Pharmacokinetics, bioavailability and metabolism of scopoletin in dog by ultra-high-performance liquid chromatography combined with linear ion trap-Orbitrap tandem mass spectrometry.

Abstract

A highly sensitive and selective method based on ultra-high-performance liquid chromatography combined with linear ion trap-Orbitrap tandem mass spectrometry (UHPLC-LTQ-Orbitrap-MS) has been developed and validated for the determination of scopoletin in dog plasma. The analyte was extracted from plasma samples using acetonitrile and separated on an Acquity UPLC BEH C18 column (50 × 2.1 mm, 1.7 μm) with 0.05% ammonium hydroxide and acetonitrile as mobile phase. The developed method was linear over the concentration range of 1-500 ng/mL, with a correlation coefficient >0.9988. The intra- and inter-day precisions (RSD) were <8.93% while the accuracy (RE) ranged from -6.50 to 8.12%. Extraction recovery, matrix effect and stability for dog plasma samples were within the required limits. The validated method has been successfully applied to investigate the pharmacokinetics and metabolism of scopoletin in dog plasma after intravenous (1 mg/kg) and oral (10, 25, 50 mg/kg) administration. The results revealed that (a) scopoletin showed short elimination half-life in dog; (b) its oral bioavailability was low (within the range of 5.69-7.08%); (c) scopoletin showed dose-independent pharmacokinetic profiles in dog plasma over the dose range of 10-50 mg/kg; and (d) glucuronidation was the predominant metabolic pathway in dog.