Evaluation of human plasma sample preparation protocols for untargeted metabolic profiles analyzed by UHPLC-ESI-TOF-MS

Eight human plasma preparation protocols were evaluated for their suitability for metabolomic studies by ultra-high-performance liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry: organic solvent protein precipitation (PPT) with either methanol or acetonitril...

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Detalhes bibliográficos
Autores: Rico Parrilla, Estitxu, González Mendia, Oscar, Blanco, María Encarnación, Alonso Rojas, Rosa María
Tipo de documento: artigo
Data de publicação:2014
País:España
Recursos:Universidad del País Vasco
Repositório:Addi. Archivo Digital para la Docencia y la Investigación
OAI Identifier:oai:addi.ehu.eus:10810/65673
Acesso em linha:http://hdl.handle.net/10810/65673
Access Level:Acceso aberto
Palavra-chave:metabolomics
plasma
sample treatment
reproducibility
liquid chromatography-mass spectrometry
hybridSPE
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spelling Evaluation of human plasma sample preparation protocols for untargeted metabolic profiles analyzed by UHPLC-ESI-TOF-MSRico Parrilla, EstitxuGonzález Mendia, OscarBlanco, María EncarnaciónAlonso Rojas, Rosa Maríametabolomicsplasmasample treatmentreproducibilityliquid chromatography-mass spectrometryhybridSPEEight human plasma preparation protocols were evaluated for their suitability for metabolomic studies by ultra-high-performance liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry: organic solvent protein precipitation (PPT) with either methanol or acetonitrile in 2:1 and 3:1 (v/v) ratios with plasma; solid-phase extraction (SPE) using C18 or HybridSPE cartridges; and a combination of PPT and SPE C18 cartridges and microextraction by packed sorbent. A study design in which the order of injection of the samples was not randomized is presented. The analyses were conducted in a BEH C18 column (1.7 μm, 2.1 mm × 100 mm) using a linear gradient from 100 % water to 100 % methanol, both with 0.1 % formic acid, in 21 min. The most reproducible protocol considering both the univariate and the multivariate analysis results was PPT with acetonitrile in a 2:1 (v/v) ratio with plasma, offering a mean coefficient of variation of the area of all the detected features of 0.15 and one of the best clusterings in the principal component analysis plots. On the other hand, the highest number of extracted features was achieved using methanol in a 2:1 (v/v) ratio with plasma as the PPT solvent, closely followed by the same protocol with acetonitrile in a 2:1 (v/v) ratio with plasma, which offered only 1.2 % fewer repeatable features. In terms of concentration of remaining protein, protocols based on PPT with acetonitrile provided cleaner extracts than protocols based on PPT with methanol. Finally, pairwise comparison showed that the use of PPT- and SPE-based protocols offers a different coverage of the metabolome.Springer202420242014info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10810/65673reponame:Addi. Archivo Digital para la Docencia y la Investigacióninstname:Universidad del País VascoIngléshttps://link.springer.com/article/10.1007/s00216-014-8212-yinfo:eu-repo/semantics/openAccessCopyright © 2014, Springer-Verlag Berlin Heidelbergoai:addi.ehu.eus:10810/656732026-06-18T09:23:17Z
dc.title.none.fl_str_mv Evaluation of human plasma sample preparation protocols for untargeted metabolic profiles analyzed by UHPLC-ESI-TOF-MS
title Evaluation of human plasma sample preparation protocols for untargeted metabolic profiles analyzed by UHPLC-ESI-TOF-MS
spellingShingle Evaluation of human plasma sample preparation protocols for untargeted metabolic profiles analyzed by UHPLC-ESI-TOF-MS
Rico Parrilla, Estitxu
metabolomics
plasma
sample treatment
reproducibility
liquid chromatography-mass spectrometry
hybridSPE
title_short Evaluation of human plasma sample preparation protocols for untargeted metabolic profiles analyzed by UHPLC-ESI-TOF-MS
title_full Evaluation of human plasma sample preparation protocols for untargeted metabolic profiles analyzed by UHPLC-ESI-TOF-MS
title_fullStr Evaluation of human plasma sample preparation protocols for untargeted metabolic profiles analyzed by UHPLC-ESI-TOF-MS
title_full_unstemmed Evaluation of human plasma sample preparation protocols for untargeted metabolic profiles analyzed by UHPLC-ESI-TOF-MS
title_sort Evaluation of human plasma sample preparation protocols for untargeted metabolic profiles analyzed by UHPLC-ESI-TOF-MS
dc.creator.none.fl_str_mv Rico Parrilla, Estitxu
González Mendia, Oscar
Blanco, María Encarnación
Alonso Rojas, Rosa María
author Rico Parrilla, Estitxu
author_facet Rico Parrilla, Estitxu
González Mendia, Oscar
Blanco, María Encarnación
Alonso Rojas, Rosa María
author_role author
author2 González Mendia, Oscar
Blanco, María Encarnación
Alonso Rojas, Rosa María
author2_role author
author
author
dc.subject.none.fl_str_mv metabolomics
plasma
sample treatment
reproducibility
liquid chromatography-mass spectrometry
hybridSPE
topic metabolomics
plasma
sample treatment
reproducibility
liquid chromatography-mass spectrometry
hybridSPE
description Eight human plasma preparation protocols were evaluated for their suitability for metabolomic studies by ultra-high-performance liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry: organic solvent protein precipitation (PPT) with either methanol or acetonitrile in 2:1 and 3:1 (v/v) ratios with plasma; solid-phase extraction (SPE) using C18 or HybridSPE cartridges; and a combination of PPT and SPE C18 cartridges and microextraction by packed sorbent. A study design in which the order of injection of the samples was not randomized is presented. The analyses were conducted in a BEH C18 column (1.7 μm, 2.1 mm × 100 mm) using a linear gradient from 100 % water to 100 % methanol, both with 0.1 % formic acid, in 21 min. The most reproducible protocol considering both the univariate and the multivariate analysis results was PPT with acetonitrile in a 2:1 (v/v) ratio with plasma, offering a mean coefficient of variation of the area of all the detected features of 0.15 and one of the best clusterings in the principal component analysis plots. On the other hand, the highest number of extracted features was achieved using methanol in a 2:1 (v/v) ratio with plasma as the PPT solvent, closely followed by the same protocol with acetonitrile in a 2:1 (v/v) ratio with plasma, which offered only 1.2 % fewer repeatable features. In terms of concentration of remaining protein, protocols based on PPT with acetonitrile provided cleaner extracts than protocols based on PPT with methanol. Finally, pairwise comparison showed that the use of PPT- and SPE-based protocols offers a different coverage of the metabolome.
publishDate 2014
dc.date.none.fl_str_mv 2014
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10810/65673
url http://hdl.handle.net/10810/65673
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv https://link.springer.com/article/10.1007/s00216-014-8212-y
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
Copyright © 2014, Springer-Verlag Berlin Heidelberg
eu_rights_str_mv openAccess
rights_invalid_str_mv Copyright © 2014, Springer-Verlag Berlin Heidelberg
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv reponame:Addi. Archivo Digital para la Docencia y la Investigación
instname:Universidad del País Vasco
instname_str Universidad del País Vasco
reponame_str Addi. Archivo Digital para la Docencia y la Investigación
collection Addi. Archivo Digital para la Docencia y la Investigación
repository.name.fl_str_mv
repository.mail.fl_str_mv
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