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CHROMATOGRAPHY
 
Methods in Experimental PhysiologyAugust 31 & September 1, 1999Dr. Stojiljkovic
CHROMATOGRAPHY
PARTITION CHROMATOGRAPHY
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Gel-filtration chromatography
 
ADSORPTION CHROMATOGRAPHY
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Ion-exchange chromatography
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Affinity chromatography
 
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY(HPLC)
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High performance normal phase chromatography
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High performance reverse phase chromatography
 
 
 GELS USED IN CHROMATOGRAPHY TECHNIQUES:
Term gel refers to fairly soft elastic material containing water. A gel consistsof three-dimensional structure. A structural material, often consisting of cross-linked polymers, gives a gel a mechanical stability. The space withinthe gel not occupied by structural material is filled with liquid.Important properties of gels for chromatography1. The matrix of the gel must be inert.2. The gels must be stable chemically.3. Low content of ionic groups is required to avoid ion exchange effects.4. The mechanical rigidity of the gel grains should be a high as possible.
 Dextran gels
Dextran is a polysaccharide, built up from glucose residues.1. Sephadex2. Sepahcryl is a dextran gel manufactured by cross-linking. Thecross-reaction gives a gel the very high rigidity.
 Polyacrylamide gels
They are prepared by co-polymerization of acrylamide and N,N-methylen-bis-acrylamide.
 Agarose gels
Agarose gels are made of mixture of linear polysaccharides composedmainly from D-galactose and anhydro-L-galactose residues.
 
 
TECHNIQUES OF PARTITION CHROMATOGRAPHY:GEL FILTRATION
 
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Gel filtration is a technique of partition chromatography in whichthe partitioning is based on the molecular size of the substances tobe separated.
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The partition coefficient of the solute between the gel phase andthe liquid phase is governed exclusively by steric effect.
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Large molecules cannot penetrate into the beads and thus willappear from the column first. The small molecules can pass intothe gel beads and lag behind the large molecules.
 Applications of gel filtration chromatography
1. Analytical applications
a. Analytical group separationb. Analytical fractionationc. Determination of molecular masses
2. Preparative applications
a. Preparative group separationsb. Preparative fractionation
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mariecurie85

I couldn't download the pdf. how can ı download your papper? thanks for your attention.

reply05 / 01 / 2009
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