Chemometrically Assisted Development of Ultra-High-Performance Liquid Chromatography Method for the Simultaneous Quantification of Sofosbuvir, Daclatasvir and Ledipasvir in Pharmaceutical Dosage Forms.

Abstract

A new ultra-high-performance liquid chromatography method for the simultaneous quantification of sofosbuvir, daclatasvir and ledipasvir was developed. Two combinations of these direct-acting antivirals are used in hepatitis C virus infection therapy and show high efficacy and safety. Fractional factorial design was used for screening the most influential factors on separation and time analysis. These significant factors were optimized using a central composite design. The optimum resolution was carried out by using a Waters XBridge C18 column (150mm, 4.6mm ID, 5μm) at a temperature of 35°C±2°C and acetonitrile/sodium perchlorate buffer (10mM, pH=3.2) (40: 60v/v) as mobile phase at a flow rate of 1.5mLmin-1. UV detection was set at λ=210nm. A short chromatographic separation time was achieved. The developed method was validated according to the accuracy profile approach and was found specific, precise, faithful and accurate. The detection limits were between 0.07 and 0.13μgmL-1. Hence, this novel method can be employed for the routine quality control analysis and in dissolution profile studies of generics containing these products.