column chromatography ppt.pptx

column chromatography ppt.pptx

• 1.
  Column Chromatography Presented to:Dr Pervez Anwar Presented By: Hassan yousaf Roll no : 20204022-014 Presentation title: Column Chromatography Department: MS Biological sciences University of Sialkot
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  ⦿Useful method forthe separation and purification of both solids and liquids . ⦿Solid – liquid technique ⦿stationary phase – solid ⦿Mobile phase – liquid
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  Principle ⦿Adsorption ⦿ Mixture ofcompounds dissolved in mobile phase is introduced into the column . ⦿Separation occurs because of the different rates at which the components are carried over the stationary phase by the mobile phase
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  ⦿Adsorption column chromatography, the adsorbent packed in glass column . ⦿Solvent – mobile phase , moves slowly through the packed column . ⦿Process of dissolving out the components from the adsorbent using a suitable solvent is called elution . ⦿This Solvent used as mobile phase – eluent .
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  ⦿Compound attracted morestrongly by the mobile phase will move rapidly through the column . ⦿ Compound more strongly attracted to the stationary phase will move slowly through the column
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  Stationary phase OFCOLUMN CHROMATOGRAPHY ⦿ADSORBENTS : silica ,alumina, calcium carbonate , calcium phosphate , magnesia and starch ⦿Alumina suitable for chromatography of less polar compounds . ⦿Silica gel suitable for compounds containing polar functional groups .
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  DIFFERENT MOBILE PHASE USED ⦿Petroleumether ⦿ Carbon tetrachloride ⦿Cyclohexane ⦿ Ether ⦿ Ester ⦿ Acetone ⦿ Benzene ⦿Water
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  PREPARTION OF COLUMN ⦿Consists ofglass tube with bottom portion of the column – packed with cotton / glass wool ⦿Above which adsorbent is packed . ⦿After packing , paper disk is kept on the top , so that the adsorbent layer is not disturbed during the introduction of mobile phase .
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  PACKING TECHNIQUES OF ADSORBENTS ⦿DRYPACKING ⦿WET PACKING THE PROCESS ⦿ An adsorbent column is prepared by packing a long glass tube with suitable adsorbent ⦿solution of the mixture of the components (say a,b,c) in suitable solvents is introduced at the top of the column .
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  ⦿ It percolatesthrough the adsorbent column , different components adsorbed to different extend . ⦿ Most readily adsorbed constituent(say a) is held at the top . ⦿ Other (say b and c) of decreasing adsorbabilities are held up at different zones or bands down the column in the same order [say b and then c] ⦿ This partial separation is improvised by adding some amount of original solvent – development of chromatogram.
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  ⦿Individual components areextracted from the adsorbent by a suitable solvent(eluent). ⦿Most weakly adsorbed component (c) emerges first which is at the bottom of the column . ⦿Most strongly adsorbed one (a ) comes out last which was at the top of the column . ⦿Pure components are then recovered by removing the solvent by suitable method .
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  APPLICATIONS ⦿Used for separationof two or more similar organic components of a mixture ⦿Identification , separation and purification of natural products. ⦿Used for identification of various industrial products ⦿Separation of protein mixture.
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  Advantages of CC ⦿Anytype and any quantity of mixture can be separated . ⦿Wider choice of mobile phase DISADVANTAGES OF CC ⦿Time consuming ⦿ More amount of mobile phase is required .
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