Chromatography technique_Presentation.pptx

Chromatography is a separation technique used to separate and identify components of a mixture.

1. Chromatography

2. Introduction Chromatography is a separation technique used to separate and identify components of a mixture. It is based on differential distribution between stationary and mobile phases. Discovered by Mikhail Tswett in 1906.

3. Principle Separation occurs due to differences in affinity towards stationary and mobile phases. Higher affinity for stationary phase → slower movement. Higher affinity for mobile phase → faster movement.

4. Types of Chromatography 1. Based on Phase Type: - Adsorption Chromatography - Partition Chromatography 2. Based on Mobile Phase: - Gas Chromatography (GC) - Liquid Chromatography (LC) 3. Based on Technique: - Column, TLC, Paper, Ion Exchange, Gel Filtration, HPLC

5. Column Chromatography Stationary phase: Solid (silica gel, alumina) Mobile phase: Liquid solvent Used for isolation and purification of compounds.

6. Thin Layer Chromatography (TLC) Stationary phase: Thin layer of silica gel. Mobile phase: Solvent mixture. Steps: 1. Apply sample spot 2. Develop plate 3. Visualize under UV/iodine Rf = distance by solute / distance by solvent front

7. Paper Chromatography Stationary phase: Water in cellulose fibers. Mobile phase: Organic solvent. Types: Ascending, Descending, Radial. Used for amino acids, sugars, drugs.

8. Ion Exchange Chromatography Separation based on charge. Stationary phase: Ion exchange resin. Types: Cation & Anion exchange. Used for purification of proteins and amino acids.

9. Gel Filtration Chromatography Also called Size Exclusion Chromatography. Stationary phase: Porous gel beads. Larger molecules elute first. Used for protein and polysaccharide separation.

10. High Performance Liquid Chromatography (HPLC) Modern column chromatography using high pressure. Stationary phase: Silica/polymer. Mobile phase: Solvent under pressure. Applications: Drug analysis, purity testing, quality control.

11. Applications • Separation and purification of drugs • Identification of compounds • Quantitative and qualitative analysis • Detection of impurities • Used in clinical and biological research

12. Advantages • High sensitivity and accuracy • Small sample size required • Applicable to wide range of compounds • Rapid and reliable results

13. Limitations • Expensive instruments (HPLC, GC) • Requires skilled personnel • Limited for non-volatile or thermally unstable compounds (in GC)

14. Summary Chromatography is essential in pharmaceutical and biochemical studies. Choice of method depends on sample and purpose. It plays a vital role in drug development and quality assurance.

15. References 1. Chatwal & Anand – Instrumental Methods of Chemical Analysis 2. Pavia – Introduction to Organic Laboratory Techniques 3. Beckett & Stenlake – Practical Pharmaceutical Chemistry