Validated Stability-indicating RP-HPLC Method for Daclatasvir in Tablets

INTRODUCTION: The current study goal was to create a precise, sensitive and validated RP-HPLC method for assessing the direct-acting antiviral daclatasvir (DCV) as well as to evaluate the stability of the DCV in both drug and tablet formulations. The current investigation was to perform stability indicating method in different stress conditions, including hydrolysis such as (acidic, basic and neutral), oxidation and photolysis.
METHODS: All the experiment was performed on HPLC Agilent 1100 having stainless steel Hypersil C18 column with a particle size of 5µm and a dimension of 4.6 X 250 mm. The mobile phase chosen was acetonitrile: 0.05% o-Phosphoric acid (50: 50 v/v) in isocratic mode with 0.7 mL/min flow rate and wavelength 315 nm was selected for detection.
RESULTS: This method was validated for linearity and range, accuracy, precision, LOD, LOQ and robustness in accordance with ICH requirements. The results were satisfactory. It was observed that retention time (tR) was 3.760±0.01 min. In acidic condition, DCV degradant shows tR at 3.863, 4.121, 4.783 min and MS/MS spectra scans having m/z 339.1, 561.2 fragment ion. In basic condition, DCV degradant shows tR at 5.188, 5.469 min. and MS/MS spectra scans having m/z 294.1, 339.1, 505.2, 527.2 fragment ions observed. In oxidation conditions, DCV degradant shows tR at 4.038 min and MS/MS spectra scans having m/z 301.1, 339.1 fragment ions were observed.
DISCUSSION AND CONCLUSION: All the mass fragments shows the additional degradation observed for different stress conditions. This will help to identify the structure of degradant and its pathways. No degradation was observed in neutral and photolytic conditions.