Testing experimental designs in liquid chromatography (II): Influence of the design geometry on the prediction performance of retention models

Abstract

In liquid chromatography, the reliability of predictions carried out with retention models depends critically on the quality of the training experimental design. The search of the best design is more complex when gradient runs are used instead of isocratic experiments. In Part I of this work (JCA 1624 (2020) 461180), a general methodology based on the error propagation theory was developed and validated for assessing the quality of training designs involving gradients. The treatment relates the mathematical properties of a retention model with the geometry of the training designs and their subsequent predictions. In that work, only five usual designs were considered. Part II investigates in detail the effects on predictions when the features of the training design (number and distribution of the experiments, initial and final modifier content, gradient slope(s), and location of gradient nodes and pulses) are varied systematically. Several groups of related designs containing one or more isocratic steps, linear or multi-linear gradients, or mixed isocratic/gradient runs, among others (in total 38 designs) were evaluated. Box and whiskers and triple plots of expected relative uncertainties were used to evidence the differences in prediction performance. The purpose was to give recommendations to construct designs with good prediction performance. The best designs sample (considering all runs) concentrations as diverse as possible, at any gradient time.