Earth-friendly spectrophotometric methods for simultaneous determination of ledipasvir and sofosbuvir: Application to...

Simple, rapid, sensitive, accurate, precise and earth-friendly spectrophotometric methods were developed for the simultaneous analysis of ledipasvir (LED) and sofosbuvir (SOF) without interference of both sunset yellow dye and copovidone excipients (the most probable interferents) in their combined dosage form. These proposed methods were based on measurement of LED in synthetic mixtures and combined dosage form by first derivative (¹D) spectrophotometry at 314 nm over the concentration range of 2-50 μg mL^-1 with coefficient of determination (R²) > 0.9999, mean percentage recovery of 99.98 ± 0.62. On the other hand, SOF in synthetic mixtures and combined dosage form was determined by five methods. Method I is based on the use of ¹D spectrophotometry at 274.2 nm (zero crossing point of LED). Method II involves the application of conventional dual wavelength method (DW) at the absolute difference between SOF zero order amplitudes at 261 nm (λ_max of SOF) and 364.7 nm. At these wavelengths, the absolute difference between LED zero order amplitudes was observed to equal zero. Method III depends on isosbestic point method (ISP) in which the total concentration of both drugs was measured at isosbestic point at 262.7 nm. Concentration of SOF could be obtained by subtraction of LED concentration. While, method IV depends on absorbance correction method (absorption factor method), which is based on determination of SOF concentration at 262.7 nm (λ_ISP) and LED at 333 nm (λ_max of LED). Finally, method V depends on absorbance ratio method (Q-analysis) in which 262.7 nm (λ_ISP) and 261 nm (λ_max of SOF) were selected to determine SOF concentration. The linearity range for all methods for SOF determination was 2-50 μg mL^-1 with coefficient of determination (R²) > 0.9999. Methods I, II & III were also applied for determination of SOF concentration in single dosage form. Their mean percentage recoveries were 100.35 ± 1.85, 99.97 ± 0.54 and 100.03 ± 0.49, for the three methods respectively. The proposed methods were validated according to international conference of harmonization (ICH) requirements and statistically compared to published reference methods. The ANOVA test confirmed that there is no significant differences between the proposed methods, and can be used for routine analysis of LED and SOF in commercial tablets. These developed methods were applied to estimate the average content and uniformity of dosage unit for LED/SOF combined dosage form and SOF single dosage form according to British pharmacopeia (BP) requirements.