METHOD DEVELOPMENT AND VALIDATION OF SELECTIVE AND HIGHLY SENSITIVE METHOD FOR DETERMINATION OF APIXABAN IN HUMAN PLASMA USING LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY

  • CHAITANYA KRISHNA ATMAKURI Faculty of Pharmacy, Pacific Academy of Higher Education and Research University, Udaipur, Rajasthan, India
  • CHEEPURUPALLI PRASAD Vignan Institute of Pharmaceutical Technology, Visakhapatnam, India
  • RAJARAM S. PATIL Director, Omacon Analytical Solutions LLP, Thane, India
  • KARUNAKRANTH DHARANI M. Pharm. Ph. D. Pharmaceutical Analysis, Mumbai, India

Abstract

Objective: The present research work aims to develop and validate a selective and highly sensitive method for the determination of apixaban in human plasma using liquid chromatography-tandem mass spectrometry (LC-MS/MS).


Methods: 200 µl of sodium heparin plasma samples were acidified and clean-up was performed by using solid-phase extraction (SPE). Apixaban 13C D3 was used as an internal standard (deuterated) to lower the relative matrix effects and a single step SPE was employed for sample clean up. 10 µl of SPE eluent was loaded onto Hypersil Beta Basic C18, 100×4.6 mm, 5 µ column for highly selective chromatographic separation using an isocratic mobile phase. 2 mmol ammonium acetate in water and acetonitrile were delivered by using a quaternary low-pressure gradient pump without premixing at a minimum flow rate of 0.50 ml/min.


Results: LC-MS/MS method was successfully developed and validated to demonstrate the lowest detection limit of 0.05 ng/ml and a linear dynamic range from 1-250 ng/ml with r2>0.99. Method development and validation results proved that the method is selective and highly sensitive for the determination of apixaban in human plasma using LC-MS/MS.


Conclusion: Current method can be applied for both therapeutic drug monitoring (TDM) and pharmacokinetic (PK) study analysis.

Keywords: Human Plasma, Liquid Chromatography-Tandem, Mass spectrometry

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ATMAKURI, C. K., C. PRASAD, R. S. PATIL, and K. DHARANI. “METHOD DEVELOPMENT AND VALIDATION OF SELECTIVE AND HIGHLY SENSITIVE METHOD FOR DETERMINATION OF APIXABAN IN HUMAN PLASMA USING LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 11, no. 8, July 2019, pp. 39-45, doi:10.22159/ijpps.2019v11i8.33948.
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