THIN LAYER CHROMATOGRAPHY.pptx
- 1. THIN LAYER CHROMATOGRAPHY SUBMITTED BY GOPIKA MS-22MFS10019 SREELAKSHMI S-22MFS10010
- 2. CONTENTS THIN LAYER CHROMATOGRAPHY PRINCIPLE PREPARATION OF TLC PLATE STEPS IN TLC CALCULATION OF Rf VALUE APPLICATION OF TLC CONCLUSION
- 3. CHROMATOGRAPHY Chromatography is a process of separation in which the components to be separated are distributed between two immiscible phases a stationary phase and mobile phase.
- 4. THIN LAYER CHROMATOGRAPHY Thin layer chromatography (TLC) is an important technique for identification and separation of mixtures of organic compounds . TLC can be used to help determine the number components in a mixture, the identity of compounds ,and purity of a compound. TLC is performed using thin sheets of glass ,aluminium or plastic coated with a layer of stationary phase usually silica gel is used.
- 5. PRINCIPLE It is based on the principle of adsorption chromatography or partition chromatography or combination of both ,depending on the adsorbent, its treatment and nature of solvents employed. The components with more affinity towards stationary phase travels slower. Components with less affinity towards stationary phase travels faster.
- 6. PREPERATION & ACTIVATION OF TLC PLATE Pouring (simplest method ) Dipping (used for small plates ) Spraying( difficult to get uniform layers) Spreading( best technique)
- 7. STEPS IN TLC TLC consist of three steps Spotting Development Visualization
- 8. SPOTTING In TLC the samples must be carefully applied to the plate to minimize spreading. Samples sizes from 0.1 mg to 50mg are the best for TLC. Smaller amount are difficult to visualize, while the larger spots result in over loaded and variable results. Sample should be dissolved in relatively volatile solvents.
- 9. DEVELOPMENT The spotted plate is placed in a sealed development tank filled with vapour of mobile phase . Its lower side immersed in the solvent to a level below the applied sample spots. The solvent rises due to capillary flow in a process called development. Development times can range from 3-60 minutes .
- 10. VISUALIZATION Visible-the bands or spots can be seen immediately. Fluorescence –observed under UV light. Reaction with a chromogenic reagent .
- 11. CALCULATION OF Rf VALUE The basic parameter used to describe migration in TLC is the Rf value where, Rf=Distanced travelled by solute Distance travelled by solvent Rf value vary from 1 to 0
- 12. APPLICATION OF TLC To check the purity of given samples. Identification of compounds like acids ,alcohols, proteins etc . Used in cosmetic industry. It is used for testing of Drug. Ink analysis.
- 13. CONCLUSION TLC is the method used for analyzing mixtures by separating the compounds in the mixture .On a TLC plate there is a competition between the stationary phase , silica gel and the mobile phase ,ethyl acetate's can be used to help determine the number of components in a mixture, to identify the compounds ,and the purity of a compound.
- 14. THANK YOU