Please cite this article in press as: Baker MM, et al. Validated spectrophotometric and chromatographic methods for analysis of the recently approved hepatitis C antiviral combination ledipasvir and sofosbuvir. Ann Pharm Fr (2017), http://dx.doi.org/10.1016/j.pharma.2017.07.005 ARTICLE IN PRESS +Model PHARMA-520; No. of Pages 16 Annales Pharmaceutiques Françaises (2017) xxx, xxx—xxx Disponible en ligne sur ScienceDirect www.sciencedirect.com ORIGINAL ARTICLE Validated spectrophotometric and chromatographic methods for analysis of the recently approved hepatitis C antiviral combination ledipasvir and sofosbuvir Méthodes spectrophotométriques et chromatographiques validées pour l’analyse de la combinaison antivirale de l’hépatite C récemment approuvée, le ledipasvir et le sofosbuvir M.M. Baker a , D.S. El-Kafrawy b , M.S. Mahrous b , T.S. Belal c,* a Methodology Department, Pharco Pharmaceuticals Company, Alexandria, Egypt b Pharmaceutical Chemistry Department, Faculty of Pharmacy, University of Alexandria, Elmessalah 21521, Alexandria, Egypt c Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, University of Alexandria, Elmessalah 21521, Alexandria, Egypt Received 13 May 2017; accepted 24 July 2017 KEYWORDS Ledipasvir; Sofosbuvir; Derivative spectrophotometry; Ratio spectra; HPTLC; HPLC-DAD Summary This work describes five simple and reliable spectrophotometric and chromato- graphic methods for analysis of hepatitis C antiviral binary mixture of ledipasvir (LPV) and sofosbuvir (SBV). Method I is based on the use of A max and derivative spectrophotometry with the zero-crossing technique where LPV was determined using its A max and 1 D amplitudes at 324 and 338 nm respectively, while SBV was determined by measuring the 1 D amplitudes at 276 nm. Method II involves the application of the ratio spectra derivative spectrophotometry. For LPV, 12 g/mL SBV was used as divisor and the 1 DD amplitudes at 239.8 nm were plotted against LPV concentrations; while by using 10 g/mL LPV, the amplitudes at 279.2 nm were found propor- tional to SBV concentrations. Method III depends on ratio-difference measurement where the ∗ Corresponding author. E-mail address: tbelaleg@yahoo.com (T.S. Belal). http://dx.doi.org/10.1016/j.pharma.2017.07.005 0003-4509/© 2017 Acad´ emie Nationale de Pharmacie. Published by Elsevier Masson SAS. All rights reserved.