Abstract:

In this study, a sensitive and rapid LC-MS/MS method was developed and validated to determine dabigatran in plasma of beagle dogs after oral administration of dabigatran etexilate nanosuspension (DABE-NS). The analytes (dabigatran) and sertraline hydrochloride (internal standard, IS) were separated on a Kromasil C₁₈ column using gradient elution consisting of methanol and formate buffer at a flow rate of 0.4 mL/min in 20 min. Detection and quantitation were carried out by multiple reaction monitoring following the transitions: m/z 472.17→289.07 and 305.98→275.00 for dabigatran and IS at positive ion mode, respectively. The calibration curves were linear from 1.0 to 500.0  ng/mL for dabigatran with r  = 0.9995. The accuracy of each analyte ranged from 94.8% to 107.1%, and the precision was within 6%. Besides, this method was successfully applied in the investigation of the pharmacokinetic profile of dabigatran in beagle dogs after oral administration ofDABE-NS. The maximum concentration and the areas under curves of dabigatranfor DABE-NS were significantly higher than those of control formulation, indicating improved oral absorption.

Key words: Dabigatran, Dabigatran etexilate, LC-MS/MS, Nanosuspension, Pharmacokinetics