Plant Pigment Chromatography
Introduction: Paper chromatography is a technique used to separate substances in
a mixture based on the movement of the different substances up a piece of paper
by capillary action. Pigments extracted from plant cells contain a variety of
molecules, such as chlorophylls, beta carotene, and xanthophyll, that can be
separated using paper chromatography. A small sample of plant pigment placed on
chromatography paper travels up the paper due to capillary action. Beta carotene is
carried the furthest because it is highly soluble in the solvent and because tit forms
no hydrogen bonds with the chromatography paper fibers. Xanthophyll contains
oxygen and does not travel quite as far with the solvent because it is less soluble the
beta carotene and forms hydrogen bonds with the paper. Chlorophylls are bound more
tightly to the paper than the other two, so they travel the shortest distance.
Objectives:
• Separate plant pigments in Spinach leaves
• Calculate the Rf values for the pigments
Materials
100 mL graduated cylinder Aluminum foil
Chromatography paper Pencil Spinach leaves
Scissors Coin Chromatography Solvent
Ruler
Procedure
The solvent in this experiment is flammable and poisonous. Be sure there are no open flames in the lab during
the experiment. Avoid inhaling the fumes. Close the solvent bottle immediately after you are finished pouring the
solvent. Notify your teacher is any accidents occur.
1. Pour 10 mL of chromatography solvent into a 100 mL graduated cylinder. Cover the cylinder with aluminum foil.
2. Cut the end of the chromatography paper into a point.
3. Draw a pencil line 2.0 cm above the pointed end of the paper.
4. Use the coin to extract pigments from the spinach leaf. Place a small amount of
the leaf on top of the pencil line. Use the edge of the coin to push the plant cells
into the chromatography paper. Repeat the procedure 10 times making sure to
use a different part of the leaf each time.
5. Place the chromatography paper in the cylinder so that the pointed end just
touches the solvent. Make are the pigment is not in the solvent.
6. Fold over the paper at the top of the cylinder and recover with aluminum foil.
Wait until the solvent is approximately 2 cm from the top of the paper. Remove the paper and mark the solvent front before
it evaporates.
7. Return the solvent to the stock bottles.
Introduction: Paper chromatography is a technique used to separate substances in
a mixture based on the movement of the different substances up a piece of paper
by capillary action. Pigments extracted from plant cells contain a variety of
molecules, such as chlorophylls, beta carotene, and xanthophyll, that can be
separated using paper chromatography. A small sample of plant pigment placed on
chromatography paper travels up the paper due to capillary action. Beta carotene is
carried the furthest because it is highly soluble in the solvent and because tit forms
no hydrogen bonds with the chromatography paper fibers. Xanthophyll contains
oxygen and does not travel quite as far with the solvent because it is less soluble the
beta carotene and forms hydrogen bonds with the paper. Chlorophylls are bound more
tightly to the paper than the other two, so they travel the shortest distance.
Objectives:
• Separate plant pigments in Spinach leaves
• Calculate the Rf values for the pigments
Materials
100 mL graduated cylinder Aluminum foil
Chromatography paper Pencil Spinach leaves
Scissors Coin Chromatography Solvent
Ruler
Procedure
The solvent in this experiment is flammable and poisonous. Be sure there are no open flames in the lab during
the experiment. Avoid inhaling the fumes. Close the solvent bottle immediately after you are finished pouring the
solvent. Notify your teacher is any accidents occur.
1. Pour 10 mL of chromatography solvent into a 100 mL graduated cylinder. Cover the cylinder with aluminum foil.
2. Cut the end of the chromatography paper into a point.
3. Draw a pencil line 2.0 cm above the pointed end of the paper.
4. Use the coin to extract pigments from the spinach leaf. Place a small amount of
the leaf on top of the pencil line. Use the edge of the coin to push the plant cells
into the chromatography paper. Repeat the procedure 10 times making sure to
use a different part of the leaf each time.
5. Place the chromatography paper in the cylinder so that the pointed end just
touches the solvent. Make are the pigment is not in the solvent.
6. Fold over the paper at the top of the cylinder and recover with aluminum foil.
Wait until the solvent is approximately 2 cm from the top of the paper. Remove the paper and mark the solvent front before
it evaporates.
7. Return the solvent to the stock bottles.
