Bioequivalence Assessment of Apixaban in Human Plasma: Analytical Method Validation and Pharmacokinetic Profiling Using LC-MS/MS
کلمات کلیدی:
Apixaban, Bioequivalence, LC-MS/MS, Pharmacokineticsچکیده
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 Micro mass spectrometer in positive electrospray ionization mode and an Agilent Zorbax SB-C18 column for chromatographic separation. Calibration was performed within a concentration range of 1.25 to 300 ppb, with a weighted (1/X) regression model demonstrating excellent linearity (R² > 0.99). Method validation followed ICH M10 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 15%, confirming method robustness. The mean recovery of Apixaban from plasma samples was >85%, 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_0-t_, and AUC_0-∞ values within the accepted regulatory range (80-125%). 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.
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مراجع
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