Column & short column chromatography (priyansha singh notes chapter 3)

Unit 3 separation techniques Column & Short Column Chromatography
1. Basic Principles
2. Column packaging
3. Column development
4. Detection

1. COLUMN & SHORT COLUMN CHROMATOGRAPHY UNIT 3, SEPARATION TECHNIQUES (NP- 510) PRIYANSHA SINGH (PC2021-14/226) M.S. PHARM- PHARMACOLOGY & TOXICOLOGY NIPER- GUWAHATI

2. CONTENTS BASIC PRINCIPLES COLUMN PACKING COLUMN DEVELOPMENT DETECTION

3. HISTORY & BASIC CONCEPT 

4. TYPES OF COLUMN CHROMATOGRAPHY

5. STATIONERY PHASE- aka adsorbent/ solid. MOBILE PHASE- aka solvent & eluent/ liquid. SAMPLE- aka adsorbate which gets adsorbed. ELUTION- process of removing the components from column. ELUATE- separated component. TERMINOLOGY

6. Adsorption Chromatography Stationery phase- solid Mobile phase- liq./ gas PRINCIPLE OF ADSORPTION CHROMATOGRAPHY

7. Rate of movement of component in adsorption chromatography

8. STATIONERY PHASE

9. TYPES OF ADSORBENTS

10. SELECTION OF ADSORBENT

11. PREPARATION OF STATIONERY PHASE

12. DEVELOPERS

13. MOBILE PHASE To introduce mixture into the column as solvent. To develop the zones for separation as developing agent. To remove pure component out of the column as the  Eluant. Different mobile phases uses for e.g.- in increasing order of polarity/ elution strength Cyclohexane< Carbondisulfide< ether< benzene< toluene< esters< alcohols< chloroform< acetone< water< pyridine< organic acids.

14. COLUMN CHARACTERISTICS

15. PACKAGING OF COLUMN Click to add text

16. Importance of packing

17. TYPES OF PACKAGING     DRY PACKAGING                                                       WET PACKAGING

18. INTRODUCTION OF SAMPLE

19. ELUTION GRADIENT/ STEPWISE ELUTION The solvents of increasing polarity/ elution strength are used during the process of separation. Initially low polar solvent is used followed by the increasing polarity e.g.- initially benzene then chloroform, ethyl acetate, methanol etc. ISOCRATIC GRADIENT ELUTION In this elution technique, the same solvent/ solvent system of same polarity is used throughout the process of separation. e.g.- chloroform only, petroleum: ether= 1:1

20. Elution contd..... ONE CARTIRIDGE CAN SEPARATE ALL 3 DYES

21. DETECTION OF COMPONENTS IN SAMPLE Click to add text

22. RECOVERY OF COMPONENTS IN SAMPLE 

23. APPLICATIONS

24. FACTORS AFFECTING COLUMN EFFICIENCY

25. ADVANTAGES v/s DISADVANTAGES of COLUMN CHROMATOGRAPHY ADVANTAGES Any type of mixture can be separated by this method. Any quantity of mixture can also be separated. Wider choice of mobile phase In preparative type sample can be separated & reused. Automation is possible                 DISADVANTAGES Time consuming More amount of solvents are required which are expensive Automation makes the technique more complicated.

26. Partition Column Chromatography GRADIENT ELUTION MEANS TO MAINTAIN A VARIABLE CONCENTRATION IN THE MOBILE PHASE.

27. SHORT COLUMN CHROMATOGRAPHY A chromatography method where the stationary bed is within a tube (of less than the standard length of 25 cm). The particles of the solid stationary phase or support coated with a liquid stationary phase may fill the whole inside volume of the tube (packed column) or be concentrated on or along the inside tube wall leaving an open, unrestricted path for the mobile phase in the middle part of the tube (open-tubular column).