Chromatography.

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Presentation transcript:

Chromatography

The general principle. Use – to separate and identify components of mixtures. Several different types - paper, thin layer, gas-liquid. All use the principle of “partition” - affinity between two phases, to separate mixtures of substances. Stationary phase & mobile phase. Compounds with greatest affinity for mobile phase travel further.

All chromatography needs: support material – stationary phase solvent (or carrier gas) – mobile phase.

Thin Layer Chromatography - t.l.c. Series of spots forms Compare samples in mixture with known substances. Measure Rf values. Coloured compounds & colourless compounds.

Separation and identification.

Gas - Liquid Chromatography G.l.c. Sample introduced by syringe. Column separates components. (Heated in oven) Detector monitors compounds emerging from outlet. Recorder plots signals as a chromatogram.

What happens in practice. Compounds that have high affinity for mobile phase emerge first, (most volatile). Chromatogram charts recorder response against time. Each component - separate peak. Retention time – characteristic of the compound under given conditions.

Factors affecting retention time: length of column packing material type of carrier gas flow rate of carrier gas temperature of column.

Interpreting the trace Calibration – known compounds are added to the column and conditions kept constant. Amount of substance – area under peak / peak height. Relative proportions can be determined.

Uses of G.l.c. Very sensitive - small quantities of substances detected, explosives, drugs etc. Separation of pure substances for collection. Can be connected to mass spectrometer for direct identification of substances.

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