Chapter 37 Analytical Techniques (Chromatography) Flashcards
Thin-layer Chromatography & Gas/liquid Chromatography
What is chromatography used for?
- to separate & analyse small amounts of mixtures
- uses the principle of a mobile phase (liquid or gas) moving past a stationary phase
Define mobile phase.
- allows molecules to move through the stationary phase
- can be liquid or gas
- more soluble products move further with the mobile phase
Define stationary phase.
- a substance that has affinity to molecules in the mixture being analysed
- the greater the affinity of a molecule to the stationary phase, the shorter the distance it moves with the mobile phase
Two mechanisms of chromatography.
- separation by partition
- separation by adsorption
Chromatographic separation by partition. State:
- stationary phase
- mobile phase
- how components separate
- stationary phase - liquid film on an inert surface
- mobile phase - liquid or gas
- components separate according to solubility in each phase
Chromatographic separation by adsorption. State:
- stationary phase
- mobile phase
- how components separate
- stationary phase - solid
- mobile phase - liquid or gas
- components separate according to strength of bond with the stationary phase
The 4 types of chromatography.
- Paper chromatography
- High Pressure Liquid Chromatography
- Thin-layer chromatography (in syllabus)
- Gas Liquid Chromatography (in syllabus)
Rf value formula.
= distance moved by solute / distance moved by solvent
- Calculated Rf values are compared with reference values obtained under identical conditions (e.g. same temperature & same solvent used).
Thin-layer chromatography. State:
- stationary phase
- mobile phase
- mechanism
- method of separation
- stationary phase - thin layer of silica gel or alumina coated on to a piece of glass, metal or plastic to make it rigid
- mobile phase - organic solvent that moves up the TLC plate by capillary action
- mechanism - adsorption
- method of separation - Components in a mixture are separated based on their different abilities to bind or adsorb to the stationary phase, & on their different abilities to desorb or dissolve in the mobile solvent phase.
Advantages of TLC.
- faster
- inexpensive
- achieves better separations (due to greater choice in stationary & mobile phases)
- works with very small samples
- separates a wide range of mixtures
How does silica gel cause the separation of solutes in TLC?
- the surface of silica gel is very polar
- because of -OH groups, can form Hydrogen bonds, London Dispersion Forces & Dipole-dipole attractions with certain compounds
What happens to polar molecules with a polar stationary phase in TLC?
- Polar molecules have a greater attraction for a polar solid & are adsorbed more strongly onto its surface.
- They travel more slowly up the thin layer of alumina or silica & separation occurs.
- Solutes are located on chromatogram & identified by comparing Rf values.
Uses of TLC.
- Identification of biological sugars
- Identification of components that might damage paper like -COOH(s)
- Separation of insecticides, steroids
Gas liquid chromatography. State:
- stationary phase
- mobile phase
- mechanism
- stationary phase - high boiling point non-polar liquid on an inert solid support
- mobile phase - inert gas (helium, nitrogen)
- mechanism - partition
Method of separation of GLC.
- Small amount of sample injected into the machine.
- Injector is contained in an oven.
- Sample boils & is carried along a thin column by an inert carrier gas.
- Column contains a liquid stationary phase, adsorbed onto an inert solid.
- Time taken to travel through the tube will depend on how much time is spent moving with the gas rather than being attached to the liquid.