|Applicability of retention modelling in hydrophilic-interaction liquid chromatography for algorithmic optimization programs with gradient-scanning techniques
|Comprehensive Chromatography - The State of the Art
| Stef Molenaar
Rianne van Outersterp
|University of Amsterdam
Abstract Information :
Comprehensive two-dimensional liquid chromatography (LCxLC) is an essential technique for the separation of highly complex samples of non-volatile molecules. In principle, the development of an LCxLC method requires establishing two separation dimensions with vastly different ("orthogonal") selectivities. However, with the advent of state-of-the-art instrumentation for LCxLC, the number of options to realize and optimize LCxLC separations is increasing dramatically. The challenge of optimizing tailored LCxLC separations is horrendous, yet it essential to address this challenge if sophisticated LCxLC systems are to be utilized to their full potential in an efficient manner. Method development lengthy and cumbersome and developing methods that make full use of the possibilities of the instrument (e.g. shifting gradients) is barely possible to date.
To facilitate rapid method development of LCxLC methods, the Program for Interpretive Optimization of Two-dimensional Resolution (PIOTR) was developed . It allows rigorous, comprehensive optimization of LCxLC methods by interpreting a very limited number of scouting LCxLC(-MS) experiments. Using gradient-elution theory, large number of possible LCxLC methods are simulated for the interpreted sample, and evaluated using quality descriptors.
However, software tools such as PIOTR require accurate models to describe retention and the exact mechanism and thus model for retention in hydrophilic-interaction chromatography (HILIC) is known to be rather complex. We have evaluated the performance of five different retention models for hydrophilic-interaction chromatography (HILIC) for a wide range of analytes. In this presentation, the results are presented.