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Thin Layer Chromatography Investigations Leighton Dann and Paul Beaumont Science & Plants for Schools www.saps.org.uk.

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Presentation on theme: "Thin Layer Chromatography Investigations Leighton Dann and Paul Beaumont Science & Plants for Schools www.saps.org.uk."— Presentation transcript:

1 Thin Layer Chromatography Investigations Leighton Dann and Paul Beaumont Science & Plants for Schools

2 TLC One of a number of types of chromatography: gas/liquid affinity paper

3 TLC Separation of complex mixtures between two phases: -a stationary phase -and a mobile phase

4 TLC Phases the stationary phase is a thin layer of absorbent particles, in this case silica gel the mobile phase is the solvent

5 Fisher Scientific UK. Tel: Cat No: TLC X

6 Thin Layer Chromatography Separation of: photosynthetic pigments in leaves anthocyanins in leaves, flowers, fruit and vegetables

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9 Method Place ca. 3 cm 2 leaf in mortar [If tissue is coarse, silver sand may be added] Grind until all tissue broken up Add ca. 750  l propanone Pour off supernatant into a container and seal

10 Method Supernatant and residue from grinding leaf in propanone

11 Method Mark (with a pencil) a plate 1.5 cm up from the base on the edge of both sides Use a ruler across these marks to mark two spots 2 cm apart in the middle of the plate

12 Method Use Pasteur pipette to place small drops of supernatant Dry off before adding another lot of drops Continue until there is a concentrated line of colour

13 Method Repeat process until a second line is produced (or one line across the whole plate)

14 Method Plate into a 600 cm 3 beaker, replace foil Remove plate when solvent nearly at top Immediately mark solvent front

15 Solvent system Cyclohexane : propanone : petroleum ether (low boiling point) 5 : 3 : 2

16 PigmentColourR F value caroteneyellow-orange0.91 pheophytin agrey0.75 pheophytin blight grey chlorophyll ablue green0.63 chlorophyll bgreen0.58 xanthophyllsyellow0.53 xanthophyllsyellow0.47 xanthophyllsyellow0.32

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19 Coleus

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22 Anthocyanin results


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