Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Application of Liquid Chromatography Coupled to Mass Spectrometry for Direct Estimation of the Total Levels of Adenosine and Its Catabolites in Human Blood

Jakub Šofranko
,
Peter Mitro
ORCID logo ,
Zora Lazúrová
,
Martin Jozef Péč
,
Tomáš Bolek
,
Renata Péčová
,
Matej Samoš
,
Radovan Murín
* ORCID logo
Version 1 : Received: 6 February 2024 / Approved: 6 February 2024 / Online: 6 February 2024 (09:48:20 CET)

A peer-reviewed article of this Preprint also exists.

Šofranko, J.; Mitro, P.; Lazúrová, Z.; Péč, M.J.; Bolek, T.; Péčová, R.; Dohál, M.; Samoš, M.; Murín, R. Application of Liquid Chromatography Coupled to Mass Spectrometry for Direct Estimation of the Total Levels of Adenosine and Its Catabolites in Human Blood. Pharmaceuticals 2024, 17, 345. Šofranko, J.; Mitro, P.; Lazúrová, Z.; Péč, M.J.; Bolek, T.; Péčová, R.; Dohál, M.; Samoš, M.; Murín, R. Application of Liquid Chromatography Coupled to Mass Spectrometry for Direct Estimation of the Total Levels of Adenosine and Its Catabolites in Human Blood. Pharmaceuticals 2024, 17, 345.

Abstract

Adenosine is a multifunctional nucleoside with several roles across various levels in organisms. Beyond its intracellular involvement in cellular metabolism, extracellular adenosine potently influences both physiological and pathological processes. In relation to its blood level, adenosine impacts the cardiovascular system, such as heart beating rate and vasodilation. To exploit the adenosine levels in the blood, we employed the liquid chromatography method coupled with mass spectrometry (LC-MS). Immediately after the collection, a blood sample mixed with acetonitrile solution that is either enriched with 13C-labeled adenosine or a newly generated mixture is transferred into the tubes containing the defined amount of 13C-labeled adenosine. The 13C-enriched isotopic adenosine is used as an internal standard, allowing for more accurate quantification of adenosine. This novel protocol for LC-MS-based estimation of adenosine delivers a rapid, highly sensitive, and reproducible means for quantitative estimation of total adenosine in blood. The method also allows quantification of a few catabolites of adenosine, i.e., inosine, hypoxanthine, and xanthine. This advancement provides the analytical tool that has the potential to enhance our understanding of adenosine's systemic impact and pave the way for further investigations into its intricate regulatory mechanisms.

Keywords

adenosine; inosine; xanthine; blood; LC-MS

Subject

Medicine and Pharmacology, Medicine and Pharmacology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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