Determination of duloxetine in human plasma with proven lack of influence of the major metabolite 4-hydroxyduloxetine

Abstract

Objectives: Minimizing the impact of major or unstable metabolites on the determination of a drug substance represents a leading task in the development and validation of bioanalytical methods. "Incurred samples reanalysis" provides relevant information too late; therefore, carefully selected tests on known metabolites should precede the pharmacokinetic studies.

Design and methods: This paper describes a simple and rapid HPLC-UV method for the determination of duloxetine, a potent serotonin and norepinephrine reuptake inhibitor, in the presence of its major metabolite, i.e. 4-hydroxyduloxetine glucuronide. Analyte and fluoxetine (internal standard) were extracted from human plasma by liquid-liquid extraction.

Results: No influence of the major metabolite was observed on the reliability of the new method. There was also lack of evidence of the major metabolite back-conversion to the parent drug substance. The validation demonstrated high precision of the new method. All validation parameters met the acceptance criteria of bioanalytical regulations.

Conclusions: The new method enabled the reliable determination of duloxetine in the presence of its major metabolite in the human plasma. The method might be applied to pharmacokinetic studies in humans, including bioequivalence and therapeutic drug monitoring.

Keywords: 4-Hydroxyduloxetine glucuronide; HPLC; Metabolite back-conversion; Pharmacokinetics; Validation.