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Lipidomics: Ultrahigh-Performance Supercritical Fluid Chromatography –Mass Spectrometry as a New Tool for High-Throughput and Comprehensive Analysis of Lipids
Authors: Lísa Miroslav | Holčapek Michal
Year: 2016
Type of publication: ostatní - přednáška nebo poster
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
eng Lipidomics: Ultrahigh-Performance Supercritical Fluid Chromatography –Mass Spectrometry as a New Tool for High-Throughput and Comprehensive Analysis of Lipids Lipids are biologically important compounds. Their metabolism plays a crucial role in the development of some serious human diseases, such as cardiovascular diseases, diabetes, cancer, etc. In current lipidomics, precise, comprehensive and high-throughput methods are required for the analysis of large sample sets in the clinical research, which is challenging for analytical techniques due to the enormous complexity of lipid structures. Two well-established approaches are commonly used in lipidomics, such as shotgun and LC/MS, both having some advantages and limitations. We have developed a new high-throughput and comprehensive approach for analysis of lipids in biological samples using ultrahigh-performance supercritical fluid chromatography (UHPSFC) followed by ESI-MS identification and quantitation. Developed UHPSFC method is based on 1.7 μm particle bridged ethylene hybrid silica column using the gradient of methanol – water – ammonium acetate mixture as a modifier providing separation of total lipid extract into lipid classes. Individual parameters of UHPSFC analysis have been carefully optimized to achieve a maximum number of separated lipid classes. Final UHPSFC method enables a fast separation up to 28 nonpolar and polar lipid classes within 6 minute analysis including the partial separation of species inside individual classes. Mass spectra with high mass accuracy and high resolving power are acquired using ESI in both positive- and negative-ion modes for the identification of individual species. The quantitative analysis is performed using internal standards for each lipid class added during the extraction step. Developed UHPSFC/MS method is validated and applied for the comprehensive analysis of lipid composition in real biological samples, such as plasma or normal and cancer tissues

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