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Direct Derivatization in Dried Blood Spots for Oxidized and Reduced Glutathione Quantification in Newborns

The glutathione (GSH)-to-glutathione disulfide (GSSG) ratio is an essential node contributing to intracellular redox status. GSH/GSSG determination in whole blood can be accomplished by liquid chromatography–mass spectrometry (LC-MS) after the derivatization of GSH with N-ethylmaleimide (NEM). While...

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Autores principales: Ten-Doménech, Isabel, Solaz-García, Álvaro, Lara-Cantón, Inmaculada, Pinilla-Gonzalez, Alejandro, Parra-Llorca, Anna, Vento, Máximo, Quintás, Guillermo, Kuligowski, Julia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9219658/
https://www.ncbi.nlm.nih.gov/pubmed/35740062
http://dx.doi.org/10.3390/antiox11061165
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author Ten-Doménech, Isabel
Solaz-García, Álvaro
Lara-Cantón, Inmaculada
Pinilla-Gonzalez, Alejandro
Parra-Llorca, Anna
Vento, Máximo
Quintás, Guillermo
Kuligowski, Julia
author_facet Ten-Doménech, Isabel
Solaz-García, Álvaro
Lara-Cantón, Inmaculada
Pinilla-Gonzalez, Alejandro
Parra-Llorca, Anna
Vento, Máximo
Quintás, Guillermo
Kuligowski, Julia
author_sort Ten-Doménech, Isabel
collection PubMed
description The glutathione (GSH)-to-glutathione disulfide (GSSG) ratio is an essential node contributing to intracellular redox status. GSH/GSSG determination in whole blood can be accomplished by liquid chromatography–mass spectrometry (LC-MS) after the derivatization of GSH with N-ethylmaleimide (NEM). While this is feasible in a laboratory environment, its application in the clinical scenario is cumbersome and therefore ranges reported in similar populations differ noticeably. In this work, an LC-MS procedure for the determination of GSH and GSSG in dried blood spot (DBS) samples based on direct in situ GSH derivatization with NEM of only 10 µL of blood was developed. This novel method was applied to 73 cord blood samples and 88 residual blood volumes from routine newborn screening performed at discharge from healthy term infants. Two clinical scenarios simulating conditions of sampling and storage relevant for routine clinical analysis and clinical trials were assessed. Levels of GSH-NEM and GSSG measured in DBS samples were comparable to those obtained by liquid blood samples. GSH-NEM and GSSG median values for cord blood samples were significantly lower than those for samples at discharge. However, the GSH-NEM-to-GSSG ratios were not statistically different between both groups. With DBS testing, the immediate manipulation of samples by clinical staff is reduced. We therefore expect that this method will pave the way in providing an accurate and more robust determination of the GSH/GSSG values and trends reported in clinical trials.
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spelling pubmed-92196582022-06-24 Direct Derivatization in Dried Blood Spots for Oxidized and Reduced Glutathione Quantification in Newborns Ten-Doménech, Isabel Solaz-García, Álvaro Lara-Cantón, Inmaculada Pinilla-Gonzalez, Alejandro Parra-Llorca, Anna Vento, Máximo Quintás, Guillermo Kuligowski, Julia Antioxidants (Basel) Article The glutathione (GSH)-to-glutathione disulfide (GSSG) ratio is an essential node contributing to intracellular redox status. GSH/GSSG determination in whole blood can be accomplished by liquid chromatography–mass spectrometry (LC-MS) after the derivatization of GSH with N-ethylmaleimide (NEM). While this is feasible in a laboratory environment, its application in the clinical scenario is cumbersome and therefore ranges reported in similar populations differ noticeably. In this work, an LC-MS procedure for the determination of GSH and GSSG in dried blood spot (DBS) samples based on direct in situ GSH derivatization with NEM of only 10 µL of blood was developed. This novel method was applied to 73 cord blood samples and 88 residual blood volumes from routine newborn screening performed at discharge from healthy term infants. Two clinical scenarios simulating conditions of sampling and storage relevant for routine clinical analysis and clinical trials were assessed. Levels of GSH-NEM and GSSG measured in DBS samples were comparable to those obtained by liquid blood samples. GSH-NEM and GSSG median values for cord blood samples were significantly lower than those for samples at discharge. However, the GSH-NEM-to-GSSG ratios were not statistically different between both groups. With DBS testing, the immediate manipulation of samples by clinical staff is reduced. We therefore expect that this method will pave the way in providing an accurate and more robust determination of the GSH/GSSG values and trends reported in clinical trials. MDPI 2022-06-14 /pmc/articles/PMC9219658/ /pubmed/35740062 http://dx.doi.org/10.3390/antiox11061165 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ten-Doménech, Isabel
Solaz-García, Álvaro
Lara-Cantón, Inmaculada
Pinilla-Gonzalez, Alejandro
Parra-Llorca, Anna
Vento, Máximo
Quintás, Guillermo
Kuligowski, Julia
Direct Derivatization in Dried Blood Spots for Oxidized and Reduced Glutathione Quantification in Newborns
title Direct Derivatization in Dried Blood Spots for Oxidized and Reduced Glutathione Quantification in Newborns
title_full Direct Derivatization in Dried Blood Spots for Oxidized and Reduced Glutathione Quantification in Newborns
title_fullStr Direct Derivatization in Dried Blood Spots for Oxidized and Reduced Glutathione Quantification in Newborns
title_full_unstemmed Direct Derivatization in Dried Blood Spots for Oxidized and Reduced Glutathione Quantification in Newborns
title_short Direct Derivatization in Dried Blood Spots for Oxidized and Reduced Glutathione Quantification in Newborns
title_sort direct derivatization in dried blood spots for oxidized and reduced glutathione quantification in newborns
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9219658/
https://www.ncbi.nlm.nih.gov/pubmed/35740062
http://dx.doi.org/10.3390/antiox11061165
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