Direct oral anticoagulants (DOACs) have been commonly used for the treatment of venous thromboembolism and for the prevention of stroke in patients with atrial fibrillation. Despite not being initially recommended, monitoring DOACs plasma concentrations is now recognized as essential in emergency situations and in special populations. Moreover, the inter-individual variability found in real studies as well as the high reported non-adherence are corroborating the importance of determining the individual relationship between administered doses, plasma concentrations and pharmacological effects. Therefore, accurate but user-friendly bioanalytical techniques are required to monitor DOACs plasma concentrations in routine clinical practice and phase IV clinical trials. Herein, a fast and simple high performance liquid chromatography (HPLC) method coupled to diode array detection (DAD) was developed, validated and applied to quantify the four currently marketed DOACs (apixaban, edoxaban, dabigatran and rivaroxaban). Sample preparation was performed by solid phase extraction followed by evaporation and concentration of the analytes. Chromatographic separation was accomplished within 6 min on a reversed-phase column (octadecyl-silica packing material; 55 mm × 4 mm, 3 μm particle size), applying a mobile phase composed of an aqueous solution of formic acid (0.1 %, v/v) and acetonitrile, pumped with a gradient elution at 30 °C. The proposed method was linear (r2 ≥ 0.993) within the concentration ranges of 0.017-5.28 μg mL-1, 0.066-5.28 μg mL-1, 0.033-5.28 μg mL-1 and 0.017-5.28 μg mL-1 for apixaban, dabigatran, edoxaban and rivaroxaban, respectively, all of them including the expected range of therapeutic concentrations. Overall, intra- and inter-day trueness of quality control samples, including at the lower limit of quantification (LLOQ), varied between -12.98 to 5.79 %, while imprecision was lower than 16.43 %, supporting that the method is accurate and precise in accordance to international guidelines. Recovery and stability were also assessed and allowed the method to be applied in clinical practice, during therapeutic drug monitoring.

中文翻译：

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同时测定人血浆中阿哌沙班，达比加群，依多沙班和利伐沙班的新型HPLC-DAD方法的开发，验证和应用。

直接口服抗凝剂（DOAC）已普遍用于治疗静脉血栓栓塞和预防房颤患者的中风。尽管最初没有建议，但在紧急情况和特殊人群中，监测DOAC的血浆浓度现已被认为是必不可少的。此外，在实际研究中发现的个体间差异以及大量报道的不依从性，正证实了确定给药剂量，血浆浓度和药理作用之间的个体关系的重要性。因此，在常规临床实践和IV期临床试验中，需要准确而用户友好的生物分析技术来监测DOAC的血浆浓度。在这里 快速，简单的高效液相色谱（HPLC）方法与二极管阵列检测（DAD）结合开发，验证并应用于定量目前市售的四种DOAC（阿哌沙班，依多沙班，达比加群和利伐沙班）。通过固相萃取，然后蒸发和浓缩分析物进行样品制备。使用反相色谱柱（十八烷基硅胶填料； 55 mm×4 mm，粒径3μm），在6分钟内完成色谱分离，应用由甲酸水溶液（0.1％，v / v）和乙腈，在30°C下梯度洗脱。对于阿哌沙班，达比加群，在0.017-5.28μgmL-1、0.066-5.28μgmL-1、0.033-5.28μgmL-1和0.017-5.28μgmL-1的浓度范围内，所提出的方法是线性的（r2≥0.993） ，edoxaban和rivaroxaban分别包括预期的治疗浓度范围。总体上讲，质量控制样品的日内和日间真实性（包括定量下限（LLOQ））在-12.98％至5.79％之间，而不精确度则低于16.43％，这表明该方法在根据国际准则。还评估了恢复性和稳定性，并使该方法可用于治疗药物监测期间的临床实践。支持根据国际准则该方法的准确性和精确性。还评估了恢复性和稳定性，并使该方法可用于治疗药物监测期间的临床实践。支持根据国际准则该方法的准确性和精确性。还评估了恢复性和稳定性，并使该方法可用于治疗药物监测期间的临床实践。