As of 2020, the development of both pharmaceuticals and vaccines remains urgent to overcome the global coronavirus disease 2019 (COVID-19) pandemic. Favipiravir (brand name: Avigan®), a promising drug candidate for COVID-19, is classified as an antiinfluenza drug, evaluated and developed for both novel and re-emerging influenza viruses. (1), (2) Favipiravir undergoes renal excretion, eliminated in the urine mainly as a hydroxide. Notably, the plasma levels of this drug are difficult to control owing to its once daily dosing regimen. (3) Consequently, the accurate monitoring of drug levels is crucial.
In this report, we introduce a unique approach for the quantitative high sensitivity plasma analysis of favipiravir using only a standard high-pressure liquid chromatography (HPLC) setup without mass spectrometry.
HPLC Analysis
Favipiravir *1 was purchased from Alsachim. The calibration curve and quality control (QC) samples were prepared by spiking healthy human plasma with favipiravir. The measurement was performed using HPLC analytical conditions shown in Table 1. The time program of gradient elution is shown in Table 2. Favipiravir was separated using an HPLC instrument fitted with Shim-pack Scepter™ C18-120 with a guard column. The chromatograms are shown in Fig. 3. A calibration curve was generated using favipiravir standard solutions of at 1, 10, 25, 50, and 100 μg/mL (n = 6) spiked in plasma.
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