Bioanalytical LC-MS Method Development and Validation of Favipiravir

Abstract

Favipiravir (FVP) is a broad-spectrum antiviral that selectively inhibits viral RNA-dependent RNA polymerase first trialled for the treatment of influenza infection. It has been shown to be effective against a number of RNA virus families including arenaviruses, flaviviruses and enteroviruses. A liquid chromatography tandem mass spectrometry method for the quantification of FVP in rat plasma has been developed and validated. The total chromatographic runtime is 5.0 min with retention time for Favipiravir at 2.808 min and for internal standard Remdesivir is 3.999 min. respectively. The method is validated over a dynamic linear range of 2.00 – 40.00 ng/mL for Favipiravir with a correlation coefficient of R2 0.99978. The developed bioanalytical method was validated according to USFDA guidelines and all the validation parameters were within the acceptable limit. this method was found to be an accurate, precise, sensitive, and rapid method for the determination of Favipiravir in LC-MS by using Remdesivir as an internal standard. The work shows less run time while compared with other. According to the guidelines, the intra-batch and inter-batch precision (%CV) across three levels (LQC, MQC and HQC) are less than 15%. A battery of stability studies was performed on the method, and the results were found to be within the assay variability limits throughout the entire process. Under this optimised method, the pharmacokinetic study of Favipiravir in rat plasma was successful. Furthermore, a simple precipitation method for biological sample pre-treatment was developed that was simpler, more efficient, and less expensive than previously reported methods. This study prove that the method could be used for a variety of exploratory and other preclinical studies.