Chromatography LC - LIQUID COLUMN CHROMATOGRAPHY LIQUID...

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LIQUID COLUMN CHROMATOGRAPHY
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LIQUID COLUMN CHROMATOGRAPHY A sample mixture is passed through a column packed with solid particles which may or may not be coated with another liquid. Glass column fused with porous plate/stopper/capillary tubing leading to detector and/fraction collector.
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LIQUID COLUMN CHROMATOGRAPHY Commonly used matrix are: 1. Agarose : polysaccharide made up of D-galactose & 3,6- anhydrogalactose units (gel permeation) 2. Cellulose : polysaccharide of β-1-4 D-glucose units (ion- exchange) 3. Dextran : polysaccharide of α-1-6 D-glucose units (gel permeation) 1.Polyacrylamide : acrylamide polymer (gel permeation) 2.Polystyrene : styrene polymer (ion-exchange) 3.Silica : polymer of orthosilicilic acid (gel permeation)
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LIQUID COLUMN CHROMATOGRAPHY Imp features of matrices are: High mechanical stability High chemical stability Availability (range) ** High capacity (functional groups) Inert surface (avoid non- selective adsorption) ** 1. SEPHADEX G-10 2. SEPHADEX G-100 3. SEPHADEX G-100 SUPERFINE 4. SEPHADEX G-200 5. SEPHADEX G-25 COARSE 6. SEPHADEX G-25 MEDIUM 7. SEPHADEX G-25, FOR GEL FILTRATION 8. SEPHADEX G-50 9. SEPHADEX G-75 10. SEPHADEX LH-20 11. SEPHADEX-DEAE A-25, ION- EXCHANGE RESIN, 40-120 MICRON 12. SEPHADEX-DEAE A-50, ION EXCHANGE RESIN, 40-120 MICRON 13. SEPHADEX-SP, C-25, ION- EXCHANGE RESIN, 40- 120MICRON 14. SEPHADEX-SP, C-50
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LIQUID COLUMN CHROMATOGRAPHY Stationary phases: depending upon requirement Particle size: larger size: faster flow rate, smaller particle: larger surface area, greater resolution Column packing: slurry filled in, no air bubble, flow mobile phase, put nylon gauze on top avoid surface disturbance, always wet.
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