Determination of Tropifexor in Beagle Dog Plasma by UPLC-MS/MS and Its Application in Pharmacokinetics

Abstract

The primary objective of this study was to develop and validate an efficient and accurate ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) approach as a means to detect tropifexor plasma concentrations in beagle dogs and to study its pharmacokinetic profile in beagle dogs. The chromatographic separation of tropifexor and oprozomib (internal standard, ISTD) on the column, with the addition of acetonitrile for rapid precipitation and protein extraction, was achieved with 0.1% formic acid aqueous solution-acetonitrile for the mobile phase. A Xevo TQ-S triple quadrupole tandem mass spectrometer, under the selective reaction monitoring (SRM) mode, for the determination of the concentrations in the positive ion mode. The mass transfer pairs of tropifexor and oprozomib (ISTD) were m/z 604.08 ⟶ 228.03 and m/z 533.18 ⟶ 199.01, respectively. The profile displayed well linearity with calibration curves for tropifexor and oprozomib (ISTD) ranging from 1.0 to 200 ng/mL. In parallel, the lower limit of quantification (LLOQ) value for tropifexor could be measured with the aid of this novel technique at 1.0 ng/mL. In addition, the scope of intraday and interday for analyte accuracy was between −4.86% and 1.16%, with a precision of <7.31%. The recoveries of the analytes were >88.13% and were free of significant matrix effects. The stability met the requirements for the quantification of plasma samples under various conditions. Finally, the pharmacokinetic profile of tropifexor in beagle dog plasma following oral administration of 0.33 mg/kg tropifexor was determined by using the method facilitated in this work.

A Reliable and Effective UPLC-MS/MS Method for the Determination of Oprozomib in Rat Plasma

“…This analysis method followed FDA guidelines for the validation of bioanalytical tests, and the method validation procedures including the calibration curve, stability, matrix effect, LLOQ, precision, and accuracy were fully conducted [ 12 ].…”

“…Method Validation. Tis analysis method followed FDA guidelines for the validation of bioanalytical tests, and the method validation procedures including the calibration curve, stability, matrix efect, LLOQ, precision, and accuracy were fully conducted [12]. In this experiment, in order to detect the selectivity of the method, blank plasma samples from six diferent rats were analyzed by chromatography to evaluate whether there was endogenous interference at the retention times of both the analyte and IS in the method.…”