Thin layer chromatography

ATUL CHAUDHARY (STUDENTS)
DEPARTMENT OF PHARMACEUTICS
ISF COLLEGE OF PHARMACY, GHALKALAN, MOGA , PUNJAB
THIS SLIDE IS THE BEST SLIDE FOR PREPARING THE TOPIC THIN LAYER CHROMATOGRAPHY OF M.PAT SUBJECT
SLIDE ARE SPECIALLY DESIGN FOR MASTER STUDENTS AS WELL AS GRADUATION STUDENTS

1. THIN LAYER CHROMATOGRAPHY Seminar presentation ( I.S.F. COLLEGE OF PHARMACY MOGA) Presented by- Atul Chaudhary M.PhaRm . First Year PHARMACEUTICS

2. What is chromatography Chromatography is a laboratory technique for the separation of a mixture. The mixture is dissolved in a fluid called the mobile phase ,  which carries it through a structure holding another material called the stationary phase. The separation is based on differential partitioning between the mobile and stationary phases Subtle differences in a compound's partition coefficient result in differential retention on the stationary phase and thus affect the separation 12/1/2019 2 T.L.C. ASSIGNMENT

3. WHAT IS T.L.C. Thin-layer chromatography (TLC) is a widely employed laboratory technique used to separate different biochemicals on the basis of their relative attractions to the stationary and mobile phases. TLC is very versatile; multiple samples can be separated simultaneously on the same layer, making it very useful for screening applications such as testing drug levels and water purity. 12/1/2019 3 T.L.C. ASSIGNMENT

4. Principle of T.L.C. 11/30/2019 4 T.L.C. ASSIGNMENT

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6. Plate Prepration and activation The slurry, which is a mixture of stationary phase and water is prepared by using the ratio mentioned earlier. After preparing the slurry, the TLC plates can be prepared by using any one of the following techniques. Pouring Dipping Spraying Spreading 11/30/2019 6 T.L.C. ASSIGNMENT

7. Conte... Normal thickness of 0.25cm is used for analytical purpose and 2mm thickness for preparative purpose Then the spreader is rolled only once on the plate. The plate are allowed for setting . This is done to avoid cracks The plates are activated by keeping in an oven at 100 degree centegrade to 120 degree centigrade for 1 hour 11/30/2019 7 T.L.C. ASSIGNMENT

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9. Spotting 2 – 5 of 1% solution of sample or standard is spotted using a capillary tube or micropipette. The spots should be kept at least 2cm above the base of plates and the spotting area should not be immersed in mobile phase in a developing chamber. The sample is applied on the narrow bands The width of the band must be as narrow as possible 11/30/2019 9 T.L.C. ASSIGNMENT

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11. How to Run T.L.C Step 1: Prepare the developing container Step 2: Prepare the TLC plate Step 3: Spot the TLC plate Step 4: Develop the plate Step 5: Visualize the spot 11/30/2019 11 T.L.C. ASSIGNMENT

12. How to Calculate Rf value 11/30/2019 12 T.L.C. ASSIGNMENT

13. Some visualising Agents UV Light:  For aromatics + conjugated systems. Iodine:  Visualizes ~half the time. Anisaldehyde :  For many aldehydes , ketones , and alcohols. Vanillin:  For many aldehydes , ketones , and alcohols. Permanganate:  For alkenes, alkynes, or oxidizable groups ( aldehydes , alcohols). Phosphomolybdic Acid (PMA):  For alcohols, phenols, alkenes, and many carbonyl compounds. Iron(III) Chloride:  For phenols. Bromocresol Green:  For acidic compounds 11/30/2019 T.L.C. ASSIGNMENT 13

14. Applications of TLC TLC is more useful for the analysis of polar compounds like Amino acids, Sugars, Natural products etc. The Different types of applications are listed below Seperation of mixture of drugs of chemical or biological origin, plants extracts etc, Seperation of carbohydrates (sugar) Vitamins, proteins, alcohals , glycosides, aminoacids etc 11/30/2019 14 T.L.C. ASSIGNMENT

15. Some Other Applications Identifications of drugs Identifications of related compounds in drugs To detect the presence of foreign substance in drugs To detect decomposition products in drugs 11/30/2019 15 T.L.C. ASSIGNMENT

16. Conte.. 11/30/2019 16 T.L.C. ASSIGNMENT

17. REFERENCES Bernard Fried and Josef Sherma Prem kumar Jaiswal Egon Stahi Chatwal Google Wikipidia Slideshare 11/30/2019 T.L.C. ASSIGNMENT 17

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