SC 3 - HPLC Method Development and Optimization

PD Dr. Frank Steiner, Dionex Corporation, Dornierstraße 4, 82110 Germering, Germany
Dr. Stefan Lamotte, Bischoff Analysentechnik und -geräte GmbH, Leonberg, Germany

Course contents:
Method development is still considered a crucial bottleneck in analytical laboratories and impedes their productivity to a significant extent. To cope with this challenge, it is recommended to pursue clearly defined rules and procedures. This enables even inexperienced users to develop adequate methods in a minimum amount of time. Starting from properties of the sample, we elucidate simple guidelines to select the best suited column and elution conditions. Different retention mechanisms like e.g.; normal phase, reversed phase, ion pair and hydrophilic interaction liquid chromatography are introduced and illustrated to explain the related background. We present a systematic strategy to find the ideal stationary / mobile phase combination. This mainly involves a combinatorial approach regarding all the above mentioned variable parameters, experimentally executed by generic gradient test runs. Either manual or fully automated scouting of such combinations will lead to a set of promising methods. Method optimization and simulation software is briefly discussed as helpful tools for straight forward fine tuning of method parameters, e.g. multi segment gradient profiles. This section of the workshop also addresses method robustness optimization.
A further improvement of a so found initial method might be possible through kinetic optimization mainly based on the van Deemter theory. Recent technologies like e.g.; UHPLC on sub-2-µm materials, fused core particles and phase optimized liquid chromatography are discussed. We present method speed-up calculators and other software tools to assist such optimization steps, especially for phase optimized liquid chromatography. This section also covers instrumental requirements and rigid precautions that the operators have to consider in order to achieve robust UHPLC analyses. The seminar will be in an open and interactive style and involve every participant’s individual input to the maximum possible extent.

Instructors:
Frank Steiner studied chemistry at Saarland University in Saarbruecken, Germany. He received his Ph.D. in 1995 in the group of Prof. Dr. Dr. h.c. Heinz Engelhardt for a thesis entitled "Processing and Monitoring of Wastewater by Ion Chromatography". After a postdoc from 1996-1997 at the Centre d’Études Nucléaires de Saclay, France, he was an assistant professor at Saarland University and got qualified as an associate professor in 2003. In 2005 he moved to Dionex Corporation, Germering, Germany, where he is a Product Manager for HPLC instrumentation and a specialist for small molecule analysis solution development.

Stefan Lamotte studied chemistry at Saarland University in Saarbruecken, Germany. He received his Ph.D. 1998 in the group of Prof. Dr. Dr. h.c. Heinz Engelhardt. His thesis was about synthesis, characterization and application of 1.5 µm nonporous particles for fast HPLC. Since 1998 he is manager of columns and stationary phases at Bischoff Analysentechnik und -Geraete GmbH in Leonberg, Germany. Dr. Lamotte is a specialist in silica chemistry, stationary phases for HPLC, and phase-optimized liquid chromatography.