Validated Stability-Indicating RP-HPLC Method for Daclatasvir in Tablets.

OBJECTIVES

The current study goal was to create a precise, sensitive, and validated reverse phase-high performance liquid chromatography (RP-HPLC) method for assessing the direct-acting antiviral daclatasvir (DCV) as well as to evaluate the stability of DCV in both drug and tablet formulations. The current investigation was to display stability indicating methods under different stress conditions, including hydrolysis (acidic, basic, and neutral), oxidation, and photolysis.

MATERIALS AND METHODS

All experiments were performed on HPLC Agilent 1100 with a stainless steel Hypersil C column having a particle size of 5 μm and a dimension of 4.6 x 250 mm. The mobile phase chosen was acetonitrile: 0.05% o-phosphoric acid (50:50 ) 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, limit of detection, limit of quantification, and robustness in accordance with International Council for Harmonisation (ICH) requirements. The results were satisfactory. It was observed that retention time (t) was 3.760 ± 0.01 min. In acidic conditions, DCV degradans show t at 3.863, 4.121, and 4.783 min and tandem mass spectrometry (MS/MS) spectra scans had 339.1, 561.2 fragment ions. In basic condition, DCV degradans show t at 5.188, 5.469 min and MS/MS spectra scans having 294.1, 339.1, 505.2, 527.2 fragment ions. In oxidation conditions, DCV degradans shows t at 4.038 min and MS/MS spectra scans having 301.1 and 339.1 fragment ions were observed.

CONCLUSION

All the mass fragments exhibited additional degradation observed for different stress conditions. This will help to identify the structure of the degradant and its pathways. No degradation was observed in neutral and photolytic conditions.