A liquid chromatography-tandem mass spectrometry method for the determination of apixaban in human plasma and its application to pharmacokinetics studies in the Indian population.

Khurshid Shaikh, Ashish Mungantiwar, Supriya Halde, Nancy Pandita

Analytical methods : advancing methods and applications 2021 Sep 07

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Apixaban is a novel oral anticoagulant intended to treat and prevent blood clots and to prevent strokes in patients with nonvalvular atrial fibrillation. The development and validation of a fast, selective, accurate, and precise method using high-performance liquid chromatography tandem mass spectrometry is described for the estimation of apixaban in human plasma, with apixaban 13CD3 as an internal standard (IS). Using a reverse phase Gemini C18 column (50 mm × 4.6 mm, 3 μm) and a mixture of acetonitrile (2 mM) and ammonium formate buffer (50 : 50 v/v) as the mobile phase, chromatographic separation was achieved following extraction via a solid-phase extraction process. To track multiple reaction monitoring transitions set at 460/443 (m/z) and 464/447 (m/z) for apixaban and apixaban 13CD3, respectively, liquid chromatography coupled with triple quadrupole mass spectrometry was employed. A concentration linearity range between 1.01 and 280.00 ng mL-1 was validated with regression ≥0.99, and the method was successfully applied to apixaban pharmacokinetics analysis. At a flow rate of 1.0 mL min-1, the run time was around 1.8 min, which is short. With an extraction recovery of >73% for both apixaban and apixaban 13CD3, the method was sensitive, with a limit of quantitation of 1.01 ng mL-1. The inter-day/between-run precision ranged from 1.21% to 3.21%, while the accuracy ranged from 96.5% to 102%. For pharmacokinetics analysis, the validated method was applied. The percentage difference between findings from samples that were reanalyzed and samples that were initially analyzed was within ±20%. With high-quality assay specificity and accuracy in relation to apixaban analysis in human plasma under the experimental conditions used, the method provided is accurate.

Citation

Khurshid Shaikh, Ashish Mungantiwar, Supriya Halde, Nancy Pandita. A liquid chromatography-tandem mass spectrometry method for the determination of apixaban in human plasma and its application to pharmacokinetics studies in the Indian population. Analytical methods : advancing methods and applications. 2021 Sep 07;13(33):3693-3704