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CHROMATOGRAPHY
Ferosekhan . S
FNB-41
Chromatography
Chromatography is a technique for separating mixtures
into their components in order to analyze, identify, purify,
and/or quantify the mixture or components.
• Analyze
Separate • Identify
• Purify
Mixture Components
• Quantify
Mechanisms Of Separation
Gas chromatography
Liquid chromatography
Column Chromatography
Planar Chromatography
solvent front
component B
B. Dissolve solid
sample in MeOH
B. Developing solution
is drawn up the plate
by capillary action
Mobile phase:
acetonitrile : water (85:15)
Detection:
sulfuric acid : methanol (1:3)
heat for 10 min at 110 C to see
brown spots
Separation of Total Lipid into
different Classes
Cholesteryl esters
TAG
Cholesterol
1,3-DAG
1,2-DAG
Monoacyl glycerols
Phospholipids
Tristearin
2-oleodistearin
1-stereodiolein
Triolein
Trolinolein
With HUFA
Uses
Separation
Identification
Chromatography paper
Stationary phase
Solvent
Mobile phase
Paper Chromatography
Cont…
Paper chromatography is a variant of partition
chromatography procedure in which the
cellulose support is in the form of a sheet or
paper
2 mm
1 cm
Cont…
Spot and label each of the four indicators and one
of the unknowns.
The spots should be about 2 cm apart.
When the spots have dried, re-spot each one.
2 cm
Cont…
When the spots have dried, form the
paper into a cylinder with the spots
facing out. Staple the edges together
being careful to keep them straight
and not allowing them to touch.
Separation of carbohydrates: