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Optimization of UHPSFC/ESI-MS and application of Method for Characterization of Biological Samples
Authors: Peterka Ondřej | Wolrab Denise | Kuchař Ladislav | Holčapek Michal
Year: 2018
Type of publication: ostatní - přednáška nebo poster
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
eng Optimization of UHPSFC/ESI-MS and application of Method for Characterization of Biological Samples Supercritical fluid chromatography (SFC) is preferably used for analysis of non-polar substance, because the main component of mobile phase is supercritical carbon dioxide. The addition of organic modifiers enhances also the determination of polar compounds. The connection of mass spectrometry (MS) and ultrahigh-performance supercritical fluid chromatography (UHPSFC) with sub-2 µm particles as stationary phase has higher separation efficiency and faster analysis times, which can be applied to lipidomics analysis. Nowadays, the instrumentation includes splitter before the backpressure regulator, which enables employing a make-up solvent for a higher ionization efficiency. Goals were optimization of parameters affecting the sensitivity of nonpolar and polar lipid in short analysis times. Pooled sample was used for the method development and optimization. Lipids were quantified using mixture of internal standards (IS). The increasing of sensitivity by makeup solvents were verified by employing various additives (acetic acid, formic acid, ammonium formate, and ammonium acetate) and flow rates (0.1, 0.25, and 0.5 mL/min). Injection volume and acquisition mode were also investigated. Data were processed by Water´s tools and our laboratory-made software LipidQuant was used for the semi-automated identification, isotopic correction, and quantitation of lipid species. Optimized method was applied to biological samples, and the results were statistically analyzed. UHPSFC/ESI-MS; Biological Samples

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