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AP Chemistry Lab

Paper Chromatography

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 all
involve a stationary phase an a mobile phase. If each component has a different affinity for the mobile and
stationary phase, they can be separated. In paper chromatography the stationary phase is paper and the
mobile phase is the solvent. Capillary action draws the mobile phase up the paper. If the component has a
strong attraction for the mobile phase, it tends to move with it. If the component has a strong affinity for
the paper, it stays behind. When the mobile phase is water, the more polar components travel the furthest.
Rf values compare the distance the solute traveled compared to the solvent.
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. Finally you will be able to determine the affinity each color has for the mobile and
stationary phases.
Materials:
1 piece of chromatography paper
1 large beaker
6 Toothpicks
Test tube rack

3 FD&C food dyes


1 watch glass that fits over the beaker
Ruler
3 different flavors of Kool-aid

Method:
1. Wash your hands with soap and water and dry thoroughly. Handle the paper by the edges as
much as possible as skin oils and moisture can adversely affect the experiment.
2. Draw a light pencil (NOT PEN) line across the strip 2 cm from the bottom. At the top mark in
pencil which of the dyes or Kool-aid will be used on the strip.
3. 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. Place a dot of each of the 3 FD&C dyes along with each of the Kool-aid flavors 1-2 cm apart
from each other (see figure 1).

4. Allow the spots to dry and repeat the procedure 3-4 times. The spots should be fairly small and
concentrated when you finish.
5. Put a small amount of distilled water into the bottom of the beaker. The depth should only be about
1 cm. It must be less than 2 cm.
6. Put the piece of paper in the beaker. Make sure the dye spot remains above the water level.
7. To keep the paper from falling in the solvent, place a stirring rod across the top of the beaker and
secure the paper by taping it to the stirring rod.
8. Cover the beaker with a watch glass. This keeps air currents out of the developing chamber.
9. Allow the paper to develop until the solvent front nears the top of the paper. (about 20 minutes)
10. When the solvent front has nearly reached the top, remove the paper from the chamber and mark the
solvent front with a pencil.
11. Place the chromatography paper on a paper towel and allow it to dry.
12. 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.
13. 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. If the solution was a mixture, be sure to
record each dye observed.
14. 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
Determine the Rf value for each of the dyes. If the solution you used was a mixture, be sure to calculate the
Rf value for each dye in the solution.