IB Chemistry on Paper Chromatography, Thin Layer and Column Chromatography
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IB Chemistry on Paper Chromatography, Thin Layer and Column Chromatography

IB Chemistry on Paper Chromatography, Thin Layer and Column Chromatography

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IB Chemistry on Paper Chromatography, Thin Layer and Column Chromatography IB Chemistry on Paper Chromatography, Thin Layer and Column Chromatography Presentation Transcript

  • Tutorial on Chromatographic Techniques,Paper Chromatography , Thin Layer and ColumnChromatography. Prepared by Lawrence Kok http://lawrencekok.blogspot.com
  • Chromatography Techniques Chromatography Techniques • Separation technique of mixture into their pure components • Identify if sample is mixture or pure both quantitatively and qualitatively • Interaction of substances between 2 phases - Stationary phase and Mobile phase • Separation is based on Partition or Adsorption Separation analysis ChromatographyPartition Chromatography Adsorption Chromatography
  • Chromatography Techniques Chromatography Techniques • Separation technique of mixture into their pure components • Identify if sample is mixture or pure both quantitatively and qualitatively • Interaction of substances between 2 phases - Stationary phase and Mobile phase • Separation is based on Partition or Adsorption Separation analysis Chromatography Partition Chromatography Adsorption Chromatography Partition Chromatography Adsorption Chromatography• Components distribute bet TWO immisible liquid phase • Component adsorb on solid stationary phase• Depends on relative solubility bet TWO phase • Depends on polarity of stationary phase, mobile phase and solutes• Solutes bonds to stationary phase or mobile phase • Stationary phase is polar – polar solutes adsorb strongly • Stationary phase is non polar – non polar solutes adsorb strongly • Mobile phase is polar – solutes stay in mobile phase • Mobile phase is non polar – non polar solutes stay in mobile phase
  • Chromatography Techniques Chromatography Techniques • Separation technique of mixture into their pure components • Identify if sample is mixture or pure both quantitatively and qualitatively • Interaction of substances between 2 phases - Stationary phase and Mobile phase • Separation is based on Partition or Adsorption Separation analysis Chromatography Partition Chromatography Adsorption Chromatography Partition Chromatography Adsorption Chromatography• Components distribute bet TWO immisible liquid phase • Component adsorb on solid stationary phase• Depends on relative solubility bet TWO phase • Depends on polarity of stationary phase, mobile phase and solutes• Solutes bonds to stationary phase or mobile phase • Stationary phase is polar – polar solutes adsorb strongly • Stationary phase is non polar – non polar solutes adsorb strongly • Mobile phase is polar – solutes stay in mobile phase • Mobile phase is non polar – non polar solutes stay in mobile phase Paper Chromatography Thin Layer Chromatography Column Chromatography Y adsorb strongly X adsorb strongly Application Application Application • Detection of amino acids in a mixture • Detection of amino acids in a mixture • Preparative/collection of sample • Different dyes in food colouring • Different dyes in food colouring of pigments • Separation of plant pigments • Separation of plant pigments
  • Chromatography Techniques ChromatographyPartition Chromatography Adsorption Chromatography• components distribute between 2 immisible liquid phase • components adsorp on solid stationary phase• relative solubility in 2 phase• bonds strongly to mobile phase – move faster Stationary Liquid phase Stationary phase O- Stationary phase has a layer of liquid O- • solid • AI2O3 O- • SiO2 O- O-
  • Chromatography Techniques ChromatographyPartition Chromatography Adsorption Chromatography• components distribute between 2 immisible liquid phase • components adsorp on solid stationary phase• relative solubility in 2 phase• bonds strongly to mobile phase – move faster Stationary Liquid phase Mobile Y Mobile Y Y Stationary phase liquid phase X O- Stationary phase liquid phase X has a layer of liquid Y • solid containing containing Y O- • AI2O3 X and Y X X and Y X Y O- • SiO2 Y X X O- O-
