Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.22/22078 |
Resumo: | Protein-bound uremic toxins, mainly indoxyl sulfate (3-INDS), p-cresol sulfate (pCS), and indole-3-acetic acid (3-IAA) but also phenol (Pol) and p-cresol (pC), are progressively accumulated during chronic kidney disease (CKD). Their accurate measurement in biomatrices is demanded for timely diagnosis and adoption of appropriate therapeutic measures. Multiana lyte methods allowing the establishment of a uremic metabolite profle are still missing. Hence, the aim of this work was to develop a rapid and sensitive method based on high-performance liquid chromatography with fuorescence detection for the simultaneous quantifcation of Pol, 3-IAA, pC, 3-INDS, and pCS in human plasma. Separation was attained in 12 min, using a monolithic C18 column and isocratic elution with acetonitrile and phosphate bufer containing an ion-pairing reagent, at a fow rate of 2 mL min−1. Standards were prepared in plasma and quantifcation was performed using the background sub traction approach. LOQ values were≤0.2 µg mL−1 for all analytes except for pCS (LOQ of 2 µg mL−1). The method proved to be accurate (93.5–112%) and precise (CV≤14.3%). The multianalyte application of the method, associated to a reduced sample volume (50 µL), a less toxic internal standard (eugenol) in comparison to the previously applied 2,6-dimethylphenol and 4-ethylphenol, and a green extraction solvent (ethanol), resulted in the AGREE score of 0.62 which is in line with the recent trend of green and sustainable analytical chemistry. The validated method was successfully applied to the analysis of plasma samples from control subjects exhibiting normal levels of uremic toxins and CKD patients presenting signifcantly higher levels of 3-IAA, pC, 3-INDS, and pCS that can be further investigated as biomarkers of disease progression. |
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Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samplesUremic toxinIndole-3-acetic acidp-Cresolp-Cresol sulfate3-Indoxyl sulfateChronic kidney diseaseProtein-bound uremic toxins, mainly indoxyl sulfate (3-INDS), p-cresol sulfate (pCS), and indole-3-acetic acid (3-IAA) but also phenol (Pol) and p-cresol (pC), are progressively accumulated during chronic kidney disease (CKD). Their accurate measurement in biomatrices is demanded for timely diagnosis and adoption of appropriate therapeutic measures. Multiana lyte methods allowing the establishment of a uremic metabolite profle are still missing. Hence, the aim of this work was to develop a rapid and sensitive method based on high-performance liquid chromatography with fuorescence detection for the simultaneous quantifcation of Pol, 3-IAA, pC, 3-INDS, and pCS in human plasma. Separation was attained in 12 min, using a monolithic C18 column and isocratic elution with acetonitrile and phosphate bufer containing an ion-pairing reagent, at a fow rate of 2 mL min−1. Standards were prepared in plasma and quantifcation was performed using the background sub traction approach. LOQ values were≤0.2 µg mL−1 for all analytes except for pCS (LOQ of 2 µg mL−1). The method proved to be accurate (93.5–112%) and precise (CV≤14.3%). The multianalyte application of the method, associated to a reduced sample volume (50 µL), a less toxic internal standard (eugenol) in comparison to the previously applied 2,6-dimethylphenol and 4-ethylphenol, and a green extraction solvent (ethanol), resulted in the AGREE score of 0.62 which is in line with the recent trend of green and sustainable analytical chemistry. The validated method was successfully applied to the analysis of plasma samples from control subjects exhibiting normal levels of uremic toxins and CKD patients presenting signifcantly higher levels of 3-IAA, pC, 3-INDS, and pCS that can be further investigated as biomarkers of disease progression.SpringerRepositório Científico do Instituto Politécnico do PortoSilva, Luís A. P. Campagnolo, Stefano Fernandes, Sara R. Marques, Sara S. Barreiros, LuisaSampaio‑Maia, Benedita Segundo, Marcela A. 2023-02-01T15:11:21Z20232023-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.22/22078engSilva, L.A.P., Campagnolo, S., Fernandes, S.R. et al. Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples. Anal Bioanal Chem 415, 683–694 (2023). https://doi.org/10.1007/s00216-022-04458-w1618-264210.1007/s00216-022-04458-w1618-2650metadata only accessinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-03-13T13:18:27Zoai:recipp.ipp.pt:10400.22/22078Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T17:42:09.821686Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples |
