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Chromatography
Tomáš Mlčoch
Pavel Moťka
Chromatography
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Described by Tswett in 1906
He separated some pigments using a
tube filled with CaCO3
By washing the extract through the
column with an organic solvent he was
able to produce several colored bands
band = pás
column = sloupec
Chromatography
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Analytical method
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Separation of components of a mixture
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Stationary phase
– Solid
– Liquid
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Mobile phase (flow over stationary phase)
– Liquid
– Gas
Types of Chromatography
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Column Chromatography
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Thin Layer Chromatography (TLC)
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Paper Chromatography
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Gas Chromatography (GC)
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High Performance Liquid
Chromatography (HPLC)
high performance = vysoce účinná
Column Chromatography
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Vertical glass column
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Mobile phase = liquid
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Stationary phase = solid
– Silica gel (SiO2)
– Alumina (Al2O3)
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Sample = solid or liquid
placed on column
Separatory funnel
Column Chromatography
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Affinity = molecules adsorbed on solid
– Compound in liquid elute first
– More polar molecules elute last
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Collect separated compounds (fractions)
– Evaporate solvent
– ID or further purify
evaporate = vypařit
Thin Layer
Chromatography
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Similar to column chromatography
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Mobile phase = solvent
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Stationary phase = solid (silica gel)
Paper Chromatography
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Similar to TLC
Stationary phase = H2O adsorbed by
cellulose
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Mobile phase = solvent
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Frequently used to polar compounds
– Amino acids, carbohydrates, etc.
Gas Chromatography
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Mobile phase = stream of inert gas
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Stationary phase = high boiling liquid film
– Polarity similar to compounds in mixture
– Supported on a solid
– Packed in a column
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heated glass or metal
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2-3 m in length; 2-5 mm diameter
diameter = průměr
Gas Chromatogram
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Retention time can be used to ID a component if
compared to known sample under same conditions
– Flow rate, column size, temperature, etc.
Resources:
 http://www.labo.cz/sl/vy07.htm
 www.rpi.edu/dept/chem-eng/ BiotechEnviron/CHROMO/chromintro.html
 http://ull.chemistry.uakron.edu/analyti
cal/Chromatography/
 Analytická chemie II, prof. Lumír
Sommer
Thanks