Chromatography
• Chromatography is a separation technique in which
sample components distributes themselves between
two immiscible phases (mobile phase and stationary
phase) to varying degrees. Consequently, they migrate
at different rates through the S.P. under the influence
of the M.P.
What is
Chromatography?
A+B
signal
tr(B)
tr(A)
A
to
B
Time
A
B
Glossary of chromatographic terms
• Stationary phase– Phase that stays intact.
– Usually a viscous liquid coated on the inside of a
capillary tube or on the surface of solid particles
– May contain minerals, organic or inorganic polymers or
liquid-coated solids
• Mobile phase
– Phase that moves.
– May be gases, liquids or supercritical fluids.
• Columns
– Usually contains the stationary phase
– Either packed or open tubular
– Packed columns are filled with particles of the stationary
phase
– The stationary phase is coated on the inside of the
column in open tubular columns
• Eluent
- Fluid entering the column
• Effluent
– Fluid leaving the column
• Elution
– Process of passing a liquid or gas through a chromatography
column
Elution order
• Compounds that have a low affinity for the column packing
and/or a high solubility in the mobile phase are washed
through the column quickly.
• Compounds that stick tightly to the column packing and/or
have a low solubility in the mobile phase solvent move
more slowly.
• Developing an HPLC separation requires finding a
combination of column packing and mobile phase that
provides just the right balance of affinity and solubility to
make the compounds of interest move at different speeds.
Classification of chromatography
• according to the types of M.P and S.P
Chromatography
Gas Chromatography
GLC
GSC
Liquid Chromatography
LLC
LSC
• M.P :mobile phase, S.M: stationary phase,
IEC
SCF Chrom.
SEC