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Univerzita Pardubice

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Metabolomic Analysis of Human Plasma by Bioinert LC System Coupled with Ion Mobility Mass Spectrometry

Autoři: Langová Alena | Jirásko Robert | Peterka Ondřej | Šubrtová Veronika | Holčapek Michal

Rok: 2024

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
eng	Metabolomic Analysis of Human Plasma by Bioinert LC System Coupled with Ion Mobility Mass Spectrometry	Metabolomic analysis is a rapidly developing scientific discipline that deals with the complex identification and quantification of metabolites to understand metabolic pathways in organisms. Changes in the composition and concentration of metabolites can indicate various serious diseases, such as cancer, and comprehensive metabolomic analysis can help to understand dysregulations. The metabolomic analysis is mainly based on mass spectrometry coupled with LC, where metabolites often contain problematic functional groups for the separation, which may interact with the surface of the instrument. New fully bioinert system improves sensitivity of analysis due to metal-free components and the absence of iron and steel in the system. Ion mobility mass spectrometry (IM-MS) has emerged as powerful technology for metabolomic analysis, which enables separation of various isomers and provides technical advantages compare to conventional LC/MS, such as collision cross-section measurement (CCS) and signal-to-noise ratio. The bioinert Premier UHPLC system with Premier BEH Amide column (150×2.1 mm; 1.7 µm) coupled to IM-MS was used. The new separation method was developed with a total run time of 20 min, including column equilibration. The following conditions were set: gradient elution, where mobile phase A was acetonitrile with 0.005 % acetic acid (AA) and mobile phase B 15 mM ammonium acetate with 0.005 % AA, and flow rate 0.4 mL/min. The IM-MS conditions were optimized for both positive and negative ion modes. The bioinert system significantly brings improvement in sensitivity and peak shape of phosphate metabolites compare to conventional system and coupling with IM-MS enables highly confident identification. The optimized method was used for analysis of NIST SRM 1950 human plasma, and more than 100 metabolites (without lipids) were identified based on retention time, mass shift within 3 mDa tolerance in the full-scan mode, tandem mass spectrometry, retention dependencies, an	Metabolomic; Human Plasma; Bioinert System; Ion Mobility; Mass Spectrometry