Pasupuleti Raghavendra Rao, Tsai Pei-Chien, Ponnusamy Vinoth Kumar, Pugazhendhi Arivalagan
Department of Medicinal and Applied Chemistry, Kaohsiung Medical University (KMU), Kaohsiung City, 807, Taiwan.
Research Center for Environmental Medicine, Kaohsiung Medical University (KMU), Kaohsiung City, 807, Taiwan.
Process Biochem. 2021 Mar;102:150-156. doi: 10.1016/j.procbio.2020.12.014. Epub 2020 Dec 25.
To tackle the harmful consequences of the widespread COVID-19 pandemic, a broad-spectrum anti-viral drug remdesivir (RDV) has gained the utmost attention recently due to its promising application in treating COVID-19 patients. However, a fast and sensitive analytical methodology is important to monitor RDV drug profile in human plasma for pharmacokinetics (PK) and therapeutic drug monitoring (TDM). In this study, we demonstrate an improved vortex-assisted salt-induced liquid-liquid microextraction (VA-SI-LLME) technique coupled with UHPLC-PDA and UHPLC-MS/MS for rapid determination of RDV in human plasma. This technique involves simple one-step protein precipitation with hydrochloric acid and subsequent extraction with acetonitrile for analysis. Under the optimal VA-SI-LLME conditions (500 μL of acetonitrile with 2.5 g ammonium sulfate under 2 min vortex extraction), method validation results indicated an excellent correlation coefficient of 0.9969 for UHPLC-PDA (monitored at 254 nm) and 0.9990 for UHPLC-MS/MS (monitored at electrospray ionization with + ion mode transitions of 603.1→ 402.20 and 603.1→ 199.90). The detection and quantification limits were 1.5 and 5 ng/mL for UHPLC/PDA, and 0.3 and 1 ng/mL for UHPLC-MS/MS, respectively. The developed method showed excellent extraction recoveries between 90.79-116.74 % and 85.68-101.34 % with intraday and interday precision ≤ 9.59 for both methods. These results proved that the developed method is a simple, fast, and sensitive analytical method that can be applied as a standard analytical tool for PK and TDM studies of RDV in clinical trials during the current worldwide outbreak.
为应对广泛传播的新冠疫情的有害后果，一种广谱抗病毒药物瑞德西韦（RDV）因其在治疗新冠患者方面的应用前景最近受到了极大关注。然而，一种快速且灵敏的分析方法对于监测人血浆中瑞德西韦的药物特征以进行药代动力学（PK）和治疗药物监测（TDM）至关重要。在本研究中，我们展示了一种改进的涡旋辅助盐诱导液液微萃取（VA-SI-LLME）技术，该技术与超高效液相色谱-光电二极管阵列检测（UHPLC-PDA）和超高效液相色谱-串联质谱（UHPLC-MS/MS）联用，用于快速测定人血浆中的瑞德西韦。该技术包括用盐酸进行简单的一步蛋白沉淀，随后用乙腈萃取进行分析。在最佳的VA-SI-LLME条件下（500 μL乙腈与2.5 g硫酸铵在2分钟涡旋萃取下），方法验证结果表明，UHPLC-PDA（在254 nm处监测）的相关系数为0.9969，UHPLC-MS/MS（在电喷雾电离正离子模式下监测，跃迁为603.1→402.20和603.1→199.90）的相关系数为0.9990。UHPLC/PDA的检测限和定量限分别为1.5和5 ng/mL，UHPLC-MS/MS的检测限和定量限分别为0.3和1 ng/mL。所开发的方法显示出优异的萃取回收率，日内和日间精密度≤9.59%，两种方法的回收率分别在90.79 - 116.74%和85.68 - 101.34%之间。这些结果证明，所开发的方法是一种简单、快速且灵敏的分析方法，可作为当前全球疫情期间临床试验中瑞德西韦PK和TDM研究的标准分析工具。
