Abstract
A novel, fast and sensitive LC-MS/MS method was developed and validated for the bioanalysis of the antiviral agent favipiravir (FAV); a promising candidate for treatment of SARS-CoV-2 (COVID-19) in human plasma using pyrazinamide as an internal standard (IS). Simple protein precipitation was adopted for plasma sample preparation using methanol. Chromatographic separation was accomplished on Eclipse plus C 18 column (50 × 4.6 mm, 3.5 μm) using a mobile phase composed of methanol-0.2 % acetic acid (20:80, v/v) pumped at a flow rate 0.6 mL/min in an isocratic elution mode. The API4500 triple quadrupole tandem mass spectrometer was operated with multiple-reaction monitoring (MRM) in negative electrospray ionization interface for FAV and positive for IS. The MRM function was used for quantification, with the transitions set at m/z 156.00→ 113.00 and m/z 124.80→ 81.00 for FAV and IS. The method was optimized and fully validated in accordance to US-FDA guidelines. Linearity was acquired over a concentration range of 100.0-20000.0 ng/mL by computing using weighted linear regression strategy (1/x 2 ). The proposed method was effectively applied for the pharmacokinetic evaluation of FAV and to demonstrate the bioequivalence of a new FAV formulation (test) and reference product in healthy Egyptian human volunteers.
Keywords: Bioequivalence study; COVID-19; Favipiravir; Human plasma; LC–MS/MS; SARS-CoV-2.
【저자키워드】 COVID-19, SARS-CoV-2, Favipiravir, Bioequivalence study, Human plasma, LC–MS/MS, 【초록키워드】 Treatment, SARS-CoV-2, Antiviral, Human, Favipiravir, bioanalysis, Linear regression, antiviral agent, quantification, volunteers, pharmacokinetic, Bioequivalence, Human plasma, Concentration, linearity, flow rate, methanol, mass spectrometer, acetic acid, pyrazinamide, protein precipitation, positive, simple, plasma sample, electrospray ionization, US-FDA, was used, healthy, composed, applied, adopted, acetic, Eclipse, isocratic, m/z, pumped, 【제목키워드】 Treatment, Antiviral, Favipiravir, SARS-CoV-2 virus, application, Volunteer,