Department of Pharmacy, Beijing Chao-Yang Hospital, Capital Medical University, No.8 Gongti South Road, Chaoyang District, Beijing, 100020, People's Republic of China.
Anal Bioanal Chem. 2021 Sep;413(23):5811-5820. doi: 10.1007/s00216-021-03561-8. Epub 2021 Jul 24.
Remdesivir is a nucleotide analog prodrug that has received much attention since the outbreak of the COVID-19 pandemic in December 2019. GS-441524 (Nuc) is the active metabolite of remdesivir and plays a pivotal role in the clinical treatment of COVID-19. Here, a robust HPLC-MS/MS method was developed to determine Nuc concentrations in rat plasma samples after a one-step protein precipitation process. Chromatographic separation was accomplished on Waters XBrige C column (50 × 2.1 mm, 3.5 μm) under gradient elution conditions. Multiple reaction monitoring transitions in electrospray positive ion mode were m/z 292.2 → 163.2 for Nuc and 237.1 → 194.1 for the internal standard (carbamazepine). The quantitative analysis method was fully validated in line with the United States Food and Drug Administration guidelines. The linearity, accuracy and precision, matrix effect, recovery, and stability results met the requirements of the guidelines. Uncertainty of measurement and incurred sample reanalysis were analyzed to further ensure the robustness and reproducibility of the method. This optimized method was successfully applied in a rat pharmacokinetics study of remdesivir (intravenously administration, 5 mg kg). The method can act as a basis for further pharmacokinetic and clinical efficacy investigations in patients with COVID-19. Graphical abstract.
瑞德西韦是一种核苷酸类似物前药，自 2019 年 12 月 COVID-19 大流行爆发以来，受到了广泛关注。GS-441524（Nuc）是瑞德西韦的活性代谢物，在 COVID-19 的临床治疗中发挥着关键作用。在这里，开发了一种强大的 HPLC-MS/MS 方法，用于在一步蛋白沉淀过程后测定大鼠血浆样品中的 Nuc 浓度。在梯度洗脱条件下，色谱分离在 Waters XBridge C 柱（50×2.1mm，3.5μm）上完成。电喷雾正离子模式下的多重反应监测转换为 m/z 292.2→163.2 用于 Nuc 和 m/z 237.1→194.1 用于内标（卡马西平）。定量分析方法完全按照美国食品和药物管理局的指南进行了验证。线性、准确度和精密度、基质效应、回收率和稳定性结果均符合指南要求。测量不确定度和样品再分析的引入分析进一步确保了方法的稳健性和重现性。该优化方法成功应用于大鼠瑞德西韦（静脉注射，5mg/kg）的药代动力学研究。该方法可以作为进一步在 COVID-19 患者中进行药代动力学和临床疗效研究的基础。
