Development and validation of simple HPTLC method for the determination of new hepatitis C subtype 4 antiviral agents in their tablet dosage form

A new, simple, precise, accurate and selective high-performance thin layer chromatographic (HPTLC) method has been developed and validated for the simultaneous determination of ledipasvir and sofosbuvir in tablet dosage form. Chromatographic separation was carried out on Merck TLC aluminum sheets of silica gel 60F254 using Ethyl acetate: Hexane: Methanol in the ratio of 8: 1.25: 0.75 (% v/v/v) as mobile phase followed by densitometric measurement at 256 nm.The method was validated in terms of linearity, accuracy, precision, limit of detection, limit of quantification and specificity in accordance with International Conference on Harmonization (ICH) guidelines. The calibration curve was found to be linear between60 to 1980 and 45 to 3600 ng/band for ledipasvir and sofosbuvir, respectively with significantly high value of regression coefficient (r2 > 0.9999) with linear and homoscedastic residuals. The limits of detection and quantitation were found to be 16.5 and 50 ng/band, respectively for ledipasvir and 13 and 39.5 ng/band, respectively for sofosbuvir. Comparative study was performed between the developed HPTLC method and the reported HPLC method. The quantitative results of both analytical methods did not show statistically significant difference, whereas the developed HPTLC method is both time and cost effective.