Validated Microwell-Based Spectrofluorimetric Method for Quantification of Ravidasvir (New Anti-Chronic Hepatitis C Virus-GT4) in Rat Plasma and Its Application to Pharmacokinetic Study

• Main Authors: Hefnawy M, Al-Shehri M, Al-Rashood S, Hammad S, Alanazi M, Alsaif N, Mohammed M, Obaidullah A, El-Gendy M
• Format: Article
• Language: English
• Published: Dove Medical Press 2020-10-01
• Series: Drug Design, Development and Therapy
• Subjects: spectrofluorimetry; ravidasvir; rat plasma; pharmacokinetic study
• Online Access: https://www.dovepress.com/validated-microwell-based-spectrofluorimetric-method-for-quantificatio-peer-reviewed-article-DDDT

Mohamed Hefnawy,1,2 Mona Al-Shehri,1 Sara Al-Rashood,1 Sherif Hammad,3,4 Mohammed Alanazi,1 Nawaf Alsaif,1 Mostafa Mohammed,1,5 Ahmad Obaidullah,1 Manal El-Gendy1 1Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia; 2Department of Analytical Chemistry, Faculty of Pharmacy, Mansoura University, Mansoura 35516, Egypt; 3Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Helwan University, Cairo 11795, Egypt; 4Basic and Applied Sciences Institute, Egypt-Japan University of Science and Technology (E-JUST), New Borg El-Arab City 21934, Alexandria, Egypt; 5National Organization for Drug Control and Research, Cairo, EgyptCorrespondence: Manal El-GendyDepartment of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, P. O. Box 2457, Riyadh 11451, Saudi ArabiaEmail [e-mail]: Ravidasvir (RAV) has been regarded as a potent new NS5A inhibitor with a magnificent safety and tolerability in the management of genotype 4 hepatitis C virus (HCV) patients. Suitable analytical techniques are needed for the measurement of RAV in different biological matrices.Methods: We have developed a fast, sensitive and economical 96-microwell-based spectrofluorimetric technique combined with one-step protein precipitation extraction strategy for the measurement of RAV in rat plasma.Results: Under the optimum conditions, the direct relationship in rat plasma was accomplished between the RAV concentrations and the fluorescence (FL) intensity in a scope of 2.5– 200 ng/mL with 0.9998 and 0.9999 for the quantification and correlation coefficients, respectively. The lower limit of detection (LLOD) was 0.840 ng/mL and this demonstrates the high sensitivity of the proposed assay. The accuracy (RE%) ranged from 95.34% to 102.29%, and the precision (RSD%) was less than 3.59%. The recovery was ranged from 93.12% to 96.26%. The stability of RAV in rat plasma was carried out and established its good stability in the range of room conventional temperature and at long-term stability (− 80°C, 30 days). The developed technique was validated as stated by the United States Food and Drug Administration (US-FDA) guidelines for bioanalytical technique verification.Conclusion: The approved technique was effectively applied for a pharmacokinetic (PK) study after single oral gavage administration of RAV at a dose of 35 mg/kg and it could be presumed that the proposed assay can be applied to clinical trials.Keywords: spectrofluorimetry, ravidasvir, rat plasma, pharmacokinetic study