Abstract :

The optimization of chromatographic conditions for the analysis of Ledipasvir and Sofosbuvir was successfully achieved using a Hibar C18(250×4.6mm)5µm column using 1% O-Phosphoric acid (pH 6.4): Acetonitrile (30:70 v/v) at a flow rate of 0.8 ml/min, ensuring excellent peak shape and resolution. System suitability tests confirmed compliance with essential parameters, including resolution, tailing factor, and theoretical plate count, ensuring reliability. The retention time of ledipasvir and sofosbuvir were found to be 7.1min and 3.7 min, respectively. During force degradation, drug product was exposed to hydrolysis (acid and base hydrolysis), oxidation, thermal degradation and photo degradation. Both the drugs were not degraded under thermal, oxidative, photolytic, acid and neutral hydrolytic conditions, but Sofosbuvir showed degradation under alkaine hydrolytic condition with a retention time 2.8 min and 4.1 min, respectively. The degraded products of Ledipasvir and Sofosbuvir were well resolved from the individual bulk drug response. The specificity of the method was confirmed by peak purity profile of the resolved peaks. In conclusion, the developed chromatographic method for Ledipasvir and Sofosbuvir is precise, accurate, linear, sensitive, and robust, making it suitable for routine pharmaceutical analysis and quality control. The validation results confirm compliance with regulatory standards, ensuring reliable identification and quantification of these antiviral drugs. Given its high stability and reproducibility, this method provides an efficient and dependable approach for analyzing Ledipasvir and Sofosbuvir in pharmaceutical formulations.

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Keywords :

Accuracy, Chromatographic Optimization, Degradation, Ledipasvir, Linearity, Pharmaceutical Analysis, Precision, Retention Time, Robustness, Sofosbuvir.

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References :

1. Beckett AH, Stenlake JB. Practical pharmaceutical chemistry. 4^th Vol ΙΙ. New Delhi: CBS publishers and distributors; 2005.p.1-3,300301.
2. Napoleon AA, pharmaceutical titrimetric analysis. 1^st Kalaimani publisers and distributers; 2006. P. 1-2.
3. Sethi PD. HPTLC: High performance thin layer Chromatography. Vol Ι. Quantitative Analysis of Pharmaceutical formulations. New Delhi: CBS publishers and distributers: 1997.p. 2-81.
4. Snyder LR, Kirkland JJ, Glajch JL. Practical HPLC method development. 2^nd New York: John Wiley& Sons, Inc; 1997. p.1-15, 205-13, 234-66.
5. Satinder A, Michael WD. Hand book of Pharmaceutical Analysis by HPLC. 6^th New Delhi: Elseveier; 2009. P. 22-39.
6. The United States Pharmacopoeia, 27^th Rockville: United state Pharmacopoeial Convention; 2004. P. 221-82
7. ICH harmonized tripartite guidelines. Stability testing of new drugs substance and products Q2( R1) 2005 Nov. Geneva, Switzerland.
8. Bakshi M, Singh S, Development of validated stability-indicating assay methodscriticalreview.J. Pharm Biomed Anal.2002 Jun 15;28(6): 1011-40.
9. ICH Q2B Validation of Analytical Procedure Methodology. International Conference on Harmonization. Geneva: 1996
10. Mohan Vikas P, Satyanarayana T, Vinod Kumar D, Mounika E, Sri Latha M. Development and validation of new rp-hplc method for the determination of of Sofosbuvir in pure form, World J Pharm Pharm Sci. 2016; 5: 775-81
11. Devila J, Durgaprasad B, Narottam Pal, Srinivasa Rao A. New method development and validation for the determination of Ledipasvir in bulk drug form by using reverse phase hplc technique, World J Pharm Pharm Sci. 2016; 8: 1312-21.
12. Chenwei Pana, Yongping Chenb, Weilai Chenc, Guangyao Zhoua, ingxiang Jina. Simultaneous determination of ledipasvir, Sofosbuvir and its metabolite in rat plasma by UPLC–MS/MS and its application to a pharmacokinetic study, J. Chromatogr. B. 2016; 1009: 255-59.
13. Nebsen M, Elzanfaly S. Stability-Indicating Method and LC–MS-MS Characterization of Forced Degradation Products of Sofosbuvir, J CHROMATOGR SCI. 2016; 1-10.
14. Ehab Elkady F, Ahmed Aboelwafa A. A Rapid and Optimized LC-MS/MS Method for the Simultaneous Extraction and Determination of Sofosbuvir and Ledipasvir in Human Plasma, J. AOAC Int. 2016; 99: 1252-59.
15. Debasish Swain, Gananadhamu Samanthula. Study on the forced degradation behaviour of ledipasvir: Identification of major degradation products using LC-QTOF-MS/MS and NMR, J Pharm Biomed Anal. 2017