Development and Validation of a HILIC–HPLC Method for the Quantitative Determination of Favipiravir in Tablet Formulations

Abstract

Objective: A validated Hydrophilic Interaction Liquid Chromatography (HILIC) method was developed for the quantitative determination of Favipiravir. Methods: The chromatographic analysis was performed using a high-performance liquid chromatography (HPLC) system comprising a diode array detector, vacuum pump, column thermostat, and autosampler (Agilent 1260, Agilent Technologies, Santa Clara, CA, USA). The analysis was conducted on a ZORBAX RRHD HILIC Plus analytical column (95 Å, 2.1 × 150 mm, 1.8 µm). The mobile phase consisted of acetonitrile and water (80:20, v/v). Chromatographic separation was achieved under isocratic elution at a flow rate of 0.8 mL/min, and Favipiravir was detected at a wavelength of 322 nm. Results: The retention time of Favipiravir was 1.8317 ± 0.002 min, with a limit of detection (LOD) of 0.14 μg/mL and a limit of quantification (LOQ) of 0.47 μg/mL. The regression equation was y = 107.54x + 17.034 with a correlation coefficient (r²) of 0.9997. Conclusion: The proposed method was validated in terms of specificity, linearity, accuracy, precision, LOD, and LOQ, and was successfully applied for the quantitative analysis of Favipiravir in tablet formulations.

Keywords

• Favipiravir
• HILIC
• HPLC

Tablet Formülasyonlarında Favipiravir’in Kantitatif Belirlenmesi İçin HILIC–HPLC Yönteminin Geliştirilmesi ve Validasyonu

Abstract

Amaç: Favipiravir’in kantitatif belirlenmesi için valide edilmiş bir Hidrofilik Etkileşimli Sıvı Kromatografi (HILIC) yöntemi geliştirilmiştir. Yöntemler: Kromatografik analiz, diyot dizi dedektörü, vakum pompası, kolon termostatı ve otomatik örnekleyici içeren bir yüksek performanslı sıvı kromatografi (HPLC) sistemi (Agilent 1260, Agilent Technologies, Santa Clara, CA, ABD) kullanılarak gerçekleştirilmiştir. Analiz, ZORBAX RRHD HILIC Plus analitik kolonunda (95 Å, 2.1 × 150 mm, 1.8 µm) yapılmıştır. Mobil faz %80 asetonitril ve %20 sudan (v/v) oluşmuştur. Ayrım, 0.8 mL/dk akış hızında izokratik elüsyonla sağlanmış ve Favipiravir 322 nm dalga boyunda tespit edilmiştir. Bulgular: Favipiravir’in alıkonma süresi 1.8317 ± 0.002 dakika olarak belirlenmiş; tespit limiti (LOD) 0.14 µg/mL ve miktar tayin limiti (LOQ) 0.47 µg/mL olarak hesaplanmıştır. Kalibrasyon eğrisi denklemi y = 107.54x + 17.034 olup korelasyon katsayısı (r²) 0.9997 olarak bulunmuştur. Sonuç: Önerilen yöntem; seçicilik, doğrusallık, doğruluk, kesinlik, LOD ve LOQ açısından valide edilmiş ve Favipiravir’in tablet formülasyonlarında kantitatif analizinde başarıyla uygulanmıştır.

Keywords

• Favipiravir
• HILIC
• HPLC

References

1. 1. Venkatasubbaiah M, Dwarakanadha Reddy P, Satyanarayana SV. Literature-based review of the drugs used for the treatment of COVID-19. Curr Med Pract. 2020;10(3):100-109. [CrossRef]
2. 2. Megahed SM, Habib AA, Hammad SF, Kamal AH. Experimental design approach for development of spectrofluorimetric method for determination of favipiravir; a potential therapeutic agent against COVID-19 virus: Application to spiked human plasma. Spectrochim Acta A. 2021;249:119241. [CrossRef]
3. 3. Mikhail IE, Elmansi H, Belal F, Ibrahim AE. Green micellar solvent-free HPLC and spectrofluorimetric determination of favipiravir as one of COVID-19 antiviral regimens. Microchem J. 2021;165:106189. [CrossRef]
4. 4. Abdallah IA, Hammad SF, Bedair A, Mansour FR. Menthol-assisted homogenous liquid-liquid microextraction for HPLC/UV determination of favipiravir as an antiviral for COVID-19 in human plasma. J Chromatogr B. 2021;123087. CrossRef]
5. 5. Zhong Y, Jiao Z, Yu Y. Simultaneous determination of mycophenolic acid and valproic acid based on derivatization by high‐performance liquid chromatography with fluorescence detection. Biomed. Chromatogr. 2006;20(4):319-326. [CrossRef]
6. 6. McCalley DV. Understanding and manipulating the separation in hydrophilic interaction liquid chromatography. J Chromatogr A. 2017;1523:49-71. [CrossRef]
7. 7. Shirley CC. Recent progress in carbohydrate separation by high-performance liquid chromatography based on hydrophilic interaction. J Chromatogr A. 1996:720:75-91. [CrossRef] 8. Badgett JM, Boyes, B, Orlando R. The separation and quantitation of peptides with and without oxidation of methionine and deamidation of asparagine using hydrophilic interaction liquid chromatography with mass spectrometry (HILIC-MS). J Am Soc Mass Spectrom. 2017;28(5):818-826. [CrossRef] 9. Olsen BA. Hydrophilic interaction chromatography using amino and silica columns for the determination of polar pharmaceuticals and impurities. J Chromatogr A. 2001;913:113-122. [CrossRef]
8. 10. Bacalum E, Tanase M, Cheregi M, Aboul-Enein HY, David V. Retention mechanism in zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) studied for highly polar compounds under different elution conditions. Rev Roum Chim. 2016;61:531-539.

9. 11. Internatıonal Conference on Harmonısatıon of Technıcal Requırements for Regıstratıon of Pharmaceutıcals for Human Use. ICH Harmonısed Trıpartıte Guıdelıne Valıdatıon of Analytıcal Procedures: Text and Methodology Q2(R1), 2005. [CrossRef]
10. 12. Bulduk İ. HPLC-UV method for quantification of favipiravir in pharmaceutical formulations. Acta Chromatogr. 2021;33(3);209-215. [CrossRef]
11. 13. Morsy MI, Nouman EG, Abdallah YM, et al. A novel LC-MS/MS method for determination of the potential antiviral candidate favipiravir for the emergency treatment of SARS-CoV-2 virus in human plasma: Application to a bioequivalence study in Egyptian human volunteers. J Pharm Biomed Anal. 2021;199:114057. [CrossRef]
12. 14. Rezk RM, Badr KA, Abdel‐Naby NS, Ayyad MM. A novel, rapid and simple UPLC–MS/MS method for quantification of favipiravir in human plasma: Application to a bioequivalence study. Biomed Chromatogr, 2021;35(7):e5098. [CrossRef]