Development and validation of an LC-MS/MS method for the determination of a novel thienoquinolin urea transporter inhibitor PU-48 in rat plasma and its application to a pharmacokinetic study

  • Biomed Chromatogr. 2018 Apr;32(4). doi: 10.1002/bmc.4157.
Zhi-Yuan Zhang  1 ,  Xin Wang  1 ,  Dan Liu  2 ,  Hua Zhang  3 ,  Qiang Zhang  3 ,  Ying-Yuan Lu  1 ,  Pu Li  1 ,  Ya-Qing Lou  1 ,  Bao-Xue Yang  1 ,  Chuang Lu  4 ,  Ya-Xin Lou  2 ,  Guo-Liang Zhang  1
Affiliations
  • 1. Department of Pharmacology, School of Basic Medical Sciences, Peking University, Beijing, China.
  • 2. Proteomics Laboratory, Medical and Health Analysis Center, Peking University, Beijing, China.
  • 3. Department of Pharmaceutics, School of Pharmaceutical Sciences, Peking University, Beijing, China.
  • 4. Department of Drug Metabolism and Pharmacokinetics, Biogen, Cambridge, Massachusetts, USA.
PMID: 29193233 DOI: 10.1002/bmc.4157
Abstract

A specific, sensitive and stable high-performance liquid chromatographic-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantitative determination of methyl 3-amino-6-methoxythieno [2,3-b]quinoline-2-carboxylate (PU-48), a novel diuretic thienoquinolin Urea Transporter Inhibitor in rat plasma. In this method, the chromatographic separation of PU-48 was achieved with a reversed-phase C18 column (100 × 2.1 mm, 3 μm) at 35°C. The mobile phase consisted of acetonitrile and water with 0.05% formic acid added with a gradient elution at flow rate of 0.3 mL/min. Samples were detected with the triple-quadrupole tandem mass spectrometer with multiple reaction monitoring mode via electrospray ionization source in positive mode. The retention time were 6.2 min for PU-48 and 7.2 min for megestrol acetate (internal standard, IS). The monitored ion transitions were mass-to-charge ratio (m/z) 289.1 → 229.2 for PU-48 and m/z 385.3 → 267.1 for the internal standard. The calibration curve for PU-48 was linear over the concentration range of 0.1-1000 ng/mL (r2 > 0.99), and the lower limit of quantitation was 0.1 ng/mL. The precision, accuracy and stability of the method were validated adequately. The developed and validated method was successfully applied to the pharmacokinetic study of PU-48 in rats.

Keywords
liquid chromatography-tandem mass spectrometry (LC-MS/MS); methyl 3-amino-6-methoxythieno [2,3-b] quinolone-2-carboxylate (PU-48); pharmacokinetics; rat; thienoquinolin urea transporter inhibitor.
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