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Trapped Ion Mobility Separation as an Additional Dimension for Better Coverage of Isomeric Lipids and Metabolites in UHPLC/MS of Biological Samples

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

Rok: 2024

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

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Jazyk	Název	Abstrakt	Klíčová slova
eng	Trapped Ion Mobility Separation as an Additional Dimension for Better Coverage of Isomeric Lipids and Metabolites in UHPLC/MS of Biological Samples	Ultrahigh-performance liquid chromatography coupled with mass spectrometry (UHPLC/MS) is one of the major approaches used in lipidomics and metabolomics for qualitative and quantitative analyses. Reversed-phase liquid chromatography and hydrophilic interaction chromatography are two main separation modes in this area. However, despite the high efficiency of chromatographic separation resulting in the high number of identifications, there are still many isomers that are not properly resolved by general methods. The additional dimension of information can be obtained by ion mobility separation performed in the gas phase between the ion source and the mass analyzer of the mass spectrometer, which fits perfectly to the time scale between the chromatography and the MS detection. In this work, we used the trapped ion mobility (TIMS) separation for distinguishing various isomers of lipids and small metabolites. An important requirement is the optimization of TIMS settings (mobility ramp time, accumulation time and mobility range) to achieve high-quality ion mobility separation and to keep fast scanning, ensuring the compatibility with UHPLC/MS. The optimized method was used for the analysis of a mixture of lipid and metabolic standards, human plasma, and tissue samples, providing highly reliable identification based on retention time, cross section values from ion mobility separation, and masses of observed ions. Compared to our previous methods without TIMS [1, 2], an increase in the number of identified lipids and metabolites was achieved. The benefit of ion mobility implemented in UHPLC/MS is illustrated in an example of separation of selected lipid and metabolite isomeric species, e.g., glucosylceramide and galactosylceramide, isomeric leucines, etc.	Ion Mobility; Separation; Isomeric Lipids; Metabolites; UHPLC/MS; Biological Samples

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