This is a draft cheat sheet. It is a work in progress and is not finished yet.
Chromatography
Group of techniques used to separate complex mixtures on basis of different physical interactions between a stationary and mobile phase |
Basic components |
- mobile phase (gas or liquid) |
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- stationary phase (solid or liquid) |
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- column holding stationary phase |
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- separated components (eluate) |
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- complex mixture (sample) |
Forms |
- Planar |
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- Paper |
Column |
- Gas (GC)- can be mobile or stationary (polymer silica) |
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- Liquid (LC) – mobile phase – high pressure and connected to mass spec |
Modes of Separation |
- Adsorption – liquid-Solid (hydrophobic and Hydrophilic bonds) |
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- Partition – Liquid-Liquid |
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- Steric exclusion – Variation of liquid-Solid |
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- Ion-exchange chromatography – Magnitude and charge (separation of amino acids) |
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- Affinity |
CHROMATOGRAPHIC PROCEDURES
Thin-layer chromatography (TLC) |
- A variant of column chromatography |
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- Thin layer of sorbent is uniformly coated on glass/plastic plate. |
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- Samples are applied as spots near edge of plate. |
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- One edge of plate is placed in solvent. |
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- Solvent migrates up thin layer by capillary action. |
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- After solvent reaches predetermined height, plate is removed and dried. |
HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
- Uses pressure for fast separations, controlled temperature, in-line detectors, and gradient elution techniques |
- Components: pumps, columns, sample injectors, detectors, recorders |
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