Types of Liquid Chromatography.ppt

Types of Liquid Chromatography I. Ion Exchange Chromatography A. Factors influencing retention B. Suppressed ion exchange II. Partitioning Chromatography A. Normal phase/ reverse phase III. Size Exclusion Chromatography IV. Supercritical Fluid Chromatography/ SFE V. Capillary Electrophoresis Factors Influencing retention in Ion Exchange Ionic strength: not real important in selectivity pH: anion exchange pH % retention % cation exchange pH % retention Temperature: T % efficiency % Flow rate: Slightly slower than other HPLC methods to maximize resolution improve mass transfer kinetics Suppressed Ion Chromatography Partitioning Chromatography Analyte interacts with mobile and stationary phase, differential interaction leads to selectivity Interactions that are important Proton accepting ability * most important Dipole interaction Proton Donor * most important e- pair donating ability Van der Waals dispersion forces Types of Partitioning Chromatography Types of Partitioning Chromatography Solvophobic Theory Water has a lot of intermolecular interactions in the liquid phase Solute dissolved in water disrupts those intermolecular interactions Solute is forced out of aqueous phase not because of favorable interactions between analyte and stationary phase but because of unfavorable interactions between solute and water when solute is dissolved in aqueous phase hence: SOLVOPHOBIC THEORY Polar functional groups such as –OH would increase the favorability of interaction and thus decrease retention (in mobile phase longer) Polar things elute non-polar things elute Size Exclusion Chromatography Size Exclusion Chromatography Size Exclusion Chromatography Summary Supercritical Fluids Supercritical Fluid Chromatography Instrumentation Properties of Mobile Phases Used in Chromatography Supercritical fluid extraction (SFE) Supercritical fluid extraction Insert Hawthorne paper * * Buffer salt: Influences pH selectivity Organic Modifier: Solvent strength increase