  • Chromatography Techniques ChromatographyPartition Chromatography Adsorption Chromatography• components distribute between 2 immisible liquid phase • components adsorp on solid stationary phase• relative solubility in 2 phase• bonds strongly to mobile phase – move faster Stationary Liquid phase Mobile Y Mobile Y Y Stationary phase liquid phase X O- Stationary phase liquid phase X has a layer of liquid Y • solid containing containing Y O- • AI2O3 X and Y X X and Y X Y O- • SiO2 Y X X O- O- Mobile Mobile O- Y Y Y adsorb strongly XX Y Stationary phase Iiquid phase XX Y has a layer of liquid liquid phase O- Y containing X Y containing X XX Y X X O- Y X X
  • Chromatography Techniques ChromatographyPartition Chromatography Adsorption Chromatography• components distribute between 2 immisible liquid phase • components adsorp on solid stationary phase• relative solubility in 2 phase• bonds strongly to mobile phase – move faster Stationary Liquid phase Mobile Y Mobile Y Y Stationary phase liquid phase X O- Stationary phase liquid phase X has a layer of liquid Y • solid containing containing Y O- • AI2O3 X and Y X X and Y X Y O- • SiO2 Y X X O- O- Mobile Mobile Y Y O- Y adsorb strongly XX Y Stationary phase Iiquid phase XX Y has a layer of liquid liquid phase Y O- containing X Y containing X XX Y X X Y O- X X X X X X Separation of X and Y X Y Y Y X X YYY Y
  • Chromatography Techniques ChromatographyPartition Chromatography Adsorption Chromatography• components distribute between 2 immisible liquid phase • components adsorp on solid stationary phase• relative solubility in 2 phase• bonds strongly to mobile phase – move faster Stationary Liquid phase Mobile Y Mobile Y Y Stationary phase liquid phase X O- Stationary phase liquid phase X has a layer of liquid Y • solid containing containing Y O- • AI2O3 X and Y X X and Y X Y O- • SiO2 Y X X O- O- Mobile Mobile Y Y O- Y adsorb strongly XX Y Stationary phase Iiquid phase XX Y has a layer of liquid liquid phase Y O- containing X Y containing X XX Y X X Y O- X X X X X X Separation of X and Y X Y Y Y X X YYY YPartition –distribution of solute X and Y between 2 liquid phase Adsorption– solute X and Y adsorb temporary on the solid• X more soluble in mobile phase (move with mobile liquid phase) • Y adsorb strongly on solid phase, eluted slower• Y less soluble in mobile phase (stay on stationary liquid phase) • X in liquid mobile phase, eluted faster
  • Chromatography Techniques Paper Chromatography Thin Layer Chromatography Partition chromatography Adsorption chromatography • Distribution solute bet both liquid phase • Solute adsorb on stationary solid phase • Depends on relative solubility O- Stationary phase solid Stationary phase - Cellulose paper X O- • AI2O3 or SiO2 • absorb water on its surface Y Y Y O- X Y O- Y Y X O- X Mobile Liquid phase with solute X and YMobile Liquid phase with solute X and Y
  • Chromatography Techniques Paper Chromatography Thin Layer Chromatography Partition chromatography Adsorption chromatography • Distribution solute bet both liquid phase • Solute adsorb on stationary solid phase • Depends on relative solubility O- Stationary phase solid Stationary phase - Cellulose paper X O- • AI2O3 or SiO2 • absorb water on its surface Y Y Y O- X Y O- Y Y X O- X Mobile Liquid phase with solute X and YMobile Liquid phase with solute X and Y Aqueous liquid phase on surface of stationary phase (paper) Stationary phase - solid silica SiO2 or alumina AI2O3 Mobile phase – liquid solvent move by capillary action Mobile liquid phase - solvent Separation depends on how strongly solute adsorb on solid Solvent move by capillary action Adsorption of solute on stationary solid phase
  • Chromatography Techniques Paper Chromatography Thin Layer Chromatography Partition chromatography Adsorption chromatography • Distribution solute bet both liquid phase • Solute adsorb on stationary solid phase • Depends on relative solubility O- Stationary phase solid Stationary phase - Cellulose paper X O- • AI2O3 or SiO2 • absorb water on its surface Y Y Y O- X Y O- Y Y X O- X Mobile Liquid phase with solute X and YMobile Liquid phase with solute X and Y Aqueous liquid phase on surface of stationary phase (paper) Stationary phase - solid silica SiO2 or alumina AI2O3 Mobile phase – liquid solvent move by capillary action Mobile liquid phase - solvent Separation depends on how strongly solute adsorb on solid Solvent move by capillary action Adsorption of solute on stationary solid phase
  • Chromatography Techniques Column Chromatography Thin Layer Chromatography Adsorption chromatography Adsorption chromatography• Solute adsorb on stationary solid phase • Solute adsorb on stationary solid phase Mobile Liquid phase (solvent) with solute X and Y X Stationary phase solid O- Stationary phase solid X X AI2O3 or SiO2 Y Y O- • AI2O3 or SiO2 X Y Y X O- X O- X O- Y Y Y Y Y Mobile Liquid phase with solute X and Y Y Y Y
