Application of QbD based approach in development and validation of RP-HPLC method for simultaneous estimation of Pregabalin and Naringin in Dual-Drug Loaded Liposomes.

The current work delineates the development of a novel rugged & sensitive stability indicating risk-based HPLC method based on an analytical Quality-by-Design (QbD) approach for the concurrent estimation of Naringin (NRN) and Pregabalin (PRG) in dual-drug loaded nanopharmaceuticals. Preliminary screening trials were conducted, along with systemic risk analysis, in order to identify the critical method attributes, namely injection volume, pH, and acetonitrile content, that influence critical quality attributes. The Box-Behnken design (BBD) was used to optimise the tailing factor as a response to pregabalin and naringin in a short run time. The chromatographic conditions were improved by running 17 experimental runs generated by design expert software. After analysing the optimised zone within the confines of the design space, the following chromatographic conditions were chosen: mobile phase water: acetonitrile adjusted to pH 6.9 with phosphate buffer (80:20, %v/v), at flow rate of 1.0 ml/min using an analytical column C18 at an isobestic wavelength of 212 nm. Furthermore, the optimised method was validated in accordance with ICH guidelines and was found to be within the prescribed limits. The developed RP-HPLC method has a high degree of practical utility in invivo and invitro studies for synchronous detection of Pregabalin and Naringin in pharmaceutical nano-dosage forms such as protein-based nanoparticles, nanocrystals, polymeric nanoparticles, metallic nanoparticles, etc.