Quantification of apixaban in human plasma using ultra performance liquid chromatography coupled with tandem mass spectrometry
10.12793/tcp.2019.27.1.33
- Author:
Hyeon Cheol JEONG
1
;
Tae Eun KIM
;
Kwang Hee SHIN
Author Information
1. College of Pharmacy, Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu 41566, Korea. kshin@knu.ac.kr
- Publication Type:Original Article
- Keywords:
Anticoagulant;
Apixaban;
Bioanalytical method;
UPLC-MS/MS
- MeSH:
Ammonium Compounds;
Anticoagulants;
Calibration;
Chromatography, Liquid;
Factor Xa;
Healthy Volunteers;
Hemorrhage;
Humans;
Mass Spectrometry;
Methanol;
Methods;
Plasma;
Stroke;
Tandem Mass Spectrometry
- From:Translational and Clinical Pharmacology
2019;27(1):33-41
- CountryRepublic of Korea
- Language:English
-
Abstract:
Apixaban, an inhibitor of direct factor Xa, is used for the treatment of venous thromboembolic events or prevention of stroke. Unlike many other anticoagulant agents, it does not need periodic monitoring. However, monitoring is still required to determine the risk of bleeding due to overdose or surgery. Usually, apixaban concentrations are indirectly quantified using an anti-factor Xa assay. However, this method has a relatively narrow analytical concentration range, poor selectivity, and requires an external calibrator. Therefore, the goal of current study was to establish an analytical method for determining plasma levels of apixaban using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). To this end, apixaban was separated using 2.5 mM ammonium formate (pH 3.0) (A) and 100% methanol containing 0.1% formic acid (B) using the gradient method with a Thermo hypersil GOLD column. The mass detector condition was optimized using the electrospray ionization (ESI) positive mode for apixaban quantification. The developed method showed sufficient linearity (coefficient of determination [r² ≥ 0.997]) at calibration curve ranges. The percentage (%) changes in accuracy, precision, and all stability tests were within 15% of the nominal concentration. Apixaban concentration in plasma from healthy volunteers was quantified using the developed method. The mean maximum plasma concentration (C(max)) was 371.57 ng/mL, and the median time to achieve the C(max) (T(max)) was 4 h after administration of 10 mg apixaban alone. Although the results showed low extraction efficiency (~16%), the reproducibility (% change was within 15% of nominal concentration) was reliable. Therefore, the developed method could be used for clinical pharmacokinetic studies.
