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.

signal

Time

BA

tr(A)

to

tr(B)

A+B

A B

What is Chromatography?

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

Liquid Chromatography

Chromatography

SCF Chrom. Gas Chromatography

GLC GSC LSCLLC IEC SEC

• according to the types of M.P and S.P

• M.P :mobile phase, S.M: stationary phase,