INTRODUCTION TO CHROMATOGRAPHY

DR. Harrizul Rivai, m.s.

What is chromatography ?

Based on word “chroma” and “graphein” it mean of color and write

Analysis separation method principle of difference rate of migration sample compound through stationary phase upon mobile phase.

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Chromatography

Stationary Phase

Mobile Phase

Solid or liquid Gas or liquidIn form of In form of

envolving

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Technical Terms in Chromatography :

Stationary phase Liquid or solid which can absorb or hold substance of sample Mobile phase Gas or liquid to move out the substance in sample Elution Separation process of the components in sample

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Technical Terms in Chromatography

Retention time Time needed by analit through the

chromatograph systemChromatogram Picture of chromatography result in

form of peak or spot

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Molecules tend to solve in liquid Molecules tend to stick on the surface of solid Molecules tend to react each other

Trends in chromatography:

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Separated component should be

Soluble in stationary phase

Has ability to interact with the stationary phase by dissolved, adsorbed or chemical reaction.

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Usage of separation result :

Identification (Qualitative analysis )

Content determination (Quantitative analysis )

Purification of substance (Preparative process)

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Advantages of Chromatography :

Small amount sample (semi macro or micro)

Selective, especially for multi component organic substance

Separation process is brief, easy, accurate and precise.

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Chromatography Classification :

1. Based on rate of migration mechanism : adsorption, ion exchange, molecule filtering (exclusion)

2. Based on equipment used : paper, thin layer, column

3. Based on stationary and mobile phase used : gas-liquid, gas-solid, liquid-liquid, liquid-solid.

4. Based on development procedure : frontal analysis, approximation and elusion.

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Types of Chromatography and supporting material

Type Stationary phase Mobile phase Supporting Material

Paper Water Acid or base/ organic substance

Paper

Thin layer Water Acid or base/ organic substance

Thin layer SiO2 orAl2 O3

Column Water Acid or base/ organic substance

SiO2 or Al2 O3 in column

Liquid-Gas(GLC/GC)

Non volatile organic solvent

Gas (N2 or H2) Polymer SiO2 or Al2 O3

Ion exchange Active part of resin (anion/cation)

Reagent /solution Resin

AdsorptionLiquid-Solid / Gas-Solid

Activated Al2 O3 , SiO2

Gas, organic complexing agent

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Molecule Filtration/ exclusion ----------------------

Liquid solvent, gas Solid filter silicate, porous gel

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