Determination of Apixaban in human plasma by UPLC-MS/MS
OBJECTIVE To determine Apixaban in human plasma by UPLC-MS/MS.METHODS The samples were treated by protein precipitation with acetonitrile.Apixaban was separated on a Waters Xbridge C18(50 mm ×2.1 mm,3.5 μm)chromatographic column using gradient elution with the mobile phase consisting of 5％acetonitrile(A)and 95％acetonitrile(B)solution(containing 0.1％formic acid).The flow rate was set at 0.4 mL·min-1,the column temperature was set at 40 ℃,and the injection volume was 2 μL.The detection of the analytes was performed on electrospray ionization in positive mode by multiple monitoring with the transition of m/z 460.3→443.3(Apixaban)and m/z 464.4→447.3(13 C Apixaban-d3,IS).RESULTS A good linearity was observed for the Apixaban in the range of 0.5-400 ng·mL-1(r=0.9995),the lower limit of quantification was 0.5ng·mL-1,the recovery of Apixaban was ≥99.22％,and the intra-day and inter-day RSD of quality-control samples were ≤4.53％.The stability of whole blood and plasma samples met the requirements of biological sample analysis.CONCLUSION The method is easy,rapid,accurate and sensitive.It is suitable for the determination of Apixaban concentration in human plasma,and can be used to study the pharmacokinetics and bioequivalence of Apixaban tablets in healthy human subjects.
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