Enhanced standard operating procedures for 31P NMR-based metabolomics in tissue extracts

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dc.contributor.authorMartín Ramos, Sara
dc.contributor.authorBilbao García, Jon
dc.contributor.authorDiercks, Tammo
dc.contributor.authorMato, José María
dc.contributor.authorBernardo Seisdedos, Ganeko
dc.contributor.authorMillet, Óscar
dc.date.accessioned2025-10-16T14:51:31Z
dc.date.available2025-10-16T14:51:31Z
dc.date.issued2025-04-13
dc.date.updated2025-10-16T14:51:31Z
dc.description.abstractPhosphorylated metabolites, here referred to as phosphometabolites, are sufficiently abundant and widely distributed to provide a condensed representation of metabolism that can be readily accessed through NMR spectroscopy. This study addresses the challenge of precisely quantifying phosphometabolites via quantitative 31P NMR from tissue extracts. We optimized standard operating procedures for enhanced spectral resolution, signal intensity, and accuracy. By amply evaluating solvent and buffer conditions, reference compounds, and paramagnetic relaxation enhancers, we identified optimal conditions for metabolite analysis, including the use of trimethylphosphine oxide for accurate signal referencing due to its short T1 relaxation time and minimal offset and glycine buffer (in D2O) at pD 9.5, where virtually invariant 31P signal frequencies and sample osmolarities are observed, along with maximal NMR detection sensitivity and temperature stability of the pH. These methodological advancements significantly improve the reliability and reproducibility of phosphometabolite characterization, allowing the assignment of up to 60 independent signals in the one-dimensional (1D) 31P spectrum (of a total of 94 peaks), that resulted in the proper quantification of 44 phosphometabolites from different tissular samples.en
dc.description.sponsorshipSupport was provided by the Department of Economic Development and Infrastructures of the Government of the Autonomous Community of the Basque Country (Elkartek bg2021, bg2023). O.M. and J.M.M. acknowledge CIBERehd and the Agencia Estatal de Investigación (Spain) for grants, RTI2018-101269−B-I00, PID2021-124171OB-I00, and CEX2021-001136-S and NIH for grant NIH 1R01DK119437-01A1en
dc.identifier.citationMartin-Ramos, S., Bilbao, J., Diercks, T., Mato, J. M., Bernardo-Seisdedos, G., & Millet, O. (2025). Enhanced standard operating procedures for 31P NMR-based metabolomics in tissue extracts. JACS Au, 5(5), 2285-2293. https://doi.org/10.1021/JACSAU.5C00234
dc.identifier.doi10.1021/JACSAU.5C00234
dc.identifier.eissn2691-3704
dc.identifier.urihttps://hdl.handle.net/20.500.14454/4003
dc.language.isoeng
dc.publisherAmerican Chemical Society
dc.rights© 2025 The Authors
dc.subject.other31P NMR
dc.subject.otherGadoteridol
dc.subject.otherGlycine buffer
dc.subject.otherMetabolites
dc.subject.otherPhosphorylated metabolites
dc.subject.otherSOP
dc.subject.otherTissue analysis
dc.titleEnhanced standard operating procedures for 31P NMR-based metabolomics in tissue extractsen
dc.typejournal article
dcterms.accessRightsopen access
oaire.citation.endPage2293
oaire.citation.issue5
oaire.citation.startPage2285
oaire.citation.titleJACS Au
oaire.citation.volume5
oaire.licenseConditionhttps://creativecommons.org/licenses/by-nc-nd/4.0/
oaire.versionVoR
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