The Secret of Color Separation

Introduction

Colors can be separated by chromatography. Scientists use chromatography method to separate

molecules of different masses. In coloring materials, molecules with different masses may have
different colors.

Plants have various separable pigments to generate beautiful colors on its leaves and flowers.
They may have carotene (orange), xanthophyl (yellow), chlorophyl (green), and anthoxianine
(red) pigments. The pigments can be separated by the chromatography. This method needs a
suitable paper as separation medium and a liquid to dissolve the pigments. One of examples of
pigment that is usually used in our daily life is food coloring.

A quantitative indication of how far a particular compound travels in a particular solvent is

known as retention factor (Rf). The retention factor is given by:

distance the color traveled, measured from center of the color band to the point where the
color was placed

total distance traveled by solvent

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You are asked to design an experiment to determine the component of ink of marker, food
coloring, and leaves and then calculate their Rf values.

In the following list, there are some materials you may need in this experiment:

1. Alcohol 70 % 1 bottle
2. Water 1 bottle
3. Plastic box 2
4. Paper clips 10 pieces
5. Experiment guideline 1 plastic bag
(scrap paper and green marker)
6. Different type of papers 4 types :
- rough filter paper
- smooth filter paper
- scrap paper
- copy/ HVS paper
7. Spinach leaves (green & red)
8. Food coloring 3 (yellow, green, and red)
9. Ruler 1 piece
10. Watch 1 piece
11. Medium plastic bag 10 pieces
12. Labels 1 packet
13. Pallet 1 piece
14. Towel 1 piece
15. Water bucket 1 piece
16. Tooth pick 1 packet
17. Coin 2
18. Pipette 3

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 19. Supporting aluminum bar 2

Experiment 1

1. Open the experiment guideline.

2. Make a dot by using a marker on one piece of scrap paper located 2 cm from the
bottom.
3. Prepare a plastic box and fill with water.
4. Hang the paper. Immerse the bottom (approximately 1 cm) of paper in the water. After
about 5 minutes take the paper and observe it. Record your observation.
(score 6)

Experiment 2

1. Design an experiment to show the separation differences among four types of papers by
drawing your experimental set up on the answer sheet.
2. Do the experiment using your experimental design. Hang the paper and suspend it.
3. After about 10 minutes, take the papers and observe it. Record your observation result.
(score 15)

Based on your experiment, which paper has the best performance to separate the colors?
Explain your answer. (score 6)

4. After the experiment is finished, place the papers into the plastic bag. Both paper and
plastic bag should be labeled (before you place the paper make sure that the paper is
dry ).
(Score 6)

Experiment 3

Always use the paper that gives the best result in previous experiment

1. Use a set of food coloring. Choose primary colors and make one secondary color, and
make one tertiary color. Write down what colors you used and what are the results in
table form on the answer sheet (Make the tertiary color by mixing one secondary color
and one primary color).
(Score 6)

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2. Observe the chromatography process for one primary, one secondary and one tertiary
color until the colors movement stops (±10 minutes). Dry the paper approximately 10
minutes.

3. Based on your results, write the color appearances of the band and calculate the Rf
values from each band. (before you calculate the Rf value make sure that the paper is
dry )
(Score 15)

4. Answer the questions below:

Analyze your Rf data. Are there any significant differences between Rf values of the same
color from different bands?

(Score 1)

5. After finishing the experiment, place the papers into plastic bag and give label to each
paper (before you place the papers make sure that they are dry).
(Score 4)

Experiment 4

1. Design an experiment to show that the chromatography results are influenced by the
type of the solvents.
2. Draw your design on the answer sheet and do the experiment. Materials used for this
experiment are the red and green spinaches.
3. Scrape the surface of their leaves of spinaches to release the pigments from the
spinaches. Use a coin if necessary. In this experiment you have to identify each pigment
found in the spinaches. It needs approximately 15 minutes to get the results.
4. Calculate their Rf values. Don’t forget to record your results on the answer sheet.
(Score 6)
5. After finishing the experiment place the paper results into plastic bag and give the
correct label (before you place the paper make sure that the paper is dry )
(Score 4 )
6. Based on your experiment, answer the following questions :
a. Which solvent is better for chromatography of leaves?
(Score 1)
b. Is the order of the pigments similar for different leaves?
(Score 1)
c. Write the pigment order for each leaf (from the bottom of paper that immersed in the
solvent).
(Score 4)

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 d. Do the same color originating from different leaves have the same Rf values? Write
the Rf value for each pigment.
(Score 4)

Write all the conclusions based on all of your experimental results.

(Score 6)

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