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Chromatography Techniques Tutorial

Key Concepts

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Comparison of Chromatography Techniques

Type Features Uses
Paper

  • inexpensive
  • slow
  • often used for water soluble compounds
  • a qualitative technique
  • stationary phase: usually water (present in the paper);
    mobile phase:pure solvent or solvent mixture
  • Retardation factor, Rf, measured

  • identification of colours in inks or foods
  • identification of amino acids
  • separation of pigments in plant leaves

Thin Layer (TLC)

  • inexpensive
  • faster than paper and more sensitive
  • used for less polar compounds
  • qualitative technique although components may be scraped off the plate & their masses measured
  • stationary phase: plate coated with silica gel or Al2O3;
    mobile phase:pure solvent or solvent mixture
  • Retardation factor, Rf, measured

  • identification of biological sugars
  • identifying components of pain-relieving preparations
  • identifying components (eg, carboxylic acids) that might damage paper

Gas
(GC, GLC)
  area under the peak measures the amount of the component present
  • expensive
  • fast
  • used for gases or easily vaporised compounds
  • relative molecular mass (Mr) < 300
  • both qualitative and quantitative technique
  • stationary phase: a liquid adsorbed onto an inert solid;
    mobile phase (carrier gas): an inert gas such as helium, nitrogen
  • Retention time, Rt, measured
  • analysis of air-borne pollutants
  • analysis of athletes' urine samples for performance enhancing drugs
  • analysis of oil spills
  • analysis of essential oils in perfume preparation

High-performance Liquid
(HPLC)
  area under the peak measures the amount of the component present
  • expensive but less expensive than gas chromatography (GC)
  • fast but slower than gas chromatography (GC)
  • used for organic compounds that decompose if vaporised or compounds with relative molecular mass (Mr) > 300
  • both a qualitative and a quantitative technique
  • stationary phase: small waxy solids;
    mobile phase (eluent): pure solvent or solvent mixture
  • Retention time, Rt, measured
  • analysis of pharmaceuticals
  • cyanide analysis
  • analysis of organic pollutants in water
  • analysis of toxic compounds in shell fish
  • protein analysis

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