The Development of A New Validated HPLC and Spectrophotometric Methods for the Simultaneous Determination of Daclatasvir and Sofosbuvir: Antiviral Drugs

Objective: Simple, rapid and reproducible reversed-phase high performance liquid chromatography (RP-HPLC) and ultraviolet derivative spectrophotometric (UVDS) methods for the simultaneous determination of daclatasvir (DAC) and sofosbuvir (SOF) in pure and in pharmaceutical dosage forms have been developed and validated.

Methods:The chromatographic separation of DAC and SOF was achieved using Agilent Zorbax SB C18 (4.6 x 250 mm, 5 μm) column at temperature of 40 °C. The mobile phase used was 9 mM dipotassium hydrogen orthophosphate buffer (pH 4±0.1): acetonitrile (60:40, v/v). The flow rate was maintained at 1 mL/min with UV detection at 265 nm.

Results: The calculated resolution was 4.56 (> 2), which ensures complete separation. The tailing factor was 1.13 and 1.40 (≤ 2) for DAC and SOF, respectively. Intermediate precision value was ≤ 2% indicate acceptable ruggedness.

Conclusion: The two methods were validated according to the International Conference on Harmonization (ICH) guidelines in terms of linearity, precision, accuracy, selectivity, specificity, detection limit, quantification limit, robustness and ruggedness.

Author(s): Amira S. Eldin, Shereen M. Azab, Abdalla Shalaby, Magda El-Maamly