Leaf Pigment Chromatography
Purpose: To separate and compare bands of pigment from a spinach leaf.
Hypothesis: How many different pigments do you think are present in a spinach
leaf’s cells? (Answer as an if…then statement!)
Materials: chromatography solvent, spinach, coin, chromatography paper, pencil,
tape, chromatography jars with lids, metric ruler (Write in bulleted or numbered
columns.)
Procedure:
1. Measure and cut a strip of chromatography paper to fit inside of the jar so
that the tip just touches the solvent.
2. Measure 1.5 cm from the end of the strip and draw a pencil line.
3. Place the spinach leaf on the line and use the coin to smash pigment into the
strip of chromatography paper.
4. Mark an X next to the line of spinach pigment (this is the point of origin).
5. Tape the other end of the strip to the underside of the jar lid and screw on
the lid (making sure the spinach pigment is NOT submerged in the solvent).
6. Wait and observe the pigments separate until the solvent reaches 2 cm below
the top of the strip.
7. Remove the strip and immediately mark the solvent front with a pencil line.
8. Measure the solvent front and record in your table.
9. Measure the distance from the point of origin to the top of each pigment band
and record in your table, describe the color of each band as well.
10. Divide the distance each pigment traveled by the solvent front to calculate
the Rf value. (Round to the nearest hundredth.) Record these in your chart
as well.
Results:
Solvent front: _________________ mm
Band
Color
Spinach Leaf Pigments
Distance pigment
traveled (mm)
Rf value
1
2
3
4
5
6
Analysis: (Write in complete sentences!)
1. How many bands are visible and does the data support or refute your
hypothesis?
2. Why are we unable to see all of these pigments in the leaf?
3. Why does the plant need so many different pigments?