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

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 t...

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Bibliographic Details
Published in:Pharmaceuticals (Basel, Switzerland) Switzerland), 2024-03, Vol.17 (3), p.345
Main Authors: Šofranko, Jakub, Mitro, Peter, Lazúrová, Zora, Péč, Martin Jozef, Bolek, Tomáš, Péčová, Renata, Dohál, Matúš, Samoš, Matej, Murín, Radovan
Format: Article
Language:English
Subjects:
Accuracy
Adenosine
blood
Calibration
Chromatography
Health aspects
hypoxanthine
inosine
LC–MS
Liquid chromatography
Mass spectrometry
Measurement
Physiology
Proteins
Retention
Scientific imaging
Signal transduction
Uric acid
xanthine
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Summary: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 beat 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 collection, a blood sample mixed with acetonitrile solution that is either enriched with C-labeled adenosine or a newly generated mixture is transferred into the tubes containing the defined amount of C-labeled adenosine. The C-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 for quantification of a few catabolites of adenosine, i.e., inosine, hypoxanthine, and xanthine. Our current setup did not allow for the detection or quantifying of uric acid, which is the final product of adenosine catabolism. This advancement provides an 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.
ISSN:1424-8247
1424-8247
DOI:10.3390/ph17030345