IB Chemistry on Paper Chromatography, Thin Layer and Column Chromatography

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

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