Simultaneous determination of adenosine triphosphate and its metabolites in human whole blood by RP-HPLC and UV-detection

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2008-03, Vol.864 (1), p.43-51
• Main Authors: Coolen, Erik J.C.M., Arts, Ilja C.W., Swennen, Els L.R., Bast, Aalt, Stuart, Martien A. Cohen, Dagnelie, Pieter C.
• Format: Article
• Language: English
• Subjects: Adenine Nucleotides - blood; Adenosine; Adenosine Triphosphate - blood; Analysis; Analytical, structural and metabolic biochemistry; ATP; Biological and medical sciences; Blood; Chromatography, High Pressure Liquid - methods; Drug Stability; Fundamental and applied biological sciences. Psychology; General pharmacology; HPLC; Humans; Medical sciences; Pharmacology. Drug treatments; Reproducibility of Results; Sensitivity and Specificity
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2008.01.033

To obtain insight in mechanisms of action of extracellular adenosine triphosphate (ATP) and adenosine, a simple HPLC method has been optimized and applied to investigate ATP metabolism in human whole blood ex vivo. This method provided good chromatographic resolution and peak shape for all eight compounds within a 19 min run time. The baseline was clean, the lower limit of quantification was below 0.3 μmol/L for all adenine nucleotides and the method demonstrated good linearity. Within-day precision ranged from 0.7 to 5.9% and between-days from 2.6 to 15.3%. Simplicity and simultaneous detection of ATP and its metabolites make this method suitable for clinical pharmacokinetic studies.