Chromatography

A) Precautions
1- Free of adulteration. Avoid any chemical changes
2- Diseased part removed. occurring before extraction.
3- Inactivated enzymes.
B) Extraction
Depends on: 1) texture, water content
2)substance
Methods:
Cold Hot/total Soxhelt Supercritical fluid
Proced. plt+solv+stiring Plt+ethanol/aq.alc+ Plnt+cont.reflux Gas+Press+powder
at r.t heat+reflux+filter Flask,condenser +remove press
Advan. -suitable with -suit. With -big yields -clean extract
labile. lipophilic/polar -low amt. of solv. - polarity by
-moderate -drug discovery methanol or
extractiondue to CH2Cl2
equilibrium bet. -monoterpines
soln.and plant mat.
Disadv. -may deactivate -degrade labile -has chemical resd N.B:
bio.active drugs. - may deactivate -oxidation All solvents are
-monoterpines bio.active drugs. -resinification acc. To Eluotropic
insoluble in polar -monoterpines - may deactivate Series.
compounds insoluble in polar bio.active drugs.
compounds -temp. stressed

Types:
1- Hydrophilic: extraction by polar solvents
2- Hydrophobic: extraction by non-polar solvents.
C) Concentration
Method Rotatory evaporator Blown down Freeze dried
Char. >5ml 1:5ml Remove of water as ice
Under vacuum Under N2 Using lyophilizer

D) Storage
- At -20 c.
- Possibility of bioactive natural product degradation by
enzymes.
 E) Identification
1- Partial (test, solubility, Rf, uv spectro, biochemically)
2- Complete(melting, boiling, optical rot, Rf to stand., all spectro,
authentic mat., x-ray crystallography)
F) Classification
Acc.to Mobile phase Stationary Separation mech. Equipment, operational
procedures
Types -liquid - Gas-solid -Adsorption A)column
-supercritical fluid - liquid-liquid -partition -column
-gas - gas-liquid -ion exchange resin -HPLC
-affinity in bio fluids -GC
-gel filteration B)TLC
C)paper
Partitioning
Characters:
1- Soft method no heat
2- 2 immiscible org. solvents
3- Initial extract purification
4- Depends on solubility
Theory:
- Distribution of solute between 2 immiscible solvents is an
equilibrium condition.
- K=[Sorg]/[Saq]
- K is equilibrium constant, partition coefficient, distribution
coefficient.
Method:
- From non-polar : medium : polar solvents.