Hilic-ms/ms analysis of adenosine in patient blood

Abstract

Adenosine is a purine ribonucleoside with important roles in various physiological pro-cesses. A number of studies have indicated the importance of adenosine in cardiovascular diseases including syncope; however, the accurate determination of adenosine in human blood is challenging due to the molecule’s instability. In the present study, we report a simple method for the pre-treatment of blood samples and the development of a fast and efficient hydrophilic interaction chromatographic tandem mass spectrometry method for the analysis of adenosine in patient blood. During collec-tion, samples were mixed directly with a solvent mixture containing 95% acetonitrile and 10 mM ammonium formate in a Vacutainer tube, resulting in successful prevention of adenosine metabolic processes and direct blood sample deproteinization. The method was validated according to bioana-lytical industry guidelines and found to be accurate, repeatable, specific and sensitive with LLOQ 0.005 µg/mL, thus allowing its application in the analysis of real clinical samples.