CHROMATOGRAPHY (DEMONSTRATION)

CHROMATOGRAPHY
Chromatography basically involves the separation of mixtures due to differences in the distribution coefficient (equilibrium distribution) of sample components between 2 different phases which are immiscible. One of these phases is a mobile phase and the other is a stationary phase.

porous medium through which the mobile phase

migrates is called the support.

Distribution Coefficient (Equilibrium Distribution )

Definition: Concentration of component in stationary phase

Concentration of component in mobile phase

Different affinity of these 2 components to stationary phase causes the separation.

DUE DIFFERNCE IN DISTRIBUTION COEFFICIENT OF VARIOUS COMPONENT OF MIXTURE, THERE MOBILITY IN TWO PHASES DIFFER HENCE, SEPARATION MEASURE OF THIS MOBILITY IS CALLED RF VALUE I.E. RELATIVE FRACTION VALUE

Distance traveled by substance

Rf=

Distance traveled by solvent

STATIONARY PHASE: IN SOME CASES SOLID SUPPORT IT SELF FORM THE STATIONARY PHASE , WHILE IN OTHERS , IT ABSORBS LIQUID PHASE WHICH IN TURN ACTS AS STATIONARY PHASE. IT IS SOLID OR LIQUID.

MOBILE PHASE: PHASE WHICH MOVE OVER OR THROUGH STATIONARY PHASE AND CARRIES SAMPLE ALONG WITH IT, THUS RESULTING IN THE SEPARATION OF ITS COMPONENT. IT IS EITHER LIQUID OR GAS.

Kinds of Chromatography
1. Liquid-Liquid Column Chromatography

Paper Chr., Gel filteration, Ion exchange, Affinity, Column Chromatography.

2. Liquid-Gas Chromatography GC 3. Solid- Liquid Chromatography

TLC, HPTLC

 Important Terminology: Adsorption Partition

Elution
Eluent Chromatogram Chamber Saturation Detection Affinity

 Principles of Chromatography: Adsorption Phenomena

Adsorption chromatography is probably one of the oldest types of chromatography around. It utilizes a mobile liquid or gaseous phase that is adsorbed onto the surface of a stationary solid phase. The equilibriation between the mobile and stationary phase accounts for the separation of different solutes.
Examples- TLC, HPTLC, CC

Partition Phenomena
This form of chromatography is based on a thin film formed on the surface of a

solid support by a liquid stationary phase. Solute equilibriates between the mobile phase and the stationary liquid. Examples- PLC, HPLC, GC, CC