Challenges and stepwise fit-for-purpose optimization for bioanalyses of remdesivir metabolites nucleotide monophosphate and triphosphate in mouse tissues using LC-MS/MS

doi:10.1016/j.jpba.2020.113806

	Challenges and stepwise fit-for-purpose optimization for bioanalyses of remdesivir metabolites nucleotide monophosphate and triphosphate in mouse tissues using LC-MS/MS
	Hu, Wenjuan 1; Chang, Lu 1; Ke, Changqiang 1; Xie, Yuanchao 1; Shen, Jingshan 1; Tan, Bo 2; Liu, Jia 1
刊名	JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
	2021-02-05
卷号	194 页码:10
关键词	Remdesivir Nucleotide monophosphate Nucleotide triphosphate Stability Recovery LC-MS/MS
ISSN号	0731-7085
DOI	10.1016/j.jpba.2020.113806
通讯作者	Tan, Bo(tbot@163.com) ; Liu, Jia(jia.liu@simm.ac.cn)
英文摘要	Remdesivir is a prodrug of the nucleotide analogue and used for COVID-19 treatment. However, the bioanalysis of the active metabolites remdesivir nucleotide triphosphate (RTP) and its precursor remdesivir nucleotide monophosphate (RMP) is very challenging. Herein, we established a novel method to separate RTP and RMP on a BioBasic AX column and quantified them by high-performance liquid chromatography-tandem mass spectrometry in positive electrospray ionization mode. Stepwise, we optimized chromatographic retention on an anion exchange column, improved stability in matrix through the addition of 5,5'-dithiobis-(2nitrobenzoic acid) and PhosSTOP EASYpack, and increased recovery by dissociation of tight protein binding with 2 % formic acid aqueous solution. The method allowed lower limit of quantification of 20 nM for RMP and 10 nM for RTP. Method validation demonstrated acceptable accuracy (93.6%-103% for RMP, 94.5%-107% for RTP) and precision (RSD < 11.9 % for RMP, RSD < 11.4 % for RTP), suggesting that it was sensitive and robust for simultaneous quantification of RMP and RTP. The method was successfully applied to analyze RMP and RTP in mouse tissues. In general, the developed method is suitable to monitor RMP and RTP, and provides a useful approach for exploring more detailed effects of remdesivir in treating diseases. (C) 2020 Elsevier B.V. All rights reserved.
资助项目	Youth Innovation Promotion Association of the Chinese Academy of Sciences[2016263] ; Shanghai Shuguang Hospital[SGXZ-201907]
WOS关键词	CLOFARABINE TRIPHOSPHATE ; MASS-SPECTROMETRY ; QUANTIFICATION ; MONO
WOS研究方向	Chemistry ; Pharmacology & Pharmacy
语种	英语
出版者	ELSEVIER
WOS记录号	WOS:000605512600050
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/295927]
专题	中国科学院上海药物研究所
通讯作者	Tan, Bo; Liu, Jia
作者单位	1.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China 2.Shanghai Univ Tradit Chinese Med, Clin Pharmacokinet Lab, Shuguang Hosp, Shanghai 201203, Peoples R China
推荐引用方式 GB/T 7714	Hu, Wenjuan,Chang, Lu,Ke, Changqiang,et al. Challenges and stepwise fit-for-purpose optimization for bioanalyses of remdesivir metabolites nucleotide monophosphate and triphosphate in mouse tissues using LC-MS/MS[J]. JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS,2021,194:10.
APA	Hu, Wenjuan.,Chang, Lu.,Ke, Changqiang.,Xie, Yuanchao.,Shen, Jingshan.,...&Liu, Jia.(2021).Challenges and stepwise fit-for-purpose optimization for bioanalyses of remdesivir metabolites nucleotide monophosphate and triphosphate in mouse tissues using LC-MS/MS.JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS,194,10.
MLA	Hu, Wenjuan,et al."Challenges and stepwise fit-for-purpose optimization for bioanalyses of remdesivir metabolites nucleotide monophosphate and triphosphate in mouse tissues using LC-MS/MS".JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 194(2021):10.