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Open AccessArticle

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

1
Institute of Pharmaceutical Sciences of Western Switzerland (ISPSO), University Medical Centre, 1206 Geneva, Switzerland
2
Division of Systems Biomedicine and Pharmacology, Leiden Academic Center for Drug Research, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands
3
Research Group Metabolomics, Leiden Centre for Applied Bioscience, University of Applied Sciences Leiden, Zernikedreef 11, 2333 CK Leiden, The Netherlands
*
Author to whom correspondence should be addressed.
Metabolites 2020, 10(1), 4; https://doi.org/10.3390/metabo10010004
Received: 15 November 2019 / Revised: 5 December 2019 / Accepted: 11 December 2019 / Published: 18 December 2019
(This article belongs to the Special Issue Sample Preparation in Metabolomics)
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. View Full-Text
Keywords: electromembrane extraction; cardiovascular disease; multi-segment injection; capillary electrophoresis–mass spectrometry; liquid chromatography–mass spectrometry electromembrane extraction; cardiovascular disease; multi-segment injection; capillary electrophoresis–mass spectrometry; liquid chromatography–mass spectrometry
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Drouin, N.; Kloots, T.; Schappler, J.; Rudaz, S.; Kohler, I.; Harms, A.; Wilhelmus Lindenburg, P.; Hankemeier, T. Electromembrane Extraction of Highly Polar Compounds: Analysis of Cardiovascular Biomarkers in Plasma. Metabolites 2020, 10, 4.

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