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HILIC-HPLC/ESI–MS analysis of acidic glycosphingolipids in human tissues and body fluids

Autoři: Hájek Roman | Jirásko Robert | Cífková Eva | Lísa Miroslav | Holčapek Michal

Rok: 2016

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

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Jazyk	Název	Abstrakt	Klíčová slova
eng	HILIC-HPLC/ESI–MS analysis of acidic glycosphingolipids in human tissues and body fluids	Acidic glycosphingolipids are a class of sphingolipids characterized by a ceramide lipid backbone attached to an acidic saccharide chain, such as lactose sulfate (sulfatides) or anionic oligosaccharide composed of at least one sialic acid (gangliosides). These lipids play major roles in signal transduction, cell adhesion, modulating growth, factor/hormone receptor, antigen recognition and other biological functions. Recently, changes in glycosphingolipid profiles have been recorded in connection with various diseases, e.g., Fabry disease, Alzheimer’s disease or cancer. Their profiling and its application in clinical studies can reveal new information and clarify the dysregulation in the lipid metabolism during the disease. Aim of our work was to develop high-throughput method for profiling sulfatides and gangliosides in extracts of human plasma, urine, erythrocytes and selected tissues using hydrophilic interaction liquid chromatography (HILIC) and electrospray ionization mass spectrometry (ESI-MS). The extraction of real tissue samples was optimized and followed by the purification of collected aqueous phase on C18 SPE column to remove undesired salts before LC-MS. The optimization ofthe organic solvent content, the buffer type, its concentration and the mobile phase pH was performed to achieve the optimal HILIC separation with regard to the best sensitivity and separation of studied compounds. Particular lipids were identified based on the accurate m/z values of deprotonated molecules in negative-ion ESI-MS mode using high mass accuracy measurements performed on microQTOF II (Bruker Daltonics, Germany).

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