Multivariate Development and Optimization of Stability Indicating Method for Determination of Daclatasvir in Presence of Potential Degradation Products

Abstract

A specific, robust, and rapid UV-HPLC method was developed for the assay of daclatasvir (DCV) in pharmaceutical formulation in the presence of potential degradation products using quality-by-design approach. The impurity profile of DCV was studied via forced degradation procedures with subsequent characterization of the resultant degradation products using LC–MS. Central composite design with response surface methodology was utilized to simultaneously optimize four chromatographic factors: pH, elution temperature, flow rate, and organic modifier %. The first order, interaction, and quadratic effects of those four factors were evaluated for 16 responses of eight peaks “resolution (7), number of theoretical plates (8), run time (1)”. Optimum separation was achieved using Eclipse plus RP C18 column, mobile phase consisting of methanol: 0.025 M phosphate buffer pH 7.0 (58: 42 v/v), flow rate of 1.5 mL min−1at 40 °C, and detection at 303 nm. The optimized method was validated according to ICH guidelines and applied to determine DCV in pharmaceutical formulation. DCV response was linear (r = 0.9999) in the range 1.5–90 µg mL−1; inter-day and intra-day precisions were 0.28% and 0.25%, respectively, and independent t test indicated non-significant difference between inter- and intra-day means; accuracy was 100.49 ± 0.92%.