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High-throughput Lipidomic Quantitation of Biological Samples
Autoři: Holčapek Michal | Wolrab Denise | Jirásko Robert | Chocholoušková Michaela | Peterka Ondřej
Rok: 2019
Druh publikace: ostatní - přednáška nebo poster
Strana od-do: nestránkováno
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Jazyk Název Abstrakt Klíčová slova
eng High-throughput Lipidomic Quantitation of Biological Samples Lipids are found in all eukaryotic cells, where they fulfill various physiological functions, and their dysregulation may be linked to serious diseases, such as cancer, cardiovascular diseases, etc. The coupling of liquid-phase separation techniques and mass spectrometry (MS) and MS alone are dominating the field of lipidomic analysis [1]. We have developed and validated several MS based methods for the high-throughput clinical lipidomic quantitation using the coupling with ultrahigh-performance supercritical fluid chromatography (UHPSFC) [2], ultrahigh-performance liquid chromatography (UHPLC) [3], the direct infusion (shotgun approach) [2], or matrix-assisted laser desorption/ionization (MALDI) coupled to high-resolution Orbitrap mass analyzer [4]. UHPSFC/MS, UHPLC/MS, and shotgun MS are applied mainly for glycerophospholipids, sphingolipids, and glycerolipids using positive-ion electrospray ionization (ESI), while MALDI is used in the negative-ion mode to obtain complementary information on sulfatides and other anionic lipid subclasses. Several hundred lipids species are typically quantified in biological samples, but the major issue in the lipidomic quantitation is the data reliability over longer period of time, the comparability of results among different groups, and the absolute molar quantitation based on the use of the right lipid class exogenous internal standards [1, 5]. The use of validated protocols in line with FDA and EMA requirements (e.g., matrix effect, recovery rate, precision, accuracy, repeatability, and selectivity), and the use of quality control samples will be discussed as well. The optimized sample preparation protocol was applied for the analysis of clinical plasma and serum samples and the SRM 1950 plasma and compared with the literature [6, 7].

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