Polar metabolomics using trichloroacetic acid extraction and porous graphitic carbon stationary phase.

Autor: Day F; Liggins Institute, The University of Auckland, Auckland, New Zealand., O'Sullivan J; Liggins Institute, The University of Auckland, Auckland, New Zealand.; The Maurice Wilkins Centre, The University of Auckland, Auckland, New Zealand.; MRC Lifecourse Epidemiology Unit, University of Southampton, Southampton, UK.; Australian Parkinson's Mission, Garvan Institute of Medical Research, 384 Victoria Street, Sydney, Darlinghurst, NSW, 2010, Australia.; A*STAR Singapore Institute for Clinical Sciences, Singapore, Singapore., Ramzan F; Liggins Institute, The University of Auckland, Auckland, New Zealand., Pook C; Liggins Institute, The University of Auckland, Auckland, New Zealand. chris.pook@auckland.ac.nz.; School of Chemical Sciences, University of Auckland, 23 Symonds St., Auckland, 1010, New Zealand. chris.pook@auckland.ac.nz.
Jazyk: angličtina
Zdroj: Metabolomics : Official journal of the Metabolomic Society [Metabolomics] 2024 Jul 16; Vol. 20 (4), pp. 77. Date of Electronic Publication: 2024 Jul 16.
DOI: 10.1007/s11306-024-02146-7
Abstrakt: Introduction: Accurately identifying and quantifying polar metabolites using untargeted metabolomics has proven challenging in comparison to mid to non-polar metabolites. Hydrophilic interaction chromatography and gas chromatography-mass spectrometry are predominantly used to target polar metabolites.
Objectives: This study aims to demonstrate a simple one-step extraction combined with liquid chromatography-mass spectrometry (LC-MS) that reliably retains polar metabolites.
Methods: The method involves a MilliQ + 10% trichloroacetic acid extraction from 6 healthy individuals serum, combined with porous graphitic carbon liquid chromatography-mass spectrometry (LC-MS). The coefficient of variation (CV) assessed retention reliability of polar metabolites with logP as low as - 9. QreSS (Quantification, Retention, and System Suitability) internal standards determined the method's consistency and recovery efficiency.
Results: The method demonstrated reliable retention (CV < 0.30) of polar metabolites within a logP range of - 9.1 to 5.6. QreSS internal standards confirmed consistent performance (CV < 0.16) and effective recovery (70-130%) of polar to mid-polar metabolites. Quality control dilution series demonstrated that ~ 80% of annotated metabolites could be accurately quantified (Pearson's correlation coefficient > 0.80) within their concentration range. Repeatability was demonstrated through clustering of repeated extractions from a single sample.
Conclusion: This LC-MS method is better suited to covering the polar segment of the metabolome than current methods, offering a reliable and efficient approach for accurate quantification of polar metabolites in untargeted metabolomics.
(© 2024. The Author(s).)
Databáze: MEDLINE
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