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Applications Of Chromatography Notes

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Applications of Chromatography: Purification and separation Analysis

• E.g. flash chr, preparative

• Qualitative (what present, purity) E.g. TLC. HPLC, affinity chr.

• Quantitative (how much) E.g. gas chr, analytical HPLC.

Thin Layer Chromatography (TLC) Stationary phase: silica gel on inert support. Mobile phase = solvent/solvent mixture, chosen so Rf ≈ 0.3. Separation based on polarity. Polar compounds retained by H bonding to -OHs on gel. Detection of separated compounds by UV, iodine vapour, or ninhydrin (amino acids). Uses: qualitative analysis of small mols, checking purity, monitoring progress of reactions. Gas Chromatography (GC) Stationary phase: high bp, stable liquid, coated onto inside of long, narrow, heated tube. Mobile phase: inert carrier gas, e.g. helium or N2. Only suitable for volatile, thermally stable substances in gas phase.
↑ temperature = ↑ time spent by analytes in gas phase = ↓ retention time. Detection of separated compounds by flame ionisation detectors or mass spectrometer. Flash Chromatography Stationary phase = silica gel. Mobile phase = solvent/solvent mixture, chosen to Rf ≈ 0.2. Pressurized gas used to drive the solvent through the column of stationary phase.

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