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IB Chemistry on Paper Chromatography, Thin Layer and Column Chromatography
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http://lawrencekok.blogspot.com
Prepared by Lawrence Kok
Tutorial on Chromatographic Techniques, Paper Chromatography , Thin Layer and Column Chromatography.
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
Chromatography Techniques
Separation analysis
Adsorption Chromatography
Chromatography
Partition Chromatography
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
Chromatography Techniques
Separation analysis
Adsorption Chromatography
Chromatography
Partition Chromatography
Partition Chromatography• Components distribute bet TWO immisible liquid phase • Depends on relative solubility bet TWO phase• Solutes bonds to stationary phase or mobile phase
Adsorption Chromatography• Component adsorb on solid stationary phase • Depends on polarity of stationary phase, mobile phase and solutes• 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• 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
Chromatography Techniques
Separation analysis
Paper Chromatography
Thin Layer Chromatography
Adsorption Chromatography
Chromatography
Partition Chromatography
Column Chromatography
Partition Chromatography• Components distribute bet TWO immisible liquid phase • Depends on relative solubility bet TWO phase• Solutes bonds to stationary phase or mobile phase
Adsorption Chromatography• Component adsorb on solid stationary phase • Depends on polarity of stationary phase, mobile phase and solutes• 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
Application• Detection of amino acids in a mixture• Different dyes in food colouring• Separation of plant pigments
Y adsorb strongly
Application• Detection of amino acids in a mixture• Different dyes in food colouring• Separation of plant pigments
Application• Preparative/collection of sample of pigments
X adsorb strongly
Chromatography Techniques
Chromatography
Partition Chromatography• components distribute between 2 immisible liquid phase • relative solubility in 2 phase• bonds strongly to mobile phase – move faster
Adsorption Chromatography• components adsorp on solid stationary phase
Stationary phase has a layer of liquid
Stationary Liquid phase
Stationary phase • solid• AI2O3
• SiO2
O-O-O-O-
O-
Chromatography Techniques
Chromatography
Partition Chromatography• components distribute between 2 immisible liquid phase • relative solubility in 2 phase• bonds strongly to mobile phase – move faster
Adsorption Chromatography• components adsorp on solid stationary phase
Stationary phase has a layer of liquid
Mobile liquid phase containing X and Y X
X
X
Y
Y
Y
Stationary Liquid phase
Stationary phase • solid• AI2O3
• SiO2
O-O-O-O-
O-
Y
Y
YY
Mobile liquid phase containing X and Y
X
X
X
Chromatography Techniques
Chromatography
Partition Chromatography• components distribute between 2 immisible liquid phase • relative solubility in 2 phase• bonds strongly to mobile phase – move faster
Adsorption Chromatography• components adsorp on solid stationary phase
Stationary phase has a layer of liquid
Mobile liquid phase containing X and Y X
X
X
Y
Y
Y
Stationary Liquid phase
Stationary phase has a layer of liquid
Y
YY
Y
XXX
XX
X Mobile Iiquid phase containing X
Stationary phase • solid• AI2O3
• SiO2
O-O-O-O-
O-
Y
Y
YY
Mobile liquid phase containing X and Y
X
X
X
O-O-O-
YY
YY
X XXX
Mobile liquid phase containing X
Y adsorb strongly
Chromatography Techniques
Chromatography
Partition Chromatography• components distribute between 2 immisible liquid phase • relative solubility in 2 phase• bonds strongly to mobile phase – move faster
Adsorption Chromatography• components adsorp on solid stationary phase
Stationary phase has a layer of liquid
