Thin layer chromatography (TLC) is a technique used to separate complex mixtures between two phases, a stationary phase and a mobile phase. In this document, TLC is used to separate photosynthetic pigments in leaves and anthocyanins in plant tissues. The method involves grinding leaf tissue, applying the extract to a TLC plate, developing the plate in a solvent, and observing the separated pigments. Different pigments are identified by their distinct colors and retention factor values. TLC is shown to effectively separate the various pigments present in coleus leaves.
Thin layer chromatography (TLC) is a technique used to separate complex mixtures between two phases, a stationary phase and a mobile phase. In this document, TLC is used to separate photosynthetic pigments in leaves and anthocyanins in plant tissues. The method involves grinding leaf tissue, applying the extract to a TLC plate, developing the plate in a solvent, and observing the separated pigments. Different pigments are identified by their distinct colors and retention factor values. TLC is shown to effectively separate the various pigments present in coleus leaves.
Thin layer chromatography (TLC) is a technique used to separate complex mixtures between two phases, a stationary phase and a mobile phase. In this document, TLC is used to separate photosynthetic pigments in leaves and anthocyanins in plant tissues. The method involves grinding leaf tissue, applying the extract to a TLC plate, developing the plate in a solvent, and observing the separated pigments. Different pigments are identified by their distinct colors and retention factor values. TLC is shown to effectively separate the various pigments present in coleus leaves.
Thin layer chromatography (TLC) is a technique used to separate complex mixtures between two phases, a stationary phase and a mobile phase. In this document, TLC is used to separate photosynthetic pigments in leaves and anthocyanins in plant tissues. The method involves grinding leaf tissue, applying the extract to a TLC plate, developing the plate in a solvent, and observing the separated pigments. Different pigments are identified by their distinct colors and retention factor values. TLC is shown to effectively separate the various pigments present in coleus leaves.
vegetables • Place ca. 3 cm2 leaf in mortar Method • [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 Method
• Supernatant and residue from grinding leaf in propanone 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
1.5 cm 1.0 2.0 3.0 4.0 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
1.5 cm 1.0 2.0 3.0 4.0 Method • Repeat process until a second line is produced (or one line across the whole plate)
1.5 cm 1.0 2.0 3.0 4.0 Method • Plate into a 600 cm3 beaker, replace foil
5:3:2 Pigment Colour RF value carotene yellow-orange 0.91 pheophytin a grey 0.75 pheophytin b light grey 0.63-0.75 chlorophyll a blue green 0.63 chlorophyll b green 0.58 xanthophylls yellow 0.53 xanthophylls yellow 0.47 xanthophylls yellow 0.32 Coleus Anthocyanin results
