Fauland, Alexander [4 ]
Koefeler, Harald [1 ]
Troetzmueller, Martin [1 ]
Knopf, Astrid [1 ]
Hartler, Juergen [5 ]
Eberl, Anita [1 ]
Chitraju, Chandramohan [3 ]
Lankmayr, Ernst [4 ]
Spener, Friedrich [2 ,3 ]
机构
[1] Med Univ Graz, Core Facil Mass Spectrometry, Med Res Ctr, Graz, Austria
[2] Med Univ Graz, Dept Mol Biol & Biochem, Graz, Austria
[3] Graz Univ, Dept Mol Biosci, Graz, Austria
[4] Graz Univ Technol, Inst Analyt Chem & Food Chem, A-8010 Graz, Austria
[5] Graz Univ Technol, Inst Genom & Bioinformat, A-8010 Graz, Austria
关键词
high-resolution mass spectrometry; lipidomics; HPLC/ESI-MS; HILIC; lipid droplets; triacylglycerols; phospholipids; SHOTGUN LIPIDOMICS; BIOLOGICAL SAMPLES; CRUDE EXTRACTS; QUANTIFICATION; IDENTIFICATION; QUANTITATION; SEPARATION; REVEALS;
D O I
10.1194/jlr.D016550
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
070307 [化学生物学]; 071010 [生物化学与分子生物学];
摘要
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.-Fauland, A., H. Kofeler, M. Trotzmuller, A. Knopf, J. Hartler, A. Eberl, C. Chitraju, E. Lankmayr, and F. Spener. A comprehensive method for lipid profiling by liquid chromatography-ion cyclotron resonance mass spectrometry. J. Lipid Res. 2011. 52: 2314-2322.
引用
收藏
页码:2314 / 2322
页数:9
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