Development and validation of a hydrophilic interaction liquid chromatography‐tandem mass spectrometry method for the quantification of regadenoson in human plasma and its pharmacokinetic application

• Published in: Journal of separation science Vol. 45; no. 6; pp. 1146 - 1152
• Main Authors: Wang, Dun‐Jian, Wang, Da‐Wei, Fang, Qiu‐Chen, Shen, Ye, Zeng, Nv‐Jin, Yang, Yan‐Ling, Zhang, Hong‐Wen, Wang, Yong‐Qing, Sun, Lu‐Ning
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 01.03.2022
• Subjects: Acetonitrile; Adenosine; Ammonium acetate; Blood plasma; Cardiac stress tests; Chromatography, High Pressure Liquid; Chromatography, Liquid - methods; Humans; hydrophilic interaction liquid chromatography; Hydrophobic and Hydrophilic Interactions; Ions; Liquid chromatography; Mass spectrometry; Pharmacokinetics; Pharmacology; Purines; Pyrazoles; Radioisotopes; regadenoson, tandem mass spectrometry; Reproducibility of Results; Scientific imaging; Spectroscopy; Tandem Mass Spectrometry - methods; pharmacokinetics; regadenoson, tandem mass spectrometry; hydrophilic interaction liquid chromatography
• ISSN: 1615-9306; 1615-9314
• DOI: 10.1002/jssc.202100756

Regadenoson, the first selective adenosine A2A receptor agonist, is used to perform exercise stress test during radionuclide myocardial perfusion imaging. To detect the concentration of regadenoson in human plasma, a simple, fast, and sensitive tandem mass spectrometry method was established herein. Acetonitrile was used as a protein precipitation agent. Chromatographic separation was completed in 6.5 min using a BEH HILIC column (50 × 2.1 mm, 1.7 μm). The mobile phase consisted of 10 mmol/L ammonium acetate/acetonitrile (gradient elution). To quantify regadenoson and regadenoson‐d3, an API 4000 mass spectrometry in multiple reaction monitoring mode with transitions of 391.3→259.2 and 394.3→262.2, respectively, was utilized. The calibration curve was linear in the range of 0.100–50.0 μg/L, and the intrabatch and interbatch precisions were <9.7% and <13.0%, respectively, and the accuracy was 2.0–6.9%. There was no apparent matrix effect for regadenoson or regadenoson‐d3. The developed method was used to study the pharmacokinetic characteristics of regadenoson in healthy Chinese subjects.