A comparison of hydrophilic interaction liquid chromatography and capillary electrophoresis for the metabolomics analysis of human serum.

  • Acknowledgements: Project PID2020–120020RA-I00 funded by MCIN/AEI/ 10.13039/501100011033 ‘A way to make Europe’ and Project PROYEXCEL_00195 (Andalusia Government, Spain). LN thanks a grant for the Grants for the requalification of the Spanish University System for 2021–2023 (Maria Zambrano). MMDP gratefully acknowledges the European Social Fund and the Junta de Andalucía for financing her postdoctoral contract (DOC_00230). MHM gratefully acknowledges the grant IJC2019–040989-I funded by MCIN/AEI/ 10.13039/501100011033.
  • Authors: L. Narduzzi, M.M. Delgado-Povedano, F.J. Lara, B. Le Bizec, A.M. García-Campaña, G. Dervilly, M. Hernández-Mesa.
  • Reference: Journal of Chromatography A 1706 (2023) 464239.

Cationic, anionic, zwitterionic and, partially polar metabolites are very important constituents of blood serum. Several of these metabolites underpin the core metabolism of cells (e.g., Krebs cycle, urea cycle, proteins synthesis, etc.), while others might be considered ancillary but still important to grasp the status of any organism through blood serum analysis. Due to its wide chemical diversity, modern metabolomics analysis of serum is still struggling to provide a complete and comprehensive picture of the polar metabolome, due to the limitations of each specific analytical method. In this study, two metabolomics-based analytical methods using the most successful techniques for polar compounds separation in human serum samples, namely hydrophilic interaction liquid chromatography (HILIC) and capillary electrophoresis (CE), are evaluated, both coupled to a high-resolution time-of-flight mass spectrometer via electrospray ionization (ESI-Q-TOF-MS). The performance of the two methods have been compared using five terms of comparison, three of which are specific to metabolomics, such as (1) compounds’ detectability (2) Pezzatti score (Pezzatti et al. 2018), (3) intra-day precision (repeatability), (4) ease of automatic analysis of the data (through a common deconvolution alignment and extrapolation software, MS-DIAL, and (5) time & cost analysis. From this study, HILIC-MS proved to be a better tool for polar metabolome analysis, while CE-MS helped identify some interesting variables that gave it interest in completing metabolome coverage in metabolomics studies. Finally, in this framework, MS-DIAL demonstrates for the first time its ability to process CE data for metabolomics, although it is not designed for it.

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