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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