AP Chemistry Lab

Paper Chromatography

Please do not write on this paper. Return to the instructor when finished with the lab.
Introduction: Chromatography is a means of separating mixtures of substances that are in the same phase.
There are many types of chromatography including gas, high pressure liquid, thin film and paper. The
basic principle involved is the fact that when forced to travel some distance through for example a matrix
of paper fibers, the larger, heavier molecules travel more slowly than the lighter ones and are thus separated
from each other. It is a very common technique for chemical analysis.
Purpose: In this lab, you will perform paper chromatography on a number of food colorings. These dyes
are used in a great many consumer food products. You are to find out which dyes contain a single color,
which are mixtures, and how many different individual dyes are present in the samples. You are also to
identify the individual dye components of any mixtures, using color and Rf values to support your
determinations.
Materials:
3 pieces of chromatography paper
6 Glass test tubes
3 Toothpicks
Test tube rack

6 FD&C food dyes

Scissors
6 Al foil squares to fit top of test tubePetri dish
10 mL graduated cylinder
Ruler

Method:
1. Hypothesize about which of the available food colorings you think are single dyes and which are
mixtures. If you think the coloring is a mixture, tell which basic colors you think are in the mixture.
2. Wash your hands with soap and water and dry thoroughly. Handle the papers by the edges as
much as possible as skin oils and moisture can adversely affect the experiment.
3. Obtain 3 precut strips of filter paper. Cut the strips in half lengthwise. You should have 6 long,
thin strips when you are done. Try to remove the curl from the paper strips if you can.
4. Draw a light pencil (NOT PEN) line across the strip about 2 cm from the bottom. At the top of
each strip mark in pencil which of the 6 dyes will be used on the strip.
5. Obtain a Petri dish and place 1 drop of each dye in the dish.
6. Using one end of a toothpick, dip the wood in to the dye and use it to put a spot of dye on the pencil
mark in the center of the appropriate chromatography strip.
7. Allow the spot to dry and repeat the procedure 3-4 times. The spot should be fairly small and
concentrated when you finish.
Pencil line
x
Label

2 cm

Dye spot

8. Put about 3-4 mL of distilled water into the bottom of each of the 6 test tubes.
9. Put 1 strip of paper in each test tube. Dip the end of the strips with the dye spots into the water.
Make sure the dye spot remains above the water level.
10. Bend the top of the paper strip over the lip of the test tube and cover the top of the tube with a
small square of Al foil. The foil keeps the paper strips in place and keeps air currents out of the
developing chamber.
11. Allow the strips to develop for 30 minutes or until the solvent front nears the top of the paper.
12. When the solvent front has nearly reached the top, remove the strips from the chamber and mark the
solvent front with a pencil.
13. Place the strips on a paper towel and allow them to dry.

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14. Using a ruler, measure the distance that the water solvent traveled from the line at the bottom of the
strip to the highest point reached by the solvent front, also marked in pencil near the top of the strip.
This distance will be labeled as Dsol.
15. Measure how far each dye in the sample moved from the pencil line at the bottom of the strip. If
the dye color is a smear, use your judgment as to which part of the colored area you will measure.
BE CONSISTENT. This distance will be labeled as Ddye.
16. Clean the Petri dish. The dyes are sink safe and may be washed down the drain. Save the strips to
include in your report.
Calculations:
An important value calculated in chromatography experiments is Rf. It is a ratio of how far the substance in
the mixture traveled versus how far the solvent traveled. It is easily calculated as:
Rf = Ddye/Dsol
Rf value is inversely proportional to the molecular mass. Larger molecules have smaller Rf values and vice
versa.
Show at least one sample calculation of Rf for one of the dyes.
Report the Rf values for each dye measured in the data/calculations table.
Questions for Analysis
1. How many different single/individual dyes do there appear to be?
2. What colors are they?
3. What are the average Rf values for each of these individual colors?
4. Rank the different dye molecules (colors) from lightest to heaviest.
5. Which food colorings are mixtures?
6. Did the chromatography process separate the dye mixtures adequately?
7. Compare the Rf values of the dyes in the mixtures (along with the colors) and identify the
components of each mixture.
8. Ask the instructor for a list of which dyes are present in the food colorings. Research the molecular
weights of these different molecules and tell if they correspond relatively to the Rf values from your
experiment.
Conclusion:
Write a brief conclusion telling
1. what you conclude following your analysis.
2. if your hypotheses were accepted or rejected.
3. what you learned from this lab.
4. what the sources of error might have been.

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