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CHEMISTRY 1000. Chromatography. Chromatography. Chromatography is a technique for separating species based on physical or chemical properties. Usually made up of the species to be separated, a stationary phase, and a mobile phase.
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CHEMISTRY 1000 Chromatography
Chromatography • Chromatography is a technique for separating species based on physical or chemical properties. • Usually made up of the species to be separated, a stationary phase, and a mobile phase. • Separation of different compounds depends on relative affinity of the compounds for the stationary phase and the mobile phase.
Ion-Exchange Chromatography • Separates ions based on magnitude of charge (must all have the same sign of charge. • Stationary phase usually consists of beads, whose surfaces are covered with charged groups. Examples: • The mobile phase is a salt solution of progressively increasing concentration. • Ions with larger charges are more strongly attracted to the stationary phase and can displace ions with smaller charges. • To remove ions with largest charge, flush system with conc. solution of another ionic compound.
Ion-Exchange Chromatography • ion-exchange chromatography – the experiment.
Liquid Chromatography • Separates ions based on polarity. • Two techniques: • Normal phase: stationary phase is a polar solid while the mobile phase is a less polar liquid. example: silica as stationary phase and ether/hydrocarbon mixture as mobile phase • Reverse phase: stationary phase is nonpolar while the mobile phase is more polar.
Liquid Chromatography • More polar solutes are more strongly attracted to stationary phase (stronger IMF). • Less polar solutes are less attracted to the stationary phase (weaker IMF). • Therefore less polar species will travel faster through the column than more polar species. • For given experimental conditions, the time for a species to elute (move through the column) is highly reproducible. • High performance liquid chromatography (HPLC) is a widely used type of liquid chromatography in which the mobile phase is pressurized to push it through the stationary phase more quickly.
Gas Chromatography (GC and/or GC/MS) • Stationary phase is a viscous liquid coated on the inside of a thin tube called a column. • Mobile phase is an inert gas. • Separates ions based on boiling point (IMF): • substances with higher boiling point spend more time in the stationary phase (stronger IMFs result in longer retention in the stationary phase). • boiling point increases with stronger IMFs. • Molecules in the gas can become dissolved in the stationary phase, slowing their progress through the column.
Gas Chromatography (GC and/or GC/MS) • Chromatography is used to separate mixtures. • When used with Mass Spectrometry (MS) this can be a powerful technique to separate (purify) and identify compounds. • Applications: • Environmental analyses • Identifying illicit substances • Explosives detection • Medical applications