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Materials
- Mortar & pestle
- Extraction solvent
- Hot plate
- Isopropyl alcohol
- Water
- Chromatography
solvent
- Chromatography
paper or TLC strips
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 Paper or Thin-layer Chromatography
- Remove chlorophyll
and other pigments: a. Dip each tissue in boiling water for 30
sec to kill the protoplasm in the cells. b. Put each tissue in
a beaker of alcohol and set the beaker on a hot plate until the tissue
is colorless (approx. 30 sec). Be careful, alcohol is flammable.
- Prepare plant tissue
extracts by grinding the appropriate tissue in a few milliliters of
solvent in a mortar and pestle. Possible solvents are sterile water;
methyl alcohol, ethyl alcohol, or acetone.
- Use paper chromatography
or TLC to separate the extract into fractions. (Possible solvents: 1
part 2% NaOH: 2 parts isopropyl alcohol; or 9 Petroleum ether: 1 acetone.
- Use a hole punch
to cut disks from the chromatogram and perform the disk
diffusion assay to determine which fraction contains the active
ingredients.
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