Thin layer chromatography (College)

Category: Education

Presentation Description

No description available.


By: Shyam03 (33 month(s) ago)

HI...could you please send me the ppt about TLC to '' Thank you.

Presentation Transcript

Slide 1: 

Thin Layer chromatography By Abhisek Bhunia

Topic include : 

Topic include

Introduction : 

In 1944, Consden,Gorden & Martin used filter papers for separating the Amino acids. In 1950, Kirchner identified terpenes on filter paper. In 1958,STAHL develop standard equipment for analysing by THIN LAYER CHROMATOGRAPHY . Introduction

Principle : 

The Principle is based on ADSORPTION Chromatography The component with more affinity towards the stationary phase travels slower. The component with lesser affinity towards the stationary phase travels faster. Principle

Reverse phase chromatography : 

In this the stationary phase is Non-polar & mobile phase is polar & it is widely used in pharmaceutical analysis. Polar compound get eluted first Non-polar compounds are retained for long time Comparison of Normal phase & Reverse phase : Reverse phase chromatography

Stationary phase : 

Stationary phase

Mobile phase : 

Nature of the substance to be separated i e weather it is polar or non-polar. Mode of Chromatography. Nature of Stationary phase. Mode Separation i e Analytical or Preparative. Examples: 1) Petroliam ether 2) Cyclohexane 3) Acetone 4) Toluene 5) Ethyl acetate 6) Benzene 7) Alcohols 8) Water 9) Chloroform 10) Pyridine Mobile phase

Glass plates : 

Three types : they are , Full plate : 20cm × 20 cm. Half plate : 20cm × 10 cm. Quarter plate : 20cm × 5 cm. Microscopic slides can also be used for Monitoring the progress of a chemical reaction. Glass plates

Preparation & activation of plates : 

The T L C plates can be prepared by following techniques : Pouring. Dipping. Spraying. Spreading. Activation : It is nothing but removing of water/ moisture & other adsorbed substance from the surface of any adsorbent by heating. Preparation & activation of plates

Application of sample : 

Application of sample

Development tank : 

The development tank should be lined Inside with filter paper moistened with mobile phase to saturate the atmosphere & also prevent the “ EDGE EFFECT ” . Development tank

Development technique : 

Different development techniques are : One dimensional development. Two dimensional development. Horizontal development. Multiple development. Development technique

Detecting agents : 

Detecting agents are two types. they are, Non-Specific method 1) Iodine chamber method. 2) Sulphuric acid spray method. 3) UV chamber for fluorescent compounds. 4) Using fluorescent stationary phase. (B) Specific method 1) Ferric chloride. 2) Ninhydrine in acetone. 3) Dregendroff reagent. 4) 3,5 – Dinitro benzoic acid. 5) 2,4 - Dinitro phenyl hydrazine. Detecting agents

detection : 

The Rf value is calculated for identification "Rf value is the ratio of distance travelled by The solute to the distance travelled by the solvent front” Distance travelled by solute Rf = Distance travelled by solvent front detection

Development of t l c : 

Development of t l c

application : 

Separation of mixture of drug of chemical,biological,plant origin. Separation of Carbohydrates, vitamin, antibiotics, proteins, etc. Identification of drug. Ex :Amoxicillin, Levodopa Detection of foreign substances. To detect the decomposition products of drug. application

Comparison of tlc & paper chromatography : 

Comparison of tlc & paper chromatography

Comparison of tlc & hplc : 

Comparison of tlc & hplc

Reference : 

E Stahl. Thin-layer Chromatography. A Laboratory Handbook: Springer – Verlag Berlin . Heidelberg ,New York 1969:second edition 1990. G Chatwal,S anand.Ihstrumental Methods of Chemical analysis: Himalaya publishing house,Delhi:Edition June 1995. Dr.S. Ravi Sankar.Text book of pharmaceutical analysis:Rx Publication,Tiruneveli:Edition 1997,1999,2001. H.F.Waltor,J.Reyes:Modern Chemical analysis and instrumentation. International Medical Book distrubutor,Mumbai. Thin layer chromatography is available from Thin layer chromatography is available from Thin layer chromatography is available from Thin layer chromatography is available from www.wellesly.eud Reference

Slide 20: