Profiling of lipid species by normal-phase liquid chromatography, nanoelectrospray ionization, and ion trap-orbitrap mass spectrometry
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- 作者：Elena Sokol ; Reinaldo Almeida ; Hans Kristian Hannibal-Bach ; Dorota Kotowska ; Johannes Vogt ; Jan Baumgart ; Karsten Kristiansen ; Robert Nitsch ; Jens Knudsen ; Christer S. Ejsing
- 关键词：Lipidomics ; Normal-phase LC ; Nanoelectrospray ionization ; Orbitrap mass spectrometry
- 刊名：Analytical Biochemistry
- 出版年：2013
- 出版时间：1 December, 2013
- 年：2013
- 卷：443
- 期：1
- 页码：88-96
- 全文大小：1112 K
文摘
Detailed analysis of lipid species can be challenging due to their structural diversity and wide concentration range in cells, tissues, and biofluids. To address these analytical challenges, we devised a reproducible, sensitive, and integrated lipidomics workflow based on normal-phase liquid chromatography-Fourier transform mass spectrometry (LC-FTMS) and LC-ITMS2 (ion trap tandem mass spectrometry) for profiling and structural analysis of lipid species. The workflow uses a normal-phase LC system for efficient separation of apolar and polar lipid species combined with sensitive and specific analysis powered by a chip-based nanoelectrospray ion source and a hybrid ion trap-orbitrap mass spectrometer. The workflow was executed using a primary LC-FTMS survey routine for identification and profiling of lipid species based on high-mass accuracy and retention time followed by a targeted LC-ITMS2 routine for characterizing the fatty acid moieties of identified lipid species. We benchmarked the performance of the workflow by characterizing the chromatographic properties of the LC-MS system for general lipid analysis. In addition, we demonstrate the efficacy of the workflow by reporting a study of low-abundant triacylglycerol and ceramide species in mouse brain cerebellum and 3T3-L1 adipocytes, respectively. The workflow described here is generic and can be extended for detailed lipid analysis of sample matrices having a wide range of lipid compositions.