The development and validation of a novel LC-MS/MS method for the simultaneous quantification of Molnupiravir and its metabolite ß-d-N4-hydroxycytidine in human plasma and saliva.
The development and validation of a novel LC-MS/MS method for the simultaneous quantification of Molnupiravir and its metabolite ß-d-N4-hydroxycytidine in human plasma and saliva.
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DOI：
10.1016/j.jpba.2021.114356
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发表时间：
2021-11-30
影响因子：
3.4
通讯作者：
Khoo S
中科院分区：
文献类型：
--
作者：
Amara A;Penchala SD;Else L;Hale C;FitzGerald R;Walker L;Lyons R;Fletcher T;Khoo S
鉴于最近的全球大流行，为治疗无并发症的流感患者而开发的 Molnupiravir (MPV) 或 EIDD-2801 目前正在试验用于治疗由高致病性冠状病毒（包括 COVID-19）引起的感染。开发并验证了一种灵敏的 LC-MS/MS 方法，用于同时定量人血浆和唾液中的 MPV 及其代谢物 ß-d-N4-羟基胞苷 (NHC)。使用乙腈通过蛋白质沉淀从基质中提取分析物。随后进行干燥并随后将重构的溶液注射到柱上。使用极性 Atlantis C18 柱，采用 1 mM 醋酸铵水溶液 (pH4.3) 和 1 mM 醋酸铵乙腈溶液进行梯度洗脱，实现色谱分离。使用 SRM 在负电离模式下进行分析物检测。使用稳定同位素标记 (SIL) 内标 (IS) 进行分析。 m/z 转换为：MPV (328.1→126.0)、NHC (258.0→125.9) 和 MPV-SIL (331.0→129.0)、NHC-SIL (260.9→128.9)。血浆和唾液的验证均在 2.5–5000 ng/ml 的线性范围内。四种不同浓度的精密度和准确度（日内和日间）均为 15%；血浆和唾液中两种分析物的回收率分别为 95% 至 100% 和 65-86%。临床药代动力学研究正在进行中，利用该方法测定 COVID-19 感染患者的 MPV 及其代谢物。
In light of the recent global pandemic, Molnupiravir (MPV) or EIDD-2801, developed for the treatment of patients with uncomplicated influenza, is now being trialled for the treatment of infections caused by highly pathogenic coronaviruses, including COVID-19. A sensitive LC-MS/MS method was developed and validated for the simultaneous quantification of MPV and its metabolite ß-d-N4-hydroxycytidine (NHC) in human plasma and saliva. The analytes were extracted from the matrices by protein precipitation using acetonitrile. This was followed by drying and subsequently injecting the reconstituted solutions onto the column. Chromatographic separation was achieved using a polar Atlantis C18 column with gradient elution of 1 mM Ammonium acetate in water (pH4.3) and 1 mM Ammonium acetate in acetonitrile. Analyte detection was conducted in negative ionisation mode using SRM. Analysis was performed using stable isotopically labelled (SIL) internal standards (IS). The m/z transitions were: MPV (328.1→126.0), NHC (258.0→125.9) and MPV-SIL (331.0→129.0), NHC-SIL (260.9→128.9). Validation was over a linear range of 2.5–5000 ng/ml for both plasma and saliva. Across four different concentrations, precision and accuracy (intra- and inter-day) were 15%; and recovery of both analytes from plasma and saliva was between 95% and 100% and 65–86% respectively. Clinical pharmacokinetic studies are underway utilising this method for determination of MPV and its metabolite in patients with COVID-19 infection.
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影响因子：
7.6
作者：
Painter, George R.;Bowen, Richard A.;Kolykhalov, Alexander A.
通讯作者：
Kolykhalov, Alexander A.
DOI：
10.1016/j.jchromb.2009.09.014
发表时间：
2009-11-01
影响因子：
3
作者：
Perogamvros, Ilias;Owen, Laura J.;Keevil, Brian G.
通讯作者：
Keevil, Brian G.
DOI：
10.1016/s1473-3099(14)70717-0
发表时间：
2014-07
期刊：
The Lancet. Infectious diseases
影响因子：
--
作者：
Haffizulla J;Hartman A;Hoppers M;Resnick H;Samudrala S;Ginocchio C;Bardin M;Rossignol JF;US Nitazoxanide Influenza Clinical Study Group
通讯作者：
US Nitazoxanide Influenza Clinical Study Group
影响因子：
158.5
作者：
Beigel, John H.;Tomashek, Kay M.;Lane, H. Clifford
通讯作者：
Lane, H. Clifford
影响因子：
2.2
作者：
Israelsson, Marlen;Brattsand, Ralph;Brattsand, Goran
通讯作者：
Brattsand, Goran