Reliability of simulated robustness testing in fast liquid chromatography, using state-of-the-art column technology, instrumentation and modelling software

• A simulated robustness testing was systematically evaluated.
• Simulated robustness testing and conventional DoE approaches were compared.
• Predicted critical resolution values were experimentally verified.
• Similar selectivity and analysis time could be performed on several columns.
• Modelling softwares could be useful tools in early stage robustness testing.

The goal of this study was to evaluate the accuracy of simulated robustness testing using commercial modelling software (DryLab) and state-of-the-art stationary phases. For this purpose, a mixture of amlodipine and its seven related impurities was analyzed on short narrow bore columns (50 × 2.1 mm, packed with sub-2 μm particles) providing short analysis times. The performance of commercial modelling software for robustness testing was systematically compared to experimental measurements and DoE based predictions. We have demonstrated that the reliability of predictions was good, since the predicted retention times and resolutions were in good agreement with the experimental ones at the edges of the design space. In average, the retention time relative errors were <1.0%, while the predicted critical resolution errors were comprised between 6.9 and 17.2%. Because the simulated robustness testing requires significantly less experimental work than the DoE based predictions, we think that robustness could now be investigated in the early stage of method development.Moreover, the column interchangeability, which is also an important part of robustness testing, was investigated considering five different C8 and C18 columns packed with sub-2 μm particles. Again, thanks to modelling software, we proved that the separation was feasible on all columns within the same analysis time (less than 4 min), by proper adjustments of variables.

Figure optionsDownload as PowerPoint slide