General Biology 1

Science, Technology, Engineering and Mathematics

BLOCK NAMES GROUP #

LABORATORY SCHEDULE DATE

Laboratory Activity 7- A
Paper Chromatography of Leaf Pigments

INTRODUCTION

Paper chromatography is a process that uses special filter paper to separate and identify the different
substances in a mixture. Chromatography means “to write with color”. The substances in the mixture dissolve in
the alcohol and move up the paper. The heavier substances move up the paper more slowly. The lighter
substances move up the paper more quickly. So, heavy and light substances get separated from one another on the
paper. Plants contain chlorophyll, a green pigment, as well as carotenoid, a pigment that ranges in color from red
to orange to yellow.

MATERIALS
• 2 different types of leaves ruler
• graduated cylinder scissors
• Filter paper/ chromatography paper strip pencil/ colored pencils
• Ethanol / Isopropyl alcohol penny

PROCEDURES
1. Draw a line in pencil across the paper 1.5 cm from the bottom.
2. Starting at the line, taper the bottom of the paper with scissors.
3. Your chromatogram should resemble the picture shown below (Figure 1).
4. Place leaf over the line on the filter strip.
5. Using a penny, rub the leaf to transfer pigment onto the line.
6. Move the leaf and repeat several times until a dark green line of leaf pigment is visible on the pencil line.
7. Place the filter strip into the graduated cylinder with 10 ml of ethanol/ isopropyl alcohol.
(DO NOT INHALE THE SOLVENT).
8. Fold the top of the paper over the edge of the graduated cylinder. Be sure the paper doesn’t sink further
into the solvent. The green line should not be in the solvent.
9. Wait for fifteen minutes, or until the pigments fully separate. There should be different colors of pigment
visible on the strip.
10. Remove the filter strip and let it dry.
11. Sketch your pigment bands.

Place leaf over pencil line and rub

Figure 1.

Paper Chromatography of Leaf Pigments

DATA AND OBSERVATIONS

Type of Leaf Color of Pigment Name of Pigment

Identifying the pigments: Use the information below to identify which pigments are in your leaf.
Chlorophyll a- bright green yellow
Chlorophyll b- dull blue green or khaki
Carotene- orange or yellow
Xanthophyll- pale yellow

Results:

Filter Paper Use colored pencils or crayons to draw your

observations

Before paper chromatography

After paper chromatography

QUESTIONS FOR RESEARCH

1. Does your data support this statement: “Chlorophyll is the most abundant and important photosynthetic plant
pigment”? Explain.

2. Why is paper chromatography an appropriate technique to use to determine if different pigments are present in
leaf?

Paper Chromatography of Leaf Pigments

3. Why do most leaves contain one or more than one type of pigment? What is the purpose of pigments?

4. Explain the technique of paper chromatography.

5. Based on what you have learned, explain why leaves tend to change color in the fall.

6. Leaves in New England change color in the fall. However, leaves in Florida do not change color in the fall.
Why is this so? (Hint: Think of the difference between the two locations that might act as a trigger for leaves to
change color in the fall).

REFERENCES:

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Paper Chromatography of Leaf Pigments