Department of Medicine, University of Otago, Christchurch; and.
Toxicology, Department of Specialist Biochemistry, Canterbury Health Laboratories, Christchurch, New Zealand.
Ther Drug Monit. 2020 Jun;42(3):473-480. doi: 10.1097/FTD.0000000000000744.
Pharmacokinetic studies and therapeutic drug monitoring of anticoagulants require a simple, rapid, and reliable analytical method for monitoring plasma concentrations. The aims of the current work were to develop and validate a liquid chromatography/tandem mass spectrometry method for the simultaneous determination of 3 direct oral anticoagulants (dabigatran, rivaroxaban, and apixaban) in human plasma that is suitable for pharmacokinetic studies and routine therapeutic drug monitoring in busy hospital laboratories.
This method included a hydrolysis step to account for the active acylglucuronide metabolites of dabigatran that demonstrate an equivalent anticoagulant effect as dabigatran. After hydrolysis, a simple one-step protein precipitation was used for sample preparation. Total dabigatran (the sum of free dabigatran and the contribution from dabigatran acylglucuronides), rivaroxaban, and apixaban, and their corresponding isotopically labeled internal standards were resolved on a C18(2) column. All compounds were detected using electrospray ionization liquid chromatography/tandem mass spectrometry in the positive mode.
For all 3 anticoagulants, standard curves were linear over the concentration range of 1.0-1000 mcg/L (r > 0.99), bias was < ±10%, and intraday and interday coefficients of variation (imprecision) were <10%. The limit of quantification was 1.0 mcg/L. For all 3 anticoagulants and corresponding isotopically labeled internal standards, the absolute recoveries were similar and consistent, with mean values of 93%-102%. No significant matrix effects were observed.
This method is simple, rapid, robust, and reliable and can be used to analyze the plasma concentrations of the drugs in patients on dabigatran or rivaroxaban therapy.
抗凝药物的药代动力学研究和治疗药物监测需要一种简单、快速、可靠的分析方法来监测血浆浓度。本研究旨在开发和验证一种适用于药代动力学研究和繁忙医院实验室常规治疗药物监测的同时测定人血浆中 3 种直接口服抗凝剂（达比加群、利伐沙班和阿哌沙班）的液相色谱/串联质谱法。
该方法包括水解步骤，以解释达比加群的活性酰基葡萄糖醛酸代谢物，这些代谢物具有与达比加群相当的抗凝作用。水解后，采用简单的一步蛋白沉淀法进行样品制备。总达比加群（游离达比加群和达比加群酰基葡萄糖醛酸代谢物的总和）、利伐沙班和阿哌沙班及其相应的同位素标记内标物在 C18（2）柱上得到分离。所有化合物均采用电喷雾液相色谱/串联质谱法在正离子模式下检测。
对于所有 3 种抗凝剂，标准曲线在 1.0-1000 mcg/L 浓度范围内呈线性（r > 0.99），偏差< ±10%，日内和日间变异系数（精密度）<10%。定量下限为 1.0 mcg/L。对于所有 3 种抗凝剂和相应的同位素标记内标物，绝对回收率相似且一致，平均值为 93%-102%。未观察到明显的基质效应。
该方法简单、快速、稳健、可靠，可用于分析达比加群或利伐沙班治疗患者的药物血浆浓度。
