Publikace detail

Development of Comprehensive LC/MS Approach for Clinical Lipidomic Applications and Biomarker Discovery

Autoři: Holčapek Michal | Cífková Eva | Lísa Miroslav | Červená Blanka | Ovčačíková Magdaléna

Rok: 2014

Druh publikace: ostatní do riv

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
cze	Development of Comprehensive LC/MS Approach for Clinical Lipidomic Applications and Biomarker Discovery	Lipidomics is used in clinical studies to improve the understanding how lipids function in biological systems and for the elucidation of the mechanism of lipid-related diseases. We are developing a comprehensive lipidomic approach based on the combination of several liquid chromatography - mass spectrometry (LC/MS) based methods. Total lipid extracts are separated into individual lipid classes using hydrophilic interaction liquid chromatography (HILIC) [1] or supercritical fluid chromatography (SFC) and quantified by response factor approach for lipid classes [2]. HILIC-LC/MS method enables the separation of multiple polar lipid classes (such as different classes of phospholipids, sphingolipids and glycolipids), but unfortunately nonpolar lipid classes coelute in the void volume of the system, so another normal-phase LC/MS method has been developed for the quantitation of tri-, di- and monoacylglycerols, cholesterol esters and cholesterol. SFC/MS is enables the separation of both polar and nonpolar lipid classes in significantly shorter analysis time. Reversed-phase UHPLC/MS method with very high peak capacity is used either as independent method or in 2D-LC/MS configuration for the most detailed lipidomic characterization. Another separation dimension can be obtained by the use of ion mobility providing both class and species separation. The multivariate data analysis using orthogonal 2 projections to latent structures (O2PLS) is used for the differentiation of human breast cancer and surrounding normal tissues. Two clinical applications (breast cancer and cardiovascular diseases) of our methodology will be shown.
eng	Development of Comprehensive LC/MS Approach for Clinical Lipidomic Applications and Biomarker Discovery	Lipidomics is used in clinical studies to improve the understanding how lipids function in biological systems and for the elucidation of the mechanism of lipid-related diseases. We are developing a comprehensive lipidomic approach based on the combination of several liquid chromatography - mass spectrometry (LC/MS) based methods. Total lipid extracts are separated into individual lipid classes using hydrophilic interaction liquid chromatography (HILIC) [1] or supercritical fluid chromatography (SFC) and quantified by response factor approach for lipid classes [2]. HILIC-LC/MS method enables the separation of multiple polar lipid classes (such as different classes of phospholipids, sphingolipids and glycolipids), but unfortunately nonpolar lipid classes coelute in the void volume of the system, so another normal-phase LC/MS method has been developed for the quantitation of tri-, di- and monoacylglycerols, cholesterol esters and cholesterol. SFC/MS is enables the separation of both polar and nonpolar lipid classes in significantly shorter analysis time. Reversed-phase UHPLC/MS method with very high peak capacity is used either as independent method or in 2D-LC/MS configuration for the most detailed lipidomic characterization. Another separation dimension can be obtained by the use of ion mobility providing both class and species separation. The multivariate data analysis using orthogonal 2 projections to latent structures (O2PLS) is used for the differentiation of human breast cancer and surrounding normal tissues. Two clinical applications (breast cancer and cardiovascular diseases) of our methodology will be shown.	LC/MS; Lipidomic; Biomarker

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