A Quality by Design (QbD) Framework for Reversed-Phase Liquid Chromatography Method Development

Abstract

A framework for reversed-phase HPLC method development was developed by combining computer simulation, UPLC scouting experiments and surface response design. An interactive computer simulation program was developed for optimizing the gradient elution conditions. This simulation program integrated the kinetics and flow resistance parameters of 18 commonly used columns. Accurate prediction of retention times by this simulation program was demonstrated in this work. This program was used to propose the optimum gradient operational conditions under given constraints, which greatly reduced the time required for method development. The Box-Behnken design was used to evaluate the optimum operational conditions obtained from the simulation and experiments and define the method operable design space. Further insights into the separation mechanisms were obtained from the statistical analysis results of DOE.