Department of Bioequivalence, Research & Development Centre, Macleods Pharmaceuticals Limited, Plot No. 18, Street No. 9, MIDC, Andheri - (East), Mumbai 400093, India.
Anal Methods. 2021 Sep 7;13(33):3693-3704. doi: 10.1039/d1ay00837d. Epub 2021 Aug 1.
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 CD 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 CD, respectively, liquid chromatography coupled with triple quadrupole mass spectrometry was employed. A concentration linearity range between 1.01 and 280.00 ng mL 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, the run time was around 1.8 min, which is short. With an extraction recovery of >73% for both apixaban and apixaban CD, the method was sensitive, with a limit of quantitation of 1.01 ng mL. 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.
阿哌沙班是一种新型的口服抗凝剂，旨在治疗和预防血栓形成，并预防非瓣膜性心房颤动患者的中风。本文描述了一种使用高效液相色谱串联质谱法快速、选择性、准确和精密测定人血浆中阿哌沙班的方法，阿哌沙班 CD 作为内标（IS）。采用反相 Gemini C18 柱（50mm×4.6mm，3μm）和乙腈（2mM）和甲酸铵缓冲液（50：50v/v）的混合物作为流动相，通过固相萃取法进行提取，实现色谱分离。采用液相色谱-三重四极杆质谱法，分别跟踪阿哌沙班和阿哌沙班 CD 的多反应监测转换 460/443（m/z）和 464/447（m/z）。采用回归≥0.99验证了 1.01-280.00ng mL 之间的浓度线性范围，该方法成功应用于阿哌沙班的药代动力学分析。在 1.0mL min的流速下，运行时间约为 1.8 min，较短。阿哌沙班和阿哌沙班 CD 的提取回收率均>73%，方法灵敏，定量下限为 1.01ng mL。日内/日间精密度范围为 1.21%-3.21%，准确度范围为 96.5%-102%。在药代动力学分析中，应用了验证的方法。重新分析的样品和最初分析的样品之间的发现之间的百分比差异在±20%以内。在实验条件下，该方法对人血浆中阿哌沙班分析具有良好的特异性和准确性，结果准确。
