column chromatography.pptx

chromatography, types, column chromatography, working principle, applications

1. Column chromatography Submitted by ; A.Suruthi I M.sc Biotechnology Under the guidance Dr . R. Kalaivani Assistant Professor of Biotechnology Bon Secours College for Women, Thanjavur

2. Column chromatography Introduction : A chromatography column is a device Used in chromatography For the separation of chemical compounds. A chromatography column The stationary phase, allowing the Mobile phase to pass though it. chromatography columns of different types are Used in both Gas and liquid chromatography.

3. Column chromatography Column chromatography Was developed by The American chemist D.T day In 1990, M.S tswett, the polish botanist, in 1906 used adsorption Columns In his investigations of Plant Pigments.

4. Column chromatography olumn chromatography is one of most useful Methods for The seperation and purification of both solids and liquids. This is a solid-liquid technique in which the Stationary phase Is a solid and mobile phase is a liquid.

5. Column chromatography . Principle Adsorption Mixture of Components Dissolved In the M.P is Introduced In to the column. Components moves depends upon their relatives affinities. Adsorbtion Column Chromatography, The Adsorbent, Packed in a glass column, and a solvent, The mobile phase, That movies Slowly Through The Packed Column A Solvent Used as a Mobile phase Is called an elunt.

6. Column chromatography

7. Column chromatography Stationary phase Success Of chromatography depends Upon Proper Selection of S.P, it depends following : 1.Removal Of Impurities. 2. No.of components to be seperated. 3.Length of the column used. 4.Affinities difference between Components. 5.Quentity Of adsorption Used.

8. Column chromatography Mobile phase The act as Solvent, develaper and eluent. The function of a mobile phase are : 1.As developing agent 2.To introduce the mixture into the column-as Solvent. 3. To developing agent. 4.To remove pure components Out of the column – as eluent.

9. Column chromatography Packing technique in Column Chromatography there are two types of preparing The column They are: 1. Dry packing / dry filling 2. Wet packing / wet filling . The column Should be free form impurity before using Column it shoud be washed properly and dry it. Before Filling Column with Stationary Phase, cotton / glass Wool Is kept. it should be Uniformly filled.

10. Column chromatography Dry packing Technique : Adsorbent is backed in the column in dry form Fill the solvent, Till Equilibrium Is reached. Ofter filling Tapping Can be tone to Remove void Spaces.

11. Dry packing Technique

12. Column chromatography Wet packing technique : Ideal and common technique: the meterial is Slurried With solvent and generally added to the Column in portions. S.P Settles Uniformly And no crack In the Column of Adsorbent . Solid Settles down while the solvent remain Upward. This solvent is removed then again cotton plug is placed.

13. Column chromatography Application: Separation of mixture of Components. Purification Process. Isolation of active Constituent. Estimation of drugs in formulation. Determination of Primary and secondary glycosides in digitalis Leaf. Sepration of diasteromers.

14. Column chromatography Advantages : Any type of Mixture can be separated. Any quantity mixture can be seperated. Wider choice of mobile phase. Automation is possible.

15. Column chromatography Disadvantages : time Consuming. More amount of Mobile phase as requested. Automation Makes the technique More complicated And expensive.