Systematic, computer-assisted development of high performance liquid chromatography for multi-component analysis

Abstract

The aim of this study was to develop a multi-component determination analytical method. The analytical Quality by Design (QbD) concept was used in the beginning of the establishment of a high performance liquid chromatography (HPLC) method for compound traditional Chinese medicine (TCM) preparations using a diode-array detector (DAD) and an evaporative light scattering detector (ELSD) in series. Herein, the QbD workflow is discussed and demonstrated with a systematic HPLC method development, including risk assessment, the design of experiments (DOEs), and assessment of the data to provide a method operable design space (DS). Modeling software Drylab was employed to set up experiments for the development of a simple and robust separation method and to visually achieve the required criteria as an initial DS of the analytical method based on simulation. To improve the method development and optimization step, the statistical software JMP@(SAS Institute) was applied to simultaneously optimize the chromatographic conditions such as the gradient time, the concentration of the aqueous phase, the column temperature, the flow rate and the ELSD parameters. Finally, a successful HPLC method was developed and validated to verify the robustness of the QbD system. The use of QbD workflows streamlines the development of methods as compared to traditional approaches. With the addition of systematic DOEs, the optimization resulted in critical resolution R_s, crit ≧ 1.5 for all the six compounds researched. As a result, a robust and reliable method operable design region was established. This method had fewer issues and failures throughout the lifecycle due to the knowledge gained via the QbD process.