Abstract
Favipiravir is a broad-spectrum inhibitor of viral RNA polymerase. It is currently used as a possible treatment for coronavirus disease 2019 (COVID-19). Pre-clinical or clinical trials of favipiravir require robust, sensitive, and accurate bioanalytical methods for quantitation of favipiravir levels. Recently, several studies have been reported about developing a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for measuring favipiravir levels. However, these methods were validated predominantly for plasma samples, electrospray ionization was operated only in negative or positive mode, and clinical application of these methods has not been applied for patients with COVID-19. This study aimed was to develop a validated LC-MS/MS method for the measurement of favipiravir levels in positive and negative electrospray ionization mode and to perform a pilot study in patients with COVID-19 receiving favipiravir to demonstrate the applicability of this method in biological samples. Simple protein precipitation was used for the extraction of favipiravir from the desired matrix. Favipiravir levels were quantitated using MS / MS with an electrospray ionization source in positive and negative multiple reaction monitoring (MRM) mode. The chromatographic detection was performed on a reverse-phase Phenomenex C18 column (50 mm × 4.6 mm, 5 µm, 100 Å) with gradient elution using 0.1% formic acid in water and 0.1% formic acid in methanol as mobile phase. The method was linear over the concentration ranges of 0.048-50 µg/mL (in negative ionization mode) and 0.062-50 µg/mL (in positive ionization mode) with a correlation coefficient (r 2 ) better than 0.998. The total run time was 3.5 min. The intra-assay and inter-assay %CV values were less than 7.2% and 8.0%, respectively. A simple, rapid and robust LC-MS / MS method was developed for the measurement of favipiravir and validation studies were performed. The validated method was successfully applied for drug level measurement in COVID-19 patients receiving favipiravir.
Keywords: COVID-19; Favipiravir; Tandem mass spectrometry; Validation.
【저자키워드】 COVID-19, Favipiravir, validation, Tandem mass spectrometry, 【초록키워드】 Treatment, coronavirus disease, Coronavirus disease 2019, mass spectrometry, clinical trial, Favipiravir, validation, Viral RNA, inhibitor, LC-MS, Tandem mass spectrometry, Concentration, COVID-19 patient, biological samples, water, Formic acid, methanol, pilot study, plasma samples, extraction, validation study, protein precipitation, applicability, quantitation, positive, polymerase, correlation coefficient, simple, tandem, electrospray ionization, robust, performed, was used, develop, reported, receiving, linear, applied, was performed, less, drug level, patients with COVID-19, Phenomenex, 【제목키워드】 Favipiravir, development, human serum, LC-MS,