Mobile liquid phase containing X and Y X
X
X
Y
Y
Y
Stationary Liquid phase
Stationary phase has a layer of liquid
Y
YY
Y
XXX
XX
X Mobile Iiquid phase containing X
Stationary phase • solid• AI2O3
• SiO2
O-O-O-O-
O-
Y
Y
YY
Mobile liquid phase containing X and Y
X
X
X
O-O-O-
YY
YY
X XXX
Mobile liquid phase containing X
Y adsorb strongly
XX
X
X
YYY
Y
Separation of X and Y
X XX
YYY
Chromatography Techniques
Chromatography
Partition Chromatography• components distribute between 2 immisible liquid phase • relative solubility in 2 phase• bonds strongly to mobile phase – move faster
Adsorption Chromatography• components adsorp on solid stationary phase
Stationary phase has a layer of liquid
Mobile liquid phase containing X and Y X
X
X
Y
Y
Y
Stationary Liquid phase
Partition –distribution of solute X and Y between 2 liquid phase• X more soluble in mobile phase (move with mobile liquid phase)• Y less soluble in mobile phase (stay on stationary liquid phase)
Stationary phase has a layer of liquid
Y
YY
Y
XXX
XX
X Mobile Iiquid phase containing X
Stationary phase • solid• AI2O3
• SiO2
O-O-O-O-
O-
Adsorption– solute X and Y adsorb temporary on the solid• Y adsorb strongly on solid phase, eluted slower• X in liquid mobile phase, eluted faster
Y
Y
YY
Mobile liquid phase containing X and Y
X
X
X
O-O-O-
YY
YY
X XXX
Mobile liquid phase containing X
Y adsorb strongly
XX
X
X
YYY
Y
Separation of X and Y
X XX
YYY
Chromatography Techniques
Paper Chromatography
Partition chromatography• Distribution solute bet both liquid phase• Depends on relative solubility
Thin Layer Chromatography
Adsorption chromatography• Solute adsorb on stationary solid phase
Stationary phase - Cellulose paper • absorb water on its surface
Mobile Liquid phase with solute X and Y
Y
Y
Y
X
X
Stationary phase solid• AI2O3 or SiO2
O-O-O-
O-O-
Mobile Liquid phase with solute X and Y
Y
Y
Y
X
X
Chromatography Techniques
Paper Chromatography
Partition chromatography• Distribution solute bet both liquid phase• Depends on relative solubility
Aqueous liquid phase on surface of stationary phase (paper)
Mobile liquid phase - solvent
Solvent move by capillary action
Thin Layer Chromatography
Adsorption chromatography• Solute adsorb on stationary solid phase
Stationary phase - solid silica SiO2 or alumina AI2O3
Separation depends on how strongly solute adsorb on solid
Mobile phase – liquid solvent move by capillary action
Adsorption of solute on stationary solid phase
Stationary phase - Cellulose paper • absorb water on its surface
Mobile Liquid phase with solute X and Y
Y
Y
Y
X
X
Stationary phase solid• AI2O3 or SiO2
O-O-O-
O-O-
Mobile Liquid phase with solute X and Y
Y
Y
Y
X
X
Chromatography Techniques
Paper Chromatography
Partition chromatography• Distribution solute bet both liquid phase• Depends on relative solubility
Aqueous liquid phase on surface of stationary phase (paper)
Mobile liquid phase - solvent
Solvent move by capillary action
Thin Layer Chromatography
Adsorption chromatography• Solute adsorb on stationary solid phase
Stationary phase - solid silica SiO2 or alumina AI2O3
Separation depends on how strongly solute adsorb on solid
Mobile phase – liquid solvent move by capillary action
Adsorption of solute on stationary solid phase
Stationary phase - Cellulose paper • absorb water on its surface
Mobile Liquid phase with solute X and Y
Y
Y
Y
X
X
Stationary phase solid• AI2O3 or SiO2
O-O-O-
O-O-
Mobile Liquid phase with solute X and Y
Y
Y
Y
X
X
Chromatography Techniques
Thin Layer Chromatography
Adsorption chromatography• Solute adsorb on stationary solid phase
Mobile Liquid phase (solvent) with solute X and Y
YY Y
X
X Stationary phase solid• AI2O3 or SiO2
O-O-O-
O-O-
Mobile Liquid phase with solute X and Y
YY
Y
X
X
Column Chromatography
Adsorption chromatography• Solute adsorb on stationary solid phase