  • Chromatography Techniques Column Chromatography Thin Layer Chromatography Adsorption chromatography Adsorption chromatography• Solute adsorb on stationary solid phase • Solute adsorb on stationary solid phase Mobile Liquid phase (solvent) with solute X and Y X Stationary phase solid O- Stationary phase solid X X AI2O3 or SiO2 Y Y O- • AI2O3 or SiO2 X Y Y X O- X O- X O- Y Y Y Y Y Mobile Liquid phase with solute X and Y Stationary phase - solid silica SiO2 or alumina AI2O3 Y Y Y Mobile phase – liquid solvent move by capillary action Mobile phase – liquid solvent added from top (Eluant) Adsorption of solute on stationary solid phase Solute in mobile phase adsorb on solid X - Adsorb strongly – travel slowly X - Strongly adsorb – travel slowly, eluted last Y – Adsorb weakly – travel fast with solvent Y - Weakly adsorb – travel fast with solvent, eluted first
  • Chromatography Techniques Column Chromatography Thin Layer Chromatography Adsorption chromatography Adsorption chromatography• Solute adsorb on stationary solid phase • Solute adsorb on stationary solid phase Mobile Liquid phase (solvent) with solute X and Y X Stationary phase solid O- Stationary phase solid X X AI2O3 or SiO2 Y Y O- • AI2O3 or SiO2 X Y Y X O- X O- X O- Y Y Y Y Y Mobile Liquid phase with solute X and Y Stationary phase - solid silica SiO2 or alumina AI2O3 Y Y Y Mobile phase – liquid solvent move by capillary action Mobile phase – liquid solvent added from top (Eluant) Adsorption of solute on stationary solid phase Solute in mobile phase adsorb on solid X - Adsorb strongly – travel slowly X - Strongly adsorb – travel slowly, eluted last Y – Adsorb weakly – travel fast with solvent Y - Weakly adsorb – travel fast with solvent, eluted first
  • Chromatography Techniques Components separated by Paper/TLC can be identified using Rf value • Rf = Retention factor for a given eluent. • Measured the distance from original spot to the centre of a particular component to the solvent front Rf value for green spot Rf value for blue spot = 3cm/12cm = 6cm/12cm = 0.25 = 0.5http://home.earthlink.net/~dayvdanls/photolab/photolab7.htm
  • Chromatography Techniques Components separated by Paper/TLC can be identified using Rf value • Rf = Retention factor for a given eluent. • Measured the distance from original spot to the centre of a particular component to the solvent front Rf value for green spot Rf value for blue spot = 3cm/12cm = 6cm/12cm = 0.25 = 0.5 Paper Chromatography Uses of Paper/TLC/Column chromatography •Separate dyes in food colourings/plant pigmentshttp://home.earthlink.net/~dayvdanls/photolab/photolab7.htm
  • Chromatography Techniques Components separated by Paper/TLC can be identified using Rf value • Rf = Retention factor for a given eluent. • Measured the distance from original spot to the centre of a particular component to the solvent front Rf value for green spot Rf value for blue spot = 3cm/12cm = 6cm/12cm = 0.25 = 0.5 Paper Chromatography Uses of Paper/TLC/Column chromatography Column Chromatography •Separate dyes in food colourings/plant pigments • Separation/Preparative/Collection of pigmentshttp://home.earthlink.net/~dayvdanls/photolab/photolab7.htm
  • Chromatography Techniques Components separated by Paper/TLC can be identified using Rf value • Rf = Retention factor for a given eluent. • Measured the distance from original spot to the centre of a particular component to the solvent front Rf value for green spot Rf value for blue spot = 3cm/12cm = 6cm/12cm = 0.25 = 0.5 Paper Chromatography Uses of Paper/TLC/Column chromatography Column Chromatography •Separate dyes in food colourings/plant pigments • Separation/Preparative/Collection of pigments • Detection of amino acids in mixture TLC Chromatography For colour formation of amino acids • Spray with ninhydrin for colour formation • Shine UV light for fluorescent compounds to formhttp://home.earthlink.net/~dayvdanls/photolab/photolab7.htm
  • Video on Chromatographic Separation Click here to view TLC Click here to view TLCClick here on column chromatography Click here to view amino acid separation
  • AcknowledgementsThanks to source of pictures and video used in this presentationThanks to Creative Commons for excellent contribution on licenseshttp://creativecommons.org/licenses/Prepared by Lawrence KokCheck out more video tutorials from my site and hope you enjoy this tutorialhttp://lawrencekok.blogspot.com