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Comparing the present method to other reported assays
Recently, Alvarez et al. developed an HPLC-MS/MS method and applied it to assess a patient who was intravenously administered remdesivir. However, the re- quired plasma volume was 50 μL and matrix effects were not reported. Moreover, detailed recovery, uncertainty of measurement, and incurred sample reanalysis were not determined. A comparison of the method reveals several advantages of the present HPLC-MS/MS method, such as simple plasma preparation, satisfactory matrix effect and recovery, detailed pharmacokinetic application, and ro- bustness owing to measuring the uncertainty and incurred sample reanalysis (Table 5).
Conclusions A high-throughput, robust, sensitive, and reproducible HPLC-MS/MS method was developed and fully validated for the quantitation of Nuc, the active metabolite of remdesivir, in plasma. Worsening of the COVID-19 pan- demic has necessitated the development of an HPLC-MS/ MS method that can accurately quantitate Nuc to provide plasma exposure information, because obtaining this informa- tion has become a bottleneck in the clinical treatment of COVID-19. The optimized HPLC-MS/MS method reported here offers significant advantages including cost-effective, high-throughput sample preparation (protein precipitation ex- traction), and requiring only small volume of biological ma- trix (20 μL). Furthermore, this method meets the requirements for extraction recovery and matrix effects. This quantitative method was also successfully applied in a rat pharmacokinet- ics study. This reliable method will provide sufficient refer- ence and mirror for pharmacokinetics and clinical efficacy in patients with COVID-19


Original text

Comparing the present method to other reported assays
Recently, Alvarez et al. developed an HPLC-MS/MS method and applied it to assess a patient who was intravenously administered remdesivir. However, the re- quired plasma volume was 50 μL and matrix effects were not reported. Moreover, detailed recovery, uncertainty of measurement, and incurred sample reanalysis were not determined. A comparison of the method reveals several advantages of the present HPLC-MS/MS method, such as simple plasma preparation, satisfactory matrix effect and recovery, detailed pharmacokinetic application, and ro- bustness owing to measuring the uncertainty and incurred sample reanalysis (Table 5).
Conclusions A high-throughput, robust, sensitive, and reproducible HPLC-MS/MS method was developed and fully validated for the quantitation of Nuc, the active metabolite of remdesivir, in plasma. Worsening of the COVID-19 pan- demic has necessitated the development of an HPLC-MS/ MS method that can accurately quantitate Nuc to provide plasma exposure information, because obtaining this informa- tion has become a bottleneck in the clinical treatment of COVID-19. The optimized HPLC-MS/MS method reported here offers significant advantages including cost-effective, high-throughput sample preparation (protein precipitation ex- traction), and requiring only small volume of biological ma- trix (20 μL). Furthermore, this method meets the requirements for extraction recovery and matrix effects. This quantitative method was also successfully applied in a rat pharmacokinet- ics study. This reliable method will provide sufficient refer- ence and mirror for pharmacokinetics and clinical efficacy in patients with COVID-19


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