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Comparison of Bioinert and Conventional UHPLC Systems for Metabolomic Analysis

Authors: Langová Alena | Peterka Ondřej | Jirásko Robert | Holčapek Michal

Year: 2024

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	Comparison of Bioinert and Conventional UHPLC Systems for Metabolomic Analysis	In recent years, metabolomics has been ranked as one of the leading omics approaches, offering direct information about the current state of the organism. This approach is utilized to identify and quantify metabolites in biological samples. Metabolomics might improve the understanding of physiological conditions and facilitate the characterization of metabolic alterations linked to serious diseases or the exploration of novel biomarkers. Metabolites are small molecules mostly containing polar function groups, e.g., phosphate and sulfate. Those ionic groups are typically challenging for chromatography because of the interaction with the metal surface of the instrument, resulting in poor peak shapes, reduced chromatographic resolution, and decreased sensitivity. A fully bioinert system compared to a conventional system, can solve these problems, especially for nucleotides containing one to three phosphate groups (e.g., AMP, ADP, ATP). Traditionally, bioinert columns are made from polyether ether ketone (PEEK), but materials like titanium and MP35N alloys are frequently favored over PEEK due to their superior capacity to resist high pressures. In this study, a new metabolomics method was developed that focuses on ionic metabolites present in human plasma. This method can separate all proteinogenic amino acids, including isomers, such as leucine/isoleucine/norleucine, carnitines, nucleotides, and nucleosides. Optimized method was applied to NIST SRM 1950 human plasma, leading to the identification of more than 130 metabolites (in addition to lipids). The identification was based on retention times, mass shifts within a 3 mDa tolerance in full-scan mode, tandem mass spectrometry, and retention dependencies. At the same time, we cross-compared two different systems: bioinert and conventional, which included both bioinert and conventional columns. The results indicate that a fully bioinert system significantly improves peak shapes and resolution. In comparison to the effe	metabolomics; UHPLC