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Understanding (and Creating) Polar Retention Using ...
2003 Waters CorporationUnderstanding (and creating ) Polar Retention Using Reversed-Phase HPLC and Hydrophilic Interaction ChromatographyPittcon March 2003 2003 Waters CorporationIntroduction Introduction The Problem Background Hydrophilic Interaction chromatography Reversed-Phase HPLC for Polar Molecules Applications Summary 2003 Waters CorporationIntroduction - The Problem Conventional HPLC columns often do not provide adequate Retention and separation of highly Polar compounds Drug Discovery Gradient - High-throughput screening (HTS) does not work for Polar compounds Elute un-retained in the void volume Co-elute at the beginning of the run if retained at all Method Development Isocratic- Many RP columns cannot be run under the 100% aqueous conditions necessary for Polar compound Retention columns dewet (also termed -hydrophobic collapse) 2003
©2003 Waters Corporation Reversed-Phase Chromatography for Polar Compound Retention •Reversed-Phase Chromatography • Non-polar stationary phase with >80% aqueous mobile phases • Strengths • Familiar, well understood technique • Many stationary phase choices • Good reproducibility, stable equilibration • High efficiency • Weaknesses of modern C 18 phases designed for improved peak
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