Abstract
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: 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: 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: 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.
Keywords: Antiviral therapy; Isotope dilution liquid chromatography tandem mass spectrometry (ID-LC–MS/MS); Therapeutic drug monitoring.
【저자키워드】 antiviral therapy, Isotope dilution liquid chromatography tandem mass spectrometry (ID-LC–MS/MS), Therapeutic Drug Monitoring, 【초록키워드】 COVID-19, Treatment, antiviral therapy, mass spectrometry, protocol, Chloroquine, Azithromycin, Hydroxychloroquine, COVID-19 pandemic, Lopinavir, Ritonavir, drugs, Remdesivir, Favipiravir, drug, European Medicines Agency, serum, sensitivity, Liquid chromatography, GS-441524, Efficacy and safety, quantification, human serum, metabolite, Tandem mass spectrometry, Concentration, COVID-19 patient, Dilution, solid phase, Therapeutic Drug Monitoring, Volume, two-dimensional, injection, drug candidate, protein precipitation, LOPINAVIR AND RITONAVIR, quantitative assay, inaccuracy, analogue, Bioanalytical Method Validation, controls, robust, analytical column, develop, provided, were used, separated, Isotope, therapeutic drug, 【제목키워드】 COVID-19, drug, quantification, metabolite, Dilution, two-dimensional, Simultaneous, Seven,