Development and validation of stability indicating RP-HPLC method for the simultaneous estimation of Sofosbuvir and Ledipasvir in bulk and their combined dosage form

Abstract

Objective: A simple, specific, accurate and economic reverse phase liquid chromatographic method was developed for the simultaneous estimation of Sofosbuvir and Ledipasvir in bulk and tablet dosage form. Method: The method has shown adequate separation of Sofosbuvir and Ledipasvir from their degradation products. Separation was achieved on a Luna C18, 250 mm × 4.6 mm, 5 μm Column at wavelength of 227 nm, using a mobile phase acetonitrile: Triethylamine Buffer (pH-2.5) (50:50) in an isocratic elution mode at a flow rate of 1.0 ml/min. Results: The retention time for Sofosbuvir and Ledipasvir is found to be 4.905 and 2.751 min correspondingly. The above drug combination is subjected to acidic, base, neutral hydrolysis, thermal and photolytic stress environment. Thus stressed samples are analyzed by the proposed analytical method. Quantitation was achieve with UV detection at 227 nm based on peak area with linear calibration curve at concentration range 1–15 μg/ml for Sofosbuvir and 0.25–3.75 μg/ml for Ledipasvir. The LOD's were 0.25 and 0.0625 for Sofosbuvir and Ledipasvir respectively. The LOQ's were found to be 0.5 for Sofosbuvir and 0.505 for Ledipasvir. Conclusion: The proposed method was established to be precise and stability indicating as no interfering peaks of degrades and excipient was observed. The proposed method is therefore suitable for purpose in quality-control laboratories for quantitative analysis of both the drugs individually and in combined dosage form, as it is simple and rapid with tremendous precision and accuracy. Keywords: Sofosbuvir, Ledipasvir, RP-HPLC, Stability indicating