This document discusses the extraction and separation of chlorophyll, carotenoids, and xanthophylls from plant leaves using thin-layer chromatography. It provides background on the technique, including that leaf extracts are placed as spots on a plate coated with silica. Mobile solvents like acetone and alcohol are used to separate the pigments based on their retention values. The document also gives the historical context of thin-layer chromatography and some of the key plant pigments that can be separated this way such as chlorophyll a, chlorophyll b, lutein, and beta-carotene.

1. The extraction and separation of
chlorophyll (α+β) carotin and
xanthophyll in fresh green leaves
Eugene Ong | eugene@usm.my
Institute for Research in Molecular Medicine
Universiti Sains Malaysia
27 June 2014
SMK Penang Free

2. Plant Physiol. Apr 1928; 3(2): 211–216.

3. Plant Physiol. Apr 1928; 3(2): 211–216.

5. Thin-Layer Chromatography
lutein (yellow)
chlorophyll a (yellow – green)
chlorophyll b (dark green)
β-carotene (yellow)
pheophytin (brown – green)
xanthophylls

6. STATIONARY PHASE
Plate coated with silica.
LEAF EXTRACT
MOBILE PHASE
Solvents (acetone,
alcohol, hexane, etc)
xy
Retention value,
Rf = x
y
Thin-Layer Chromatography

7. STATIONARY PHASE
Plate coated with silica.
LEAF EXTRACT
MOBILE PHASE
Solvents (acetone,
alcohol, hexane, etc)
1938, Ukraine
Acetone, 1610, France
Chlorophyll, 1817, France
Thin-Layer Chromatography
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