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<ul><li>PAPER CHROMATOGRAPHY </li></ul><ul><li>by Mr. Shaise Jacob Faculty, Nirmala College of Pharmacy Muvattupuzha Keral...
PAPER CHROMATOGRAPHY •   ANALYSIS OF UNKNOWN SUSTANCES TWO TYPES 1.PAPER ADSORPTION CHROMATOGRAPHY Paper impregnated with ...
PRACTICAL REQUIREMENTS <ul><li>1)Stationary phase </li></ul><ul><li>2)Application of sample </li></ul><ul><li>3)Mobile pha...
MOBILE PHASE <ul><li>Pure solvents, buffer solutions or mixture of solvents </li></ul><ul><li>Examples- Hydrophilic mobile...
3)ASCENDING – DESCENDING DEVELOPMENT <ul><li>Only length of separation increased, first ascending takes place followed by ...
 
 
<ul><li>DETECTING / VISUALISING AGENTS </li></ul><ul><li>For detecting colorless spots, any one of the following technique...
(b) Specific methods examples.  <ul><li>Ferric chloride </li></ul><ul><li>Ninhydrin in acetone </li></ul><ul><li>Dragendro...
Following detecting tech. can also be categorized as   <ul><li>1) Destructive techniques </li></ul><ul><li>Specific spray ...
APPLICATIONS <ul><li>Separation of mixtures of drugs </li></ul><ul><li>Separation of carbohydrates,vitamins,antibiotics,pr...
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Paper chromatography

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Transcript of "Paper chromatography"

  1. 1. <ul><li>PAPER CHROMATOGRAPHY </li></ul><ul><li>by Mr. Shaise Jacob Faculty, Nirmala College of Pharmacy Muvattupuzha Kerala, India </li></ul>
  2. 2. PAPER CHROMATOGRAPHY • ANALYSIS OF UNKNOWN SUSTANCES TWO TYPES 1.PAPER ADSORPTION CHROMATOGRAPHY Paper impregnated with silica or alumina 2.PAPER PARTITION CHROMATOGRAPHY Moisture / Water present in the pores of cellulose fibers present in filter acts as stationary phase & Mobile phase is used as solvent In general P.C – Paper Partition Chromatography PRINCIPLE OF SEPERATION Partition rather than adsorption Cellulose layers in filter paper contains moisture which acts as Stationary phase & Organic solvents/buffers are used as Mobile Phase
  3. 3. PRACTICAL REQUIREMENTS <ul><li>1)Stationary phase </li></ul><ul><li>2)Application of sample </li></ul><ul><li>3)Mobile phase </li></ul><ul><li>4)Development technique </li></ul><ul><li>5)Detecting or Visualizing agents </li></ul><ul><li>STATIONARY PHASE AND PAPERS USED </li></ul><ul><li>Whatman filter papers </li></ul><ul><li>Paper of any size can be used </li></ul><ul><li>APPLICATION OF SAMPLE </li></ul><ul><li>Capillary tube or Micropipette </li></ul>
  4. 4. MOBILE PHASE <ul><li>Pure solvents, buffer solutions or mixture of solvents </li></ul><ul><li>Examples- Hydrophilic mobile phase & Hydrophobic mobile phases </li></ul><ul><li>DEVELOPMENT TECHNIQUE </li></ul><ul><li>1) ASCENDING DEVELOPMENT (go up) </li></ul><ul><li>FLOWS AGAINST GRAVITY, SPOT KEPT AT THE BOTTOM PORTION OF PAPER & KEPT IN A CHAMBER WITH MOBILE PHASE </li></ul><ul><li>2) DESCENDING TYPE (a downward slope) </li></ul><ul><li>SPECIAL CHAMBER, SOLVENT HOLDER AT THE TOP- SPOT KEPT AT TOP & SOLVENT FLOWS DOWN THE PAPER </li></ul><ul><li>ADVANTAGE IS THAT, DEVELOPMENT IS FASTER </li></ul>
  5. 5. 3)ASCENDING – DESCENDING DEVELOPMENT <ul><li>Only length of separation increased, first ascending takes place followed by descending </li></ul><ul><li>4)CIRCULAR / RADIAL DEVELOPMENT </li></ul><ul><li>Spot is kept at the centre of a circular paper. The solvent flows through a wick at the centre & spreads in all directions uniformly. </li></ul><ul><li>5)TWO DIMENSIONAL DEVELOPMENT </li></ul>
  6. 8. <ul><li>DETECTING / VISUALISING AGENTS </li></ul><ul><li>For detecting colorless spots, any one of the following techniques can be used </li></ul><ul><li>Non specific methods </li></ul><ul><li>E.g. iodine chamber method, </li></ul><ul><li>UV chamber for fluorescent compounds – at 254 or at 365nm. </li></ul>
  7. 9. (b) Specific methods examples. <ul><li>Ferric chloride </li></ul><ul><li>Ninhydrin in acetone </li></ul><ul><li>Dragendroff’s reagents </li></ul><ul><li>3,5 dinitro benzoic acid </li></ul><ul><li>Phenolic comp. & tannins </li></ul><ul><li>Amino acids </li></ul><ul><li>Alkaloids </li></ul><ul><li>Cardiac glycosides </li></ul>
  8. 10. Following detecting tech. can also be categorized as <ul><li>1) Destructive techniques </li></ul><ul><li>Specific spray reagents… </li></ul><ul><li>2) Non-destructive techniques </li></ul><ul><li>For radio active materials , Geiger Muller counter </li></ul><ul><li>QUANTITATIVE ANALYSIS </li></ul><ul><li>Direct techniques- Densitometer </li></ul><ul><li>Indirect techniques- </li></ul><ul><li>QUALITATIVE ANALYSIS </li></ul><ul><li>Rf Value </li></ul>
  9. 11. APPLICATIONS <ul><li>Separation of mixtures of drugs </li></ul><ul><li>Separation of carbohydrates,vitamins,antibiotics,proteins,…. </li></ul><ul><li>Identification of drugs </li></ul><ul><li>Identification of impurities </li></ul><ul><li>Analysis of metabolites of drugs in blood , urine …. </li></ul><ul><li>ADVANTAGES OF P.C </li></ul><ul><li>Simple ,rapid ,inexpensive ,excellent resolving power </li></ul><ul><li>PRECAUTIONS IN P.C </li></ul><ul><li>Establishing the vapor solvent equilibrium </li></ul><ul><li>Stability of solvent mixture is first ensured </li></ul>
  10. 12. Thank you
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