Chromatography of Chlorophyll

Paper chromatography is a useful way to separate out the different pigment

molecules in chlorophyll. Using a solvent, pigments found in plants are
separated. The pigments are able to dissolve in the solvent and separate
from each other. When they separate, the different colors of the pigments
will be visible. This way we can tell which pigments are inside the leaves.
The pigments will be absorbed along a paper column until they reach the top.
Larger pigment molecules will move up the paper slower and , smaller
pigment molecules will move up the paper faster.

Procedure:

1. Obtain a strip of chromatography paper.

2. Measure 1 inch from the bottom and mark a pencil mark ( MUST BE PENCIL)
3. Obtain a piece of a leaf wider than the strip of paper
4. Using a coin, roll the coin across the pencil mark smooshing the
chlorophyll from the leaf on the pencil line.
5. Now, the line should be heavily covered in green chlorophyll.
6. Very carefully place the strip into the jar containing the solvent.
7. Be careful not to get the line in the liquid solvent!
8. Cover the jar. Let the jar sit undisturbed until the liquid has
moved up to very near the top of the strip of paper.
(DON’T let the liquid run off the strip!)

9. Attach the strip to the paper here:

Label the different color pigments as;

COLOR REFLECTED PIGMENT NAME
Dark green to blueish-green chlorophyll a
Yellowish green to (olive) chlorophyll b
Orange- yellow carotene
Lemon yellow xanthophyll

Answer the questions:

1. Which pigment moved the farthest up the paper? ____________________
2. Which pigment moved the shortest distance up the paper? _______________
3. If the largest molecule moved the slowest and the smallest molecule moved
the fastest up the paper, then name the molecules in order from largest to
smallest Largest _____________________________
______________________________
______________________________
Smallest ______________________________

4. In the autumn, when the leaves change color, what pigments become
weaker in expression? ______________________ and
_________________________
5. In the autumn, which pigments become more DOMINANT?
__________________________________________________