Column chromatography

Column chromatography

1. COLUMN CHROMATOGRAPHY PRESENTED BY RD

2. Introduction History Definition of column chromatography Principle Classification of column chromatography Experimental technique of column chromatography Requirements Application Advantages Disadvantages References Contents

3. DEFINITION :- Chromatography is a physical method of separation in which the components to be separated are distributed between two phases : Stationary phase Mobile phase The stationary phase may be a solid or liquid supported on a solid or gel. May be packed in a column Eg.-Column Chromatography. Spread as a layer on glass/aluminium plate Eg. – TLC, HPTLC. Distributed as a liquid film Eg.- GLC The mobile phase may be a liquid /solvent/mixture of solvent or gases. introduction

4. The first chromatography column was developed by the Russian botanist  Mikhail Tsvet   in 1901. Who washed an organic solution of plant pigments through a vertical glass column packed with an adsorptive material. He discovered that the pigments separated into a series of discrete colored bands on the column, divided by regions entirely free of color. HISTORY

5. Column chromatography is a separation technique in which components of mixture is separated by using a glass column packed with stationary phase and the liquid mobile phase flowing continuously through the column . COLUMN CHROMATOGRAPHY

6. Column chromatography is separated into two categories, depending on how the solvent flows down the column. Gravity column chromatography :- If the solvent is allowed to flow down the column by gravity, it is called gravity column chromatography. Flash chromatography :-If the solvent is forced down the column by positive air pressure, it is called flash column chromatography, currently used in organic chemistry research laboratories Classification of column chromatography

7. A solid stationary phase and a liquid mobile phase is used and the principle of separation is adsorption. The mixture to be separated is dissolved in a suitable solvent and allowed to pass through a tube containing the adsorbent The component which have greater absorbing power is adsorb in the upper part of the column. The next component is adsorb in lower portion of the column which have less adsorbing power than the first component. The process is continued. As a result the materials are partially separated and adsorbed in the various part of the column. The type of interaction between the stationary phase and solute is reversible in nature. PRINCIPLE

8. INSTRUMENTATION TECHNIQUE

9. Column Adsorbents Packing the column Wet packing Dry packing Solvent used Application of the sample Identification and separation of compound Recovery of components Detection of compound REQUIREMENTS FOR COLUMN CHROMATOGARPHY

10. Separation of mixture of compound. Removal of impurities or purification process. Isolation of active constituents. Isolation of metabolites from biological fluids. Estimation of drug in formulation and crude drug extract. APPLICATION

11. Any type of mixture can be separated by column chromatography. Any quantity of mixture can also be. Wider choice of Mobile Phase Advantages

12. Time consuming method. More amount of solvent are required which are expensive. Use of maximum adsorbent. Drying and bubble presence are possible. DISADVANTAGES

13. Kar Ashutosh, “pharmaceutical Analysis’’,CBS Publishers & Distributors pvt.ltd , Volume 2 nd , page no 161-181. Indian Pharmacopoeia , ministry of health sciences government of India 2014 edition , volume 1 st , page no 163-166. REFERENCES