title |
Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples |
spellingShingle |
Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples Silva, Luís A. P. Uremic toxin Indole-3-acetic acid p-Cresol p-Cresol sulfate 3-Indoxyl sulfate Chronic kidney disease |
title_short |
Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples |
title_full |
Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples |
title_fullStr |
Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples |
title_full_unstemmed |
Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples |
title_sort |
Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples |
author |
Silva, Luís A. P. |
author_facet |
Silva, Luís A. P. Campagnolo, Stefano Fernandes, Sara R. Marques, Sara S. Barreiros, Luisa Sampaio‑Maia, Benedita Segundo, Marcela A. |
author_role |
author |
author2 |
Campagnolo, Stefano Fernandes, Sara R. Marques, Sara S. Barreiros, Luisa Sampaio‑Maia, Benedita Segundo, Marcela A. |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Repositório Científico do Instituto Politécnico do Porto |
dc.contributor.author.fl_str_mv |
Silva, Luís A. P. Campagnolo, Stefano Fernandes, Sara R. Marques, Sara S. Barreiros, Luisa Sampaio‑Maia, Benedita Segundo, Marcela A. |
dc.subject.por.fl_str_mv |
Uremic toxin Indole-3-acetic acid p-Cresol p-Cresol sulfate 3-Indoxyl sulfate Chronic kidney disease |
topic |
Uremic toxin Indole-3-acetic acid p-Cresol p-Cresol sulfate 3-Indoxyl sulfate Chronic kidney disease |
description |
Protein-bound uremic toxins, mainly indoxyl sulfate (3-INDS), p-cresol sulfate (pCS), and indole-3-acetic acid (3-IAA) but also phenol (Pol) and p-cresol (pC), are progressively accumulated during chronic kidney disease (CKD). Their accurate measurement in biomatrices is demanded for timely diagnosis and adoption of appropriate therapeutic measures. Multiana lyte methods allowing the establishment of a uremic metabolite profle are still missing. Hence, the aim of this work was to develop a rapid and sensitive method based on high-performance liquid chromatography with fuorescence detection for the simultaneous quantifcation of Pol, 3-IAA, pC, 3-INDS, and pCS in human plasma. Separation was attained in 12 min, using a monolithic C18 column and isocratic elution with acetonitrile and phosphate bufer containing an ion-pairing reagent, at a fow rate of 2 mL min−1. Standards were prepared in plasma and quantifcation was performed using the background sub traction approach. LOQ values were≤0.2 µg mL−1 for all analytes except for pCS (LOQ of 2 µg mL−1). The method proved to be accurate (93.5–112%) and precise (CV≤14.3%). The multianalyte application of the method, associated to a reduced sample volume (50 µL), a less toxic internal standard (eugenol) in comparison to the previously applied 2,6-dimethylphenol and 4-ethylphenol, and a green extraction solvent (ethanol), resulted in the AGREE score of 0.62 which is in line with the recent trend of green and sustainable analytical chemistry. The validated method was successfully applied to the analysis of plasma samples from control subjects exhibiting normal levels of uremic toxins and CKD patients presenting signifcantly higher levels of 3-IAA, pC, 3-INDS, and pCS that can be further investigated as biomarkers of disease progression. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-02-01T15:11:21Z 2023 2023-01-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.22/22078 |
url |
http://hdl.handle.net/10400.22/22078 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Silva, L.A.P., Campagnolo, S., Fernandes, S.R. et al. Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples. Anal Bioanal Chem 415, 683–694 (2023). https://doi.org/10.1007/s00216-022-04458-w 1618-2642 10.1007/s00216-022-04458-w 1618-2650 |
dc.rights.driver.fl_str_mv |
metadata only access info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
metadata only access |
eu_rights_str_mv |
openAccess |
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 |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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