X
X
Y
Y
Stationary phase solid AI2O3 or SiO2
Y YY
X
Y
Chromatography Techniques
Solute in mobile phase adsorb on solid
Mobile phase – liquid solvent added from top (Eluant)
X - Strongly adsorb – travel slowly, eluted lastY - Weakly adsorb – travel fast with solvent, eluted first
Thin Layer Chromatography
Adsorption chromatography• Solute adsorb on stationary solid phase
Adsorption of solute on stationary solid phase
Mobile Liquid phase (solvent) with solute X and Y
YY Y
X
X Stationary phase solid• AI2O3 or SiO2
O-O-O-
O-O-
Mobile Liquid phase with solute X and Y
YY
Y
X
X
Column Chromatography
Adsorption chromatography• Solute adsorb on stationary solid phase
X
X
Y
Y
Stationary phase solid AI2O3 or SiO2
Y YY
X - Adsorb strongly – travel slowlyY – Adsorb weakly – travel fast with solvent
X
Y
Stationary phase - solid silica SiO2 or alumina AI2O3
Mobile phase – liquid solvent move by capillary action
Chromatography Techniques
Solute in mobile phase adsorb on solid
Mobile phase – liquid solvent added from top (Eluant)
X - Strongly adsorb – travel slowly, eluted lastY - Weakly adsorb – travel fast with solvent, eluted first
Thin Layer Chromatography
Adsorption chromatography• Solute adsorb on stationary solid phase
Adsorption of solute on stationary solid phase
Mobile Liquid phase (solvent) with solute X and Y
YY Y
X
X Stationary phase solid• AI2O3 or SiO2
O-O-O-
O-O-
Mobile Liquid phase with solute X and Y
YY
Y
X
X
Column Chromatography
Adsorption chromatography• Solute adsorb on stationary solid phase
X
X
Y
Y
Stationary phase solid AI2O3 or SiO2
Y YY
X - Adsorb strongly – travel slowlyY – Adsorb weakly – travel fast with solvent
X
Y
Stationary phase - solid silica SiO2 or alumina AI2O3
Mobile phase – liquid solvent move by capillary action
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
http://home.earthlink.net/~dayvdanls/photolab/photolab7.htm
Chromatography Techniques
Rf value for green spot= 3cm/12cm= 0.25
Rf value for blue spot= 6cm/12cm= 0.5
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
http://home.earthlink.net/~dayvdanls/photolab/photolab7.htm
Uses of Paper/TLC/Column chromatography• Separate dyes in food colourings/plant pigments
Chromatography Techniques
Rf value for green spot= 3cm/12cm= 0.25
Rf value for blue spot= 6cm/12cm= 0.5
Paper Chromatography
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
http://home.earthlink.net/~dayvdanls/photolab/photolab7.htm
Uses of Paper/TLC/Column chromatography• Separate dyes in food colourings/plant pigments• Separation/Preparative/Collection of pigments
Chromatography Techniques
Rf value for green spot= 3cm/12cm= 0.25
Rf value for blue spot= 6cm/12cm= 0.5
Paper Chromatography
Column Chromatography
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
http://home.earthlink.net/~dayvdanls/photolab/photolab7.htm
Uses of Paper/TLC/Column chromatography• Separate dyes in food colourings/plant pigments• Separation/Preparative/Collection of pigments• Detection of amino acids in mixture
Chromatography Techniques
Rf value for green spot= 3cm/12cm= 0.25
Rf value for blue spot= 6cm/12cm= 0.5
For colour formation of amino acids• Spray with ninhydrin for colour formation• Shine UV light for fluorescent compounds to form
Paper Chromatography
TLC Chromatography
Column Chromatography
Click here to view TLC
Click here on column chromatography
Click here to view TLC
Click here to view amino acid separation
Video on Chromatographic Separation
Acknowledgements
Thanks to source of pictures and video used in this presentation
Thanks to Creative Commons for excellent contribution on licenseshttp://creativecommons.org/licenses/
Prepared by Lawrence Kok
Check out more video tutorials from my site and hope you enjoy this tutorialhttp://lawrencekok.blogspot.com