Column chromatography by kk sahu sir

Introduction
Definition
History
Types of chromatography
Principle of column chromatography
Types of column chromatography
Process of column chromatography
Requirement
Procedure
Precautions
Applications
Advantage of Column chromatography
Disadvantage of Column chromatography
Conclusion
References

1. COLUMN CHROMATOGRAPHY       By KAUSHAL KUMAR SAHU Assistant Professor (Ad Hoc) Department of Biotechnology Govt. Digvijay Autonomous P. G. College Raj-Nandgaon ( C. G. )

2. Synopsis Introduction Definition History Types of chromatography Principle of column chromatography Types of column chromatography Process of column chromatography Requirement Procedure Precautions Applications Advantage of Column chromatography Disadvantage of Column chromatography Conclusion References    

3. Introduction It is a broad range of physical methods used to separate and or to analyze complex mixtures A mixture of molecules is separated based on each molecules degree of affinity for the mobile and stationary phase.  Column chromatography is the actual or basically a type of adsorption chromatography techniques

4. Definition Column chromatography is one of the most useful methods for the separation and purification of both solids and liquids. This is a solid - liquid technique. History American chemist D.T. Day in 1900. Invented in 1906 by the Russian botanist Mikhail Tsvet.

5. Types of chromatography Gas chromatography Liquid chromatography High performance Liquid chromatography Ion exchange chromatography Column chromatography Thin layer chromatography Paper chromatography.

6. Principle of Column Chromatography When a mixture of mobile phase and sample to be separated are introduced from top of the column. The individual components of mixture move with different rates. Those with lower affinity and adsorption to stationary phase move faster and eluted out first while those with greater adsorption affinity. slower and get eluted out last.

7. Two phase present in column chromatography Stationary phase Mobile phase The stationary phase = It is a solid. The most common stationary phase is silica gel. An important ratio between the stationary phase weight and the dry weight of the analyte mixture.

8. The mobile phase It is either a pure solvent or a mixture of different solvents. It is liquid form. There is an optimum flow rate for each particular separation

9. Types of column chromatography Adsorption chromatography Partition chromatography Ion exchange chromatography Gel chromatography Adsorption Chromatography It is one of the oldest types of chromatography . Utilizes a mobile liquid or gaseous phase.

10. The equilibriation between the mobile and stationary phase accounts for the separation of different solutes

11. Partition Chromatography This form of chromatography is based on a thin film formed. Solute equilibriates between the mobile phase and the stationary liquid.

12. Ion Exchange Chromatography In this type of chromatography, the use of a resin. resin is the stationary solid phase covalently attach anions or cations onto it. To the opposite charge in the mobile liquid phase are attracted to the resin by electrostatic forces

13. Molecular Exclusion Chromatography Also known as gel permeation or gel filtration An attractive interaction between the stationary phase and solute

14. The liquid or gaseous phase passes through a porous gel which separates the molecules according to its size. Process of column chromatography Requirements Cylindrical column Adsorbent Solvent Cotton wool

15. Procedure The stationary phase material is packed sufficiently in the column with a cotton pad at the bottom. The extract material to be separated is placed on the top of packed stationary phase with a second cotton pad in between. The mobile phase is poured into the column over the sample. A collecting beaker to collect the elute.

16. Precautions: Keep the column in a clean and dust free place. Do not disturb the column till the separation is complete. Avoid gaps within the stationary phase packing

17. Applications To separate or purify natural compound mixtures like alkaloids, glycosides. Column chromatography is best suited to separate active principle from plant materials. Advantage s of Column Chromatography Any type and quantity of mixture can be separated Wider choice of Mobile Phase Automation is possible

18. Disadvantages of Column Chromatography Time consuming More amount of Mobile Phase are required Automation makes the techniques more complicated & expensive.

19. Conclusion Chromatography is a very special separation process for a multitude of reasons. It can separate complex mixtures with great precision. The main advantage of column chromatography is the relatively low cost. Disposability of the stationary phase used in the process.

20. Reference Principle of biochemistry5th edition- David L.Nelson Michael M.Cox. www.studymode.com(at10 pm 9 se p) http://en.wikipedia.org

21. Thank you