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Abstract

LC-MS/MS ASSAY FOR THE DETERMINATION OF APIXABAN IN HUMAN PLASMA: ITS APPLICATION TO A PHARMACOKINETIC STUDY

Seelam Sharath Chandra*, Rakesh Kumar Jat, Nageswara Rao Pilli, Premkumar Jangam and K. Soujanya

ABSTRACT

Introduction: Apixaban is a novel oral anticoagulant for the management of thromboembolic disease. In the present research work, a novel LC-MS/MS method was developed and validated for the determination of apixaban in human plasma. Method: Apixaban 13Cd3 was used as an internal standard (IS). A C18 column with isocratic mobile phase of 0.1% formic acid – methanol (15:85, v/v) used for the separation of extracted analyte. The flow rate was 0.75mL/min. The proposed linearity for apixaban was 1.02–301.3 ng/mL. Results: A total of five linearity curves were generated with quality control samples to calculate the precision and accuracy. Variety of validation experiments namely precision and accuracy, linearity, dilution integrity, recovery and matrix effect were well within the acceptablelimits. Also, the stability of analyte was extensively evaluated in plasma as well in extracted samples and results were met the acceptance criteria as per the regulatory guidelines. Conclusion: The present LC–MS/MS assay method is simple, rapid and sensitive for the determination of apixaban in human plasma. This is well suitable for pharmacokinetic or bioavailability/bioequivalence application.

Keywords: Apixaban, Human plasma, SPE, LC–MS/MS, Pharmacokinetics.


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