Quantitation of tizoxanide in multiple matrices to support cell culture, animal and human research.
Quantitation of tizoxanide in multiple matrices to support cell culture, animal and human research.
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DOI：
10.1101/2021.05.27.445500
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发表时间：
2021-05
期刊：
影响因子：
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通讯作者：
M. Neary;U. Arshad;Lee M. Tatham;H. Pertinez;H. Box;R. Rajoli;A. Valentijn;Joanne Sharp;S. Rannard;G. Biagini;P. Curley;A. Owen
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文献类型：
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作者：
M. Neary;U. Arshad;Lee M. Tatham;H. Pertinez;H. Box;R. Rajoli;A. Valentijn;Joanne Sharp;S. Rannard;G. Biagini;P. Curley;A. Owen
目前，硝唑尼特正被评估为SARS-CoV-2的候选治疗药物。与正在研究的许多其他候选药物不同，替唑尼特（硝唑尼特的活性代谢物）的血浆浓度在批准剂量给药后达到抗病毒水平，尽管预计在大多数患者中需要更高的剂量才能在给药间隔内维持这些浓度。本文描述了一种已根据美国食品药品监督管理局（FDA）指南进行验证的LC-MS/MS测定法。基本参数进行了评价，其中包括准确度，精密度和灵敏度。对人血浆、小鼠血浆和含不同浓度胎牛血清（FBS）的Dulbeccos改良Eagles培养基（DMEM）进行了试验验证。基质效应是色谱分析的一个有据可查的问题来源，可能影响分析过程的各个阶段，包括抑制或增强电离。因此，提出了一种稳健验证的LC-MS/MS分析方法，能够定量多种基质中的替唑尼特，基质效应的影响最小。此处提供的经验证的含量测定在15.6 ng/mL至1000 ng/mL范围内呈线性。准确度和精密度范围分别为102.2%和113.5%、100.1%和105.4%。本文所提出的检测方法在临床前和临床研究中均有应用，可用于促进硝唑尼特抗SARS-CoV-2应用的进一步研究。
Currently nitazoxanide is being assessed as a candidate therapeutic for SARS-CoV-2. Unlike many other candidates being investigated, tizoxanide (the active metabolite of nitazoxanide) plasma concentrations achieve antiviral levels after administration of the approved dose, although higher doses are expected to be needed to maintain these concentrations across the dosing interval in the majority of patients. Here an LC-MS/MS assay is described that has been validated in accordance with Food and Drug Administration (FDA) guidelines. Fundamental parameters have been evaluated, and these included accuracy, precision and sensitivity. The assay was validated for human plasma, mouse plasma and Dulbeccos Modified Eagles Medium (DMEM) containing varying concentrations of Foetal Bovine Serum (FBS). Matrix effects are a well-documented source of concern for chromatographic analysis, with the potential to impact various stages of the analytical process, including suppression or enhancement of ionisation. Therefore, a robustly validated LC-MS/MS analytical method is presented capable of quantifying tizoxanide in multiple matrices with minimal impact of matrix effects. The validated assay presented here was linear from 15.6ng/mL to 1000ng/mL. Accuracy and precision ranged between 102.2% and 113.5%, 100.1% and 105.4%, respectively. The presented assay here has applications in both pre-clinical and clinical research and may be used to facilitate further investigations into the application of nitazoxanide against SARS-CoV-2.