Development and validation of stability indicating HPLC method for favipiravir used in the treatment of the Covid-19 disease

Year 2024, Volume: 54 Issue: 2, 223 - 232, 26.08.2024

Büşra Gülşen , Sıdıka Ertürk Toker

https://doi.org/10.26650/IstanbulJPharm.2024.1368223

Abstract

Background and Aims: Favipiravir (FAV) is one of the active pharmaceutical ingredients used in the treatment of patients suffering from Covid-19. The epidemic started in 2019 and is still continuing all over the world. In this study, an analysis method was developed and validated for the simultaneous analysis of FAV and its degradation impurities.

Methods: The stationary phase of the developed method was determined using Kinetex® EVO C18 column and the mobile phase was pH3.0phosphate buffer:acetonitrile (90:10; v/v). Chromatographic separations were carried out at 30 °C column temperature and samples were monitored by a UV-Visible detector with a wavelength of 222 nm at 0.5 mL/min flow rate.

Results: Total analysis time was 25 minutes; FAV retention time was approximately 9 minutes. The retention times of major impurities formed under alkaline, acidic, oxidative conditions were observed at about 4, 5, 7 and,12 minutes (RRT 0.51, 0.54, 0.76, 1.31), respectively. According to the validation data, the linearity range was obtained as 0.030– 0.750 μg/mL, the limit of quantitation and the limit of detection were 0.030 μg/mL and 0.010 μg/mL, respectively. Percentage relative standard deviation values obtained in intra-day and between day repeatability studies were determined as 0.17% and 0.28%, respectively, and the average recovery value was found to be 99.46%.

Conclusion: This validated method has been successfully applied to the determination of all possible degradation impurities of FAV that increase under stress conditions such as high temperature, humidity and photodegradation from tablet form. The developed HPLC method is extremely suitable for the routine analysis of this drug used in the treatment of the Covid-19 disease, especially in terms of speed and convenience.

Keywords

Covid-19, favipiravir, stability indicating method, HPLC, validation

References

• Ali, S.N.S., Mobina, L., Mehfuza, M., Seema, P., Ahmed, A., & Khan, G.J. (2021). Analytical method development and validation and forced degradation stability-indicating studies of favipiravir by RP-HPLC and UV in bulk and pharmaceutical dosage form. Jour-nal of Pharmaceutical Research International, 33, 254-271. doi: 10.9734/JPRI/2021/v33i48B33283 google scholar
• Baranovich, T., Wong, S. S., Armstrong, J., Marjuki, H., Webby, R. J., Webster, R. G., & Govorkova, E. A. (2013). T-705 (favipiravir) induces lethal mutagenesis in influenza A H1N1 viruses in vitro. Journal of Virology, 87(7), 3741-3751. doi:http://dx.doi.org/10.1128/JVI.02346-12T google scholar
• Bulduk, İ. (2020). HPLC-UV method for quantification of Favipiravir in pharmaceutical formulations. Acta Chromatographica, 33, 209215. doi:10.1556/1326.2020.00828 google scholar
• Cuiyan, L., Yuanyuan, Z., Lichao, B., Yangjin, L., & Lei, L. (2015). google scholar
• Content determination of favipiravir tablets by HPLC. China Pharmacist, 7, 1231-1233. google scholar
• Delong, L., Abdelnabi, R., & Neyts, J. (2018). Favipiravir as a potential countermeasure against neglected and emerging RNA viruses. An-tiviral Research, 153, 85-94. doi:10.1016/j.antiviral.2018.03.003 google scholar
• Dolan, J. (2011) (Accessed 2 March 2021) A Guide to HPLC and LC-MS Buffer Selection, https://www.hplc.eu/Downloads/ACE_Guide_BufferSelection.pdf. google scholar
• Drugbank, Favipiravir (2021, March 1), https://go.drugbank.com/drugs/DB12466 google scholar
• Guangling, F., Wenjuan, D., Yuxiao, D., Ren-Yang, Z., Yan, G., Chong-Gang, D., & Jinrui, S. (2015). HPLC method for mea-suring related substances in Favipiravir (Chinese Patent No CN104914185A). Shandong Academy of Pharmaceutical Sci-ences, https://patents.google.com/patent/CN104914185A/en google scholar
• Guangling, F., Wenjuan, D., Yuxiao, D., Ren-Yang, Z., Yan, G., Chong-Gang, D., & Jinrui, S. (2016). A kind of Favipiravir has the HPLC assay method of related substance (Chinese Patent No. CN104914185B) Shandong Academy of Pharmaceutical Sci-ences, https://patents.google.com/patent/CN104914185B/en google scholar
• Guidance for Industry ANDAs: Impurities in Drug Prod-ucts, Food and Drug Administration, (2010) (Accessed 23 March 2021) https://www.fda.gov/files/drugs/published/ANDAs-Impurities-in-Drug-Products.pdf google scholar
• ICH Q2 (R1):Validation of Analytical Procedures: Text and Method-ology, 2005, European Medicines Agency, (Accessed 12 March 2021) https://www.ema.europa.eu/en/documents/scientific-guideline/ich-q-2-r1-validation-analytical-procedures-text-methodology-step-5en.pdf google scholar
• Kiso, M., Takahashi, K., Sakai-Tagawa, Y., Shinya, K., Sakabe, S., Le, M., Ozawa, M., Furuta, Y., Kawaoka, Y. (2010). T-705 (Favipi-ravir) Activity against Lethal H5N1 Influenza A Viruses. Pro-ceedings of the National Academy of Sciences of the United States of America, 107, 882-887. doi: 10.1073/pnas.0909603107 google scholar
• Megahed, S. M., Habib, A. A., Hammad, S. F., & Kamal, A. H. (2020). Experimental design approach for development of spectrofluorimetric method for determination of favipiravir; a potential therapeutic agent against COVID-19 virus: Applica-tion to spiked human plasma. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 249, 1386-1425. doi:10.1016/j.saa.2020.119241 google scholar
• Japan Pharmaceuticals and Medical Devices Agency, 2014 Report on the deliberation results, Avigan 200 mg Tablet, (Accessed 31 December 2020) https://www.pmda.go.jp/files/000210319.pdf google scholar
• Turkish Republic Ministry of Health, Covid-19 Informa-tion Platform, What is Covid-19? (December 2020) https://covid19.saglik.gov.tr/TR-66300/covid-19-nedir-.html google scholar
• Yegorova, A. V., Scrypynets, Y. V., Leonenko, I. I., Umetskaya, I. V., & Voitiuk, O. D. (2020). Determination of favipiravir residues on the surfaces of pharmaceutical manufacturing equipment by HPLC, luminescence and spectrophotometry. Odesa National University Herald Chemistry, 25, 45-55. doi:10.18524/2304-0947.2020.4(76).216925 google scholar
• Watanabe, T., Kiso, M., Fukuyama, S., Nakajima, N., Imai, M., Yamada, S., . . . Kawaoka, Y. (2013). Characterization of H7N9 influenza A viruses isolated from humans. Nature, 501, 551-555. doi:10.1038/nature12392 google scholar