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Electromembrane Extraction of Highly Polar Compounds: Analysis of Cardiovascular Biomarkers in Plasma

Cardiovascular diseases (CVDs) represent a major concern in today’s society, with more than 17.5 million deaths reported annually worldwide. Recently, five metabolites related to the gut metabolism of phospholipids were identified as promising predictive biomarker candidates for CVD. Validation of t...

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Autores principales: Drouin, Nicolas, Kloots, Tim, Schappler, Julie, Rudaz, Serge, Kohler, Isabelle, Harms, Amy, Wilhelmus Lindenburg, Petrus, Hankemeier, Thomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022788/
https://www.ncbi.nlm.nih.gov/pubmed/31861366
http://dx.doi.org/10.3390/metabo10010004
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author Drouin, Nicolas
Kloots, Tim
Schappler, Julie
Rudaz, Serge
Kohler, Isabelle
Harms, Amy
Wilhelmus Lindenburg, Petrus
Hankemeier, Thomas
author_facet Drouin, Nicolas
Kloots, Tim
Schappler, Julie
Rudaz, Serge
Kohler, Isabelle
Harms, Amy
Wilhelmus Lindenburg, Petrus
Hankemeier, Thomas
author_sort Drouin, Nicolas
collection PubMed
description Cardiovascular diseases (CVDs) represent a major concern in today’s society, with more than 17.5 million deaths reported annually worldwide. Recently, five metabolites related to the gut metabolism of phospholipids were identified as promising predictive biomarker candidates for CVD. Validation of those biomarker candidates is crucial for applications to the clinic, showing the need for high-throughput analysis of large numbers of samples. These five compounds, trimethylamine N-oxide (TMAO), choline, betaine, l-carnitine, and deoxy-l-carnitine (4-trimethylammoniobutanoic acid), are highly polar compounds and show poor retention on conventional reversed phase chromatography, which can lead to strong matrix effects when using mass spectrometry detection, especially when high-throughput analysis approaches are used with limited separation of analytes from interferences. In order to reduce the potential matrix effects, we propose a novel fast parallel electromembrane extraction (Pa-EME) method for the analysis of these metabolites in plasma samples. The evaluation of Pa-EME parameters was performed using multi segment injection–capillary electrophoresis–mass spectrometry (MSI-CE-MS). Recoveries up to 100% were achieved, with variability as low as 2%. Overall, this study highlights the necessity of protein precipitation prior to EME for the extraction of highly polar compounds. The developed Pa-EME method was evaluated in terms of concentration range and response function, as well as matrix effects using fast-LC-MS/MS. Finally, the developed workflow was compared to conventional sample pre-treatment, i.e., protein precipitation using methanol, and fast-LC-MS/MS. Data show very strong correlations between both workflows, highlighting the great potential of Pa-EME for high-throughput biological applications.
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spelling pubmed-70227882020-03-11 Electromembrane Extraction of Highly Polar Compounds: Analysis of Cardiovascular Biomarkers in Plasma Drouin, Nicolas Kloots, Tim Schappler, Julie Rudaz, Serge Kohler, Isabelle Harms, Amy Wilhelmus Lindenburg, Petrus Hankemeier, Thomas Metabolites Article Cardiovascular diseases (CVDs) represent a major concern in today’s society, with more than 17.5 million deaths reported annually worldwide. Recently, five metabolites related to the gut metabolism of phospholipids were identified as promising predictive biomarker candidates for CVD. Validation of those biomarker candidates is crucial for applications to the clinic, showing the need for high-throughput analysis of large numbers of samples. These five compounds, trimethylamine N-oxide (TMAO), choline, betaine, l-carnitine, and deoxy-l-carnitine (4-trimethylammoniobutanoic acid), are highly polar compounds and show poor retention on conventional reversed phase chromatography, which can lead to strong matrix effects when using mass spectrometry detection, especially when high-throughput analysis approaches are used with limited separation of analytes from interferences. In order to reduce the potential matrix effects, we propose a novel fast parallel electromembrane extraction (Pa-EME) method for the analysis of these metabolites in plasma samples. The evaluation of Pa-EME parameters was performed using multi segment injection–capillary electrophoresis–mass spectrometry (MSI-CE-MS). Recoveries up to 100% were achieved, with variability as low as 2%. Overall, this study highlights the necessity of protein precipitation prior to EME for the extraction of highly polar compounds. The developed Pa-EME method was evaluated in terms of concentration range and response function, as well as matrix effects using fast-LC-MS/MS. Finally, the developed workflow was compared to conventional sample pre-treatment, i.e., protein precipitation using methanol, and fast-LC-MS/MS. Data show very strong correlations between both workflows, highlighting the great potential of Pa-EME for high-throughput biological applications. MDPI 2019-12-18 /pmc/articles/PMC7022788/ /pubmed/31861366 http://dx.doi.org/10.3390/metabo10010004 Text en © 2019 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Drouin, Nicolas
Kloots, Tim
Schappler, Julie
Rudaz, Serge
Kohler, Isabelle
Harms, Amy
Wilhelmus Lindenburg, Petrus
Hankemeier, Thomas
Electromembrane Extraction of Highly Polar Compounds: Analysis of Cardiovascular Biomarkers in Plasma
title Electromembrane Extraction of Highly Polar Compounds: Analysis of Cardiovascular Biomarkers in Plasma
title_full Electromembrane Extraction of Highly Polar Compounds: Analysis of Cardiovascular Biomarkers in Plasma
title_fullStr Electromembrane Extraction of Highly Polar Compounds: Analysis of Cardiovascular Biomarkers in Plasma
title_full_unstemmed Electromembrane Extraction of Highly Polar Compounds: Analysis of Cardiovascular Biomarkers in Plasma
title_short Electromembrane Extraction of Highly Polar Compounds: Analysis of Cardiovascular Biomarkers in Plasma
title_sort electromembrane extraction of highly polar compounds: analysis of cardiovascular biomarkers in plasma
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022788/
https://www.ncbi.nlm.nih.gov/pubmed/31861366
http://dx.doi.org/10.3390/metabo10010004
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