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Ultrahigh performance supercritical fluid chromatography – mass spectrometry: new approach for high throughput and comprehensive lipidomic analysis

Autoři: Holčapek Michal | Lísa Miroslav | Khalikova Maria

Rok: 2016

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

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Jazyk	Název	Abstrakt	Klíčová slova
eng	Ultrahigh performance supercritical fluid chromatography – mass spectrometry: new approach for high throughput and comprehensive lipidomic analysis	Total lipid extracts from selected human tissues and bo dy fluids (plasma and urine) are prepared by modified Folch procedure using chloroform – methanol – water extraction. UHPSFC experiments were performed on UPC 2 instrument (Waters) using 1.7 μ m particle bridged ethylene hybrid silica column, separation temperature 60°C and gradient of methanol – water – ammonium acetate mixture as a modifier coupled with Syna pt HDMS G2Si instrument (Waters) using electrospray ionization. Indiv idual parameters of UHPSFC analysis are 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 min analysis including the partial separation of spe cies inside individual classes. Mass spectra with high mass accuracy and high resolving powe r are acquired using ESI in both positive- and negative-ion modes for the identifica tion 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 validate d and applied for the comprehensive analysis of lipid composition in real biologi cal samples, such as human plasma , normal and cancer tissues.

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