Strategy for developing and optimizing liquid chromatography methods in pharmaceutical development using computer-assisted screening and Plackett–Burman experimental design

摘要: We describe a three-step method development/optimization strategy for HPLC assay/impurity methods pharmaceuticals, which include multiple-column/mobile phase screening using system equipped with column-switching device, further optimization of separation by multiple organic modifiers in the mobile phase, and multiple-factor Plackett-Burman experimental designs. In first two steps, commercially available chromatography software, DryLab, was used to perform computer simulations. This allows developer evaluate each condition (one column/mobile combination) retention data from scouting gradient runs. approach significantly reduces number runs development. After satisfactory obtained, we step The purpose 16-injection set experiments nine factors regard precision, accuracy, sensitivity specificity. results provided logical justifications selecting parameters such as column temperature, detection wavelength, injection volume, sample solvent, etc. analysis, instead traditional mathematical manipulations, graphical examine present creating so-called main effect plots. Because replicates design points were not run, did allow testing statistical significance. However, it visual presentations way that is easy understand end user alike.