Bioequivalence Assessment of Apixaban in Human Plasma: Analytical Method Validation and Pharmacokinetic Profiling Using LC -MS/MS

The accurate quantification of Apixaban in human plasma is essential for bioequivalence and pharmacokinetic studies. This study presents a validated analytical method for the determination of Apixaban using Liquid Chromatography –Mass Spectrometry/Mass Spectrometry (LC -MS/MS). The method employs a Quattro Micromass spectrometer in positive electrospray ionization mode and an Agilent Zorbax SB-C۱۸ column for chromatographic separation. Calibration was performed within a concentration range of ۱.۲۵ to ۳۰۰ ppb, with a weighted (۱/X) regression model demonstrating excellent linearity (R² > ۰.۹۹). Method validation followed ICH M۱۰ guidelines, evaluating specificity, accuracy, precision, matrix effects, and recovery. Specificity testing showed no significant interference from blank plasma matrices. Intra-day and inter-day precision (RSD%) remained below ۱۵%, confirming method robustness. The mean recovery of Apixaban from plasma samples was >۸۵%, with minimal matrix effects. Bioequivalence was assessed in human subjects, comparing pharmacokinetic parameters of test and reference formulations. Plasma concentrations were quantified at multiple time points post-administration, and pharmacokinetic profiles were constructed. The study confirmed the bioequivalence of the formulations based on C_max, AUC_۰-t, and AUC_۰-∞ values within the accepted regulatory range (۸۰-۱۲۵%). The validated LC-MS/MS method ensures precise and reproducible quantification of Apixaban in human plasma, supporting bioequivalence studies and regulatory submissions. The findings contribute to the advancement of analytical methodologies in clinical pharmacokinetics and generic drug development.

کلیدواژه ها:

Apixaban ، Bioequivalence ، LC-MS/MS ، Pharmacokinetics ، Analytical Method Validation ، Human Plasma ، Calibration Curve ، Drug Metabolism