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Chromatography - Coggle Diagram
Chromatography
Paper
Chromatography
Rf value
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measured
- from base line
- to centre of spot
method
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2.small spots of mixture
- using capillary tube
- repeat = conc spot
3.place in tank
- bottom of paper touch solvent
- allow run
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STATIONARY phase
- water bonded to the cellulose
- in the paper
Two-Way
Chromatography
used to separate AMINO ACIDS
why?
- aa have similar structure
- ∴ different solvents separate more completely
all amino acids = colourless
- ∴ sprayed with ninhydrin - stains aa
OR
- use UV light light + trace round w. pencil
method
1. draw origins using a pencil
2. spot mixture onto paper
- using capillary tube
- repeat = conc spot
- allow to dry
3. place paper in tank
- allow to run in first solvent
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- chromatogram run 1 way in 1 solvent
- DRIED
- placed ANOTHER solvent + run
Thin-Layer
Chromatography
STATIONARY phase
- silica/alumina
- on glass/plastic backing plate
spots detected using...
UV light
- spots fluoresce
- marked with pencil
Iodine Vapour
- TLC placed in container w/ iodine vapour
- adheres to spots & marks them
Ninhydrin
- for amino acids
- stains purple
method
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3.place in sealed beaker
- stops ethanol evaporating
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6.place TLC
- under UV light
- in iodine
- spray ninhydrin
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silica coating = POLAR environment
- if mobile phase = NON-POLAR
- spend most time in mobile phase = high Rf
- if mobile phase = POLAR
- spend most time in stationary phase = low Rf
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