Simultaneous quantification of seven repurposed COVID-19 drugs remdesivir (plus metabolite GS-441524), chloroquine, hydroxychloroquine, lopinavir, ritonavir, favipiravir and azithromycin by a two-dimensional isotope dilution LC-MS/MS method in human serum.


Journal

Journal of pharmaceutical and biomedical analysis
ISSN: 1873-264X
Titre abrégé: J Pharm Biomed Anal
Pays: England
ID NLM: 8309336

Informations de publication

Date de publication:
20 Mar 2021
Historique:
received: 09 10 2020
revised: 30 12 2020
accepted: 24 01 2021
pubmed: 7 2 2021
medline: 27 2 2021
entrez: 6 2 2021
Statut: ppublish

Résumé

The present COVID-19 pandemic has prompted worldwide repurposing of drugs. The aim of the present work was to develop and validate a two-dimensional isotope-dilution liquid chromatrography tandem mass spectrometry (ID-LC-MS/MS) method for accurate quantification of remdesivir and its active metabolite GS-441524, chloroquine, hydroxychloroquine, lopinavir, ritonavir, favipiravir and azithromycin in serum; drugs that have gained attention for repurposing in the treatment of COVID-19. Following protein precipitation, samples were separated with a two-dimensional ultra-high performance liquid chromatography (2D-UHPLC) setup, consisting of an online solid phase extraction (SPE) coupled to an analytical column. For quantification, stable isotope-labelled analogues were used as internal standards for all analytes. The method was validated on the basis of the European Medicines Agency bioanalytical method validation protocol. Detuning of lopinavir and ritonavir allowed simultaneous quantification of all analytes with different concentration ranges and sensitivity with a uniform injection volume of 5 μL. The method provided robust validation results with inaccuracy and imprecision values of ≤ 9.59 % and ≤ 11.1 % for all quality controls. The presented method is suitable for accurate and simultaneous quantification of remdesivir, its metabolite GS-441525, chloroquine, hydroxychloroquine, lopinavir, ritonavir, favipiravir and azithromycin in human serum. The quantitative assay may be an efficient tool for the therapeutic drug monitoring of these potential drug candidates in COVID-19 patients in order to increase treatment efficacy and safety.

Sections du résumé

BACKGROUND BACKGROUND
The present COVID-19 pandemic has prompted worldwide repurposing of drugs. The aim of the present work was to develop and validate a two-dimensional isotope-dilution liquid chromatrography tandem mass spectrometry (ID-LC-MS/MS) method for accurate quantification of remdesivir and its active metabolite GS-441524, chloroquine, hydroxychloroquine, lopinavir, ritonavir, favipiravir and azithromycin in serum; drugs that have gained attention for repurposing in the treatment of COVID-19.
METHODS METHODS
Following protein precipitation, samples were separated with a two-dimensional ultra-high performance liquid chromatography (2D-UHPLC) setup, consisting of an online solid phase extraction (SPE) coupled to an analytical column. For quantification, stable isotope-labelled analogues were used as internal standards for all analytes. The method was validated on the basis of the European Medicines Agency bioanalytical method validation protocol.
RESULTS RESULTS
Detuning of lopinavir and ritonavir allowed simultaneous quantification of all analytes with different concentration ranges and sensitivity with a uniform injection volume of 5 μL. The method provided robust validation results with inaccuracy and imprecision values of ≤ 9.59 % and ≤ 11.1 % for all quality controls.
CONCLUSION CONCLUSIONS
The presented method is suitable for accurate and simultaneous quantification of remdesivir, its metabolite GS-441525, chloroquine, hydroxychloroquine, lopinavir, ritonavir, favipiravir and azithromycin in human serum. The quantitative assay may be an efficient tool for the therapeutic drug monitoring of these potential drug candidates in COVID-19 patients in order to increase treatment efficacy and safety.

Identifiants

pubmed: 33548872
pii: S0731-7085(21)00047-9
doi: 10.1016/j.jpba.2021.113935
pmc: PMC7843035
pii:
doi:

Substances chimiques

Amides 0
Antiviral Agents 0
Furans 0
Isotopes 0
Pyrazines 0
Pyrroles 0
Triazines 0
GS-441524 1BQK176DT6
Lopinavir 2494G1JF75
remdesivir 3QKI37EEHE
Adenosine Monophosphate 415SHH325A
Hydroxychloroquine 4QWG6N8QKH
Azithromycin 83905-01-5
Chloroquine 886U3H6UFF
favipiravir EW5GL2X7E0
Adenosine K72T3FS567
Ritonavir O3J8G9O825
Alanine OF5P57N2ZX

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

113935

Informations de copyright

Copyright © 2021 Elsevier B.V. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of Competing Interest The authors have no conflict of interest to declare.

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Auteurs

Katharina Habler (K)

Institute of Laboratory Medicine, University Hospital, LMU Munich, Germany.

Mathias Brügel (M)

Institute of Laboratory Medicine, University Hospital, LMU Munich, Germany.

Daniel Teupser (D)

Institute of Laboratory Medicine, University Hospital, LMU Munich, Germany.

Uwe Liebchen (U)

Department of Anesthesiology, University Hospital, LMU Munich, Germany.

Christina Scharf (C)

Department of Anesthesiology, University Hospital, LMU Munich, Germany.

Ulf Schönermarck (U)

Department of Medicine IV, University Hospital, LMU Munich, Germany.

Michael Vogeser (M)

Institute of Laboratory Medicine, University Hospital, LMU Munich, Germany.

Michael Paal (M)

Institute of Laboratory Medicine, University Hospital, LMU Munich, Germany. Electronic address: Michael.Paal@med.uni-muenchen.de.

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Classifications MeSH