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.


Amira S. Eldin, Shereen M. Azab, Abdalla Shalaby, Magda El-Maamly