Targeted mass spectrometry method for the determination of multiple gut-microbiota metabolites in human plasma

Fernandes, Sara R.; Barreiros, Luísa; Azorín, Cristian; Silva, Eduarda M.P.; Segundo, Marcela A.; Barreiros, Luisa

Publicação

Targeted mass spectrometry method for the determination of multiple gut-microbiota metabolites in human plasma

2025-11-01Artigo científico

dc.contributor.author	Fernandes, Sara R.
dc.contributor.author	Barreiros, Luísa
dc.contributor.author	Azorín, Cristian
dc.contributor.author	Silva, Eduarda M.P.
dc.contributor.author	Segundo, Marcela A.
dc.contributor.author	Barreiros, Luisa
dc.date.accessioned	2026-05-29T13:35:40Z
dc.date.available	2026-05-29T13:35:40Z
dc.date.issued	2025-11-01
dc.description.abstract	The gut microbiota profoundly impacts human health by producing metabolites that can act as biomarkers for disease diagnosis and therapy. However, accurately measuring these metabolites in biomatrices is challenging due to their low concentrations, high molecular diversity, and interference from matrix components, demanding advanced and precise analytical methodologies. Hence, an ultra-high-performance liquid chromatography method coupled to triple quadrupole-tandem mass spectrometry detection, combined with a chemical derivatization procedure, was developed and validated to quantify seven gut metabolites, namely acetic acid, propionic acid, butyric acid, p-cresol sulfate, 3-indoxyl sulfate, indole-3-acetic acid, and L-tryptophan, in human plasma. Samples were prepared by protein precipitation with acetonitrile and subsequently derivatized using 3-nitrophenylhydrazine. Chromatographic separation was achieved using a BEH C18 column, with elution performed at a f low rate of 0.2mLmin 1 and in gradient mode using formic acid-water (1:1000, v/v) and formic acid- acetonitrile (1:1000, v/v) as mobile phase components. The mass spectrometer was operated in negative ionization mode and data was acquired in selected reaction monitoring. Good linearity was achieved (r 2 >0.997) for all the target gut metabolites in the evaluated concentration ranges, with low LLOQ values (0.4–8 M). The method proved to be accurate (87.0–114 %) and precise (CV ≤ 13.5 %), achieving a score of 65 in the Blue Applicability Grade Index (BAGI) metric, which confirmed its practicality. The developed method was ultimately employed to the analysis of plasma samples from children and adults involved in clinical studies, demonstrating its usefulness in medical research.	eng
dc.identifier.citation	Fernandes, S. R., Azorín, C., Silva, E. M. P., Miró, M., Barreiros, L., & Segundo, M. A. (2025). Targeted mass spectrometry method for the determination of multiple gut-microbiota metabolites in human plasma. Journal of Chromatography B, 1265, 124764. https://doi.org/10.1016/j.jchromb.2025.124764
dc.identifier.doi	10.1016/j.jchromb.2025.124764
dc.identifier.eissn	1873-376X
dc.identifier.issn	0378-4347
dc.identifier.uri	http://hdl.handle.net/10400.22/32465
dc.language.iso	eng
dc.peerreviewed	yes
dc.publisher	Elsevier
dc.relation	project UID/50006; SFRH/ BD/130948/2017 and COVID/BD/152406/2022
dc.relation.hasversion	https://www.sciencedirect.com/science/article/pii/S1570023225003186?via%3Dihub
dc.rights.uri	http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject	Gut metabolites
dc.subject	Chemical derivatization
dc.subject	Targeted analysis
dc.subject	Mass spectrometry
dc.subject	Biomatrices
dc.subject	Clinical diagnostics
dc.title	Targeted mass spectrometry method for the determination of multiple gut-microbiota metabolites in human plasma	eng
dc.type	journal article
dspace.entity.type	Publication
oaire.citation.title	Journal of Chromatography B
oaire.citation.volume	1265
oaire.version	http://purl.org/coar/version/c_970fb48d4fbd8a85
person.familyName	Barreiros
person.givenName	Luisa
person.identifier.ciencia-id	611F-E0C5-0230
person.identifier.orcid	0000-0003-3481-5809
person.identifier.rid	D-7950-2013
person.identifier.scopus-author-id	6508205485
relation.isAuthorOfPublication	1e66bacc-64de-4ecb-96b7-4c0e366cba57
relation.isAuthorOfPublication.latestForDiscovery	1e66bacc-64de-4ecb-96b7-4c0e366cba57