Author, as appears in the article.: Capellades J; Junza A; Samino S; Brunner JS; Schabbauer G; Vinaixa M; Yanes O
Department: Enginyeria Electrònica, Elèctrica i Automàtica
e-ISSN: 1520-6882
URV's Author/s: De la Varga Pastor, Aitana / Junza Martínez, Alexandra / SAMINO GENÉ, SARA / Vinaixa Crevillent, Maria / Yanes Torrado, Óscar
Abstract: © Isotopic-labeling experiments have been valuable to monitor the flux of metabolic reactions in biological systems, which is crucial to understand homeostatic alterations with disease. Experimental determination of metabolic fluxes can be inferred from a characteristic rearrangement of stable isotope tracers (e.g., 13C or 15N) that can be detected by mass spectrometry (MS). Metabolites measured are generally members of well-known metabolic pathways, and most of them can be detected using both gas chromatography (GC)-MS and liquid chromatography (LC)-MS. In here, we show that GC methods coupled to chemical ionization (CI) MS have a clear advantage over alternative methodologies due to GC's superior chromatography separation efficiency and the fact that CI is a soft ionization technique that yields identifiable protonated molecular ion peaks. We tested diverse GC-CI-MS setups, including methane and isobutane reagent gases, triple quadrupole (QqQ) MS in SIM mode, or selected ion clusters using optimized narrow windows (∼10 Da) in scan mode, and standard full scan methods using high resolution GC-(q)TOF and GC-Orbitrap systems. Isobutane as a reagent gas in combination with both low-resolution (LR) and high-resolution (HR) MS showed the best performance, enabling precise detection of isotopologues in most metabolic intermediates of central carbon metabolism. Finally, with the aim of overcoming manual operations, we developed an R-based tool called isoSCAN that automatically quantifies all isotopologues of intermediate metabolites of glycolysis, TCA cycle, amino acids, pentose phosphate pathway, and urea cycle, from LRMS and HRMS data.
Thematic Areas: Química Medicina ii Medicina i Materiais Interdisciplinar Geociências General medicine Farmacia Ensino Engenharias iv Engenharias iii Engenharias ii Enfermagem Ciências biológicas iii Ciências biológicas ii Ciências biológicas i Ciências ambientais Ciências agrárias i Ciência de alimentos Ciência da computação Chemistry, analytical Biotecnología Biodiversidade Astronomia / física Analytical chemistry
licence for use: https://creativecommons.org/licenses/by/3.0/es/
ISSN: 0003-2700
Author's mail: maria.vinaixa@urv.cat oscar.yanes@urv.cat alexandra.junza@urv.cat aitana.delavarga@urv.cat maria.vinaixa@urv.cat
Author identifier: 0000-0001-9804-0171 0000-0003-3695-7157 0000-0001-7205-0419 0000-0002-3124-4409 0000-0001-9804-0171
Record's date: 2023-12-16
Papper version: info:eu-repo/semantics/acceptedVersion
Link to the original source: https://pubs.acs.org/doi/10.1021/acs.analchem.0c02998
Licence document URL: http://repositori.urv.cat/ca/proteccio-de-dades/
Papper original source: Analytical Chemistry. 93 (3): 1242-1248
APA: Capellades J; Junza A; Samino S; Brunner JS; Schabbauer G; Vinaixa M; Yanes O (2021). Exploring the Use of Gas Chromatography Coupled to Chemical Ionization Mass Spectrometry (GC-CI-MS) for Stable Isotope Labeling in Metabolomics. Analytical Chemistry, 93(3), 1242-1248. DOI: 10.1021/acs.analchem.0c02998
Article's DOI: 10.1021/acs.analchem.0c02998
Entity: Universitat Rovira i Virgili
Journal publication year: 2021
Publication Type: Journal Publications
Repositori URV