CHEM2410

Food Colors, Chromatography, and Flourescence

Introduction
There are hundreds of chromatography methods that are sorted into a multitude of types, but they all involve one
very simple model:
Compounds start at one end of the stationary phase
The mobile phase carries them toward the other end of the stationary phase
Some compounds move more readily than others and thus compounds can be separated
The stationary phase might be a solid or liquid and one of thousands of molecule types. It might be coated onto a
sheet or the inside surface of a tube or it might be packed into a column. The mobile phase might be a liquid or gas
and also any of thousands of choices.
In this experiment, the stationary phase is a strip paper. The compounds that will move across the paper are food
dyes- specifically the coating on M&M candies. The solvent is salt-water. You should find that food dyes may be a
single molecule to achieve the desired color, or a mixture of dyes. In fact one of your goals is to characterize each
M&M dye to identify the components. For instance, does a green M&M have a single green dye or is it a mixture of
a blue dye and a yellow dye? Under a particular set of conditions, a compound will move a certain percentage of the
path. Thus having a known compound and an unknown compound developed side-by-side can be used to test the
identity of the unknown compound. When the unknown moves the same distance as the known, it is good evidence
that the compounds are identical. The unknown compound can even be just one compound in a complex mixture,
although there are times when a complex might form between two compounds in a mixture that would interfere with
its mobility.
Some might argue that artificial colors have no place in foods since they are unnecessary nutritionally. However the
visual appeal of food is important. Would you choose to eat green peanut butter? Would you buy vegetable oil with
blue streaks in it? Of the nearly 100 food dyes in use around 1900, only 9 are currently approved (certified) by the
Food and Drug Administration. These are sorted into water soluble forms (dyes) and water insoluble forms (lakes).
Along the path to this limited selection of dyes there were several tragic instances. In one case a number of children
suffered serious illness from orange and red dyes in popcorn. Other dyes were found to be toxic or lead to birth
defects, heart problems, or cancer. There are numerous colors that are exempt from certification such as caramel
color and turmeric and all other pigments derived from natural sources. Even today there is controversy concerning
food dyes. Two of the 9 dyes are restricted in their use. For instance Red #32 (now called Citrus Red #2) may be
used only for coloring the skin of oranges. Some have argued that food dyes might be linked to conditions such as
hyperactivity or learning disabilities in children although well-controlled studies in the 1970s disproved this.
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F, D &C Yellow Dye #5 is composed
of tartrazine, one of the nine
HO
S
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approved food colors. This dye is
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known to cause allergic reactions in
N
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some people (in the form of hives in NaO
S
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N
less than 1 person out of 10,000). As
such, a warning label on M&M
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packages informs consumers that
F, D&C Yellow #5 (tartrazine)
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Yellow #5 is one of the colors added,
although it does not specify which colors of M&M's contain this dye. This is also not the only yellow dye listed on
the ingredients (just because you see something yellow does not mean it is yellow #5). Your goal is to use paper
chromatography to test each M&M color to:
(1) determine what dyes compose each color and,
(2) using authentic tartrazine as a reference, determine which colors contain tartrazine, and which contain yellow
dyes that are not tartrazine.
(3) double check your identification by noting the fluorescence color of each dye spot

Compounds that fluoresce emit light of a different wavelength than the wavelength that is shined upon them. For
instance some laundry detergents contain materials that fluoresce such that when UV wavelengths shine on these

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compounds, visible light is emitted. When properly designed this leads to the whiter than white concept. Some
food dyes also fluoresce under 365-nm UV light- sometimes in a similar color to the overall color of the dye, and
sometimes different. For instance Yellow #6 does not fluoresce, while yellow #5 fluoresces a light yellow. Blue #1
fluoresces pink while blue #2 fluoresces blue. Thus you will shine a 365-nm UV light on your chromatogram and
observe which ones fluoresce and in what color.
Procedure
DO NOT EAT THE M&Ms.
Prepare the paper
A square piece of chromatography paper or filter paper is cut to approximately 7 or 8 cm on each side. Along one
side, about 1 cm from the edge, 7 tracks are marked using a pencil. Label each mark with a code for the 6 M&M
colors (red, orange, yellow, green, blue, brown) plus the 7th for the standard tartrazine.
Spot the compounds
For the M&Ms: Dip a toothpick in water and rub it on the shell of the M&M to extract the dye. Take care not to
dissolve so much away that you reach the chocolate. Blot the toothpick lightly on a paper towel and then use a
quick motion to spot the extract on its mark on the chromatography paper. Do this for all of the colors and allow
the spots to dry. Compressed air may be helpful. Repeat this procedure 3 or 4 times by applying another spot on top
of the first until the color is strong.
For the food coloring and tartrazine standard: since this solution will be more concentrated, only spot once with
the toothpick.
Make the paper free-standing
Curve the paper into a half-cylinder and use a piece of tape near the top to help hold it in this shape. Do not roll it
completely up into a full cylinder since you do not want the two vertical edges to touch.
Prepare the developing chamber
Pour enough 0.1% aqueous sodium chloride solution into a beaker such that the level is:
low enough that all your spots on the chromatography paper will be above the surface and
high enough that you will not go dry when the solution is soaked up by the paper.
Develop the chromatogram
Place the paper in the beaker such that it does not touch the sides and the solvent is soaked up evenly (the solvent
front is not crooked). Once the top of the solvent is approximately 2 cm from the top of the paper, remove the paper
and use a pencil to immediately mark the location of the top of solvent and outline the location of each spot. Since
the water will continue to soak up, the solvent front and spots will change location over time. You will use these
initial locations that you mark.
Check the fluorescence colors
A 365-nm UV light will be set up in the darkened lab room across the hallway from your lab. Shine the UV lamp
onto the chromatogram paper and record the results. Do not shine the UV light directly into your eyes or
intentionally on your skin.
Results
Sketch your chromatogram in your notebook. Measure the distance from the starting point to each spot (be
consistent- you probably want to choose the uppermost part of each spot since the spots are likely to tail
significantly) as well as the distance to the solvent front. Use these measurements to calculate the Rf of each spot
according to the formula:
Rf= distance spot moved/distance solvent front moved
The Rf is therefore a ratio (with no units) and will be a number between 0 and 1. Make a table that lists the Rf value
for each spot for each color. Obviously the colors that consist of several dyes will have several spots, each with its

own Rf value. If any spots have the same Rf as the tartrazine standard, then you have a good reason to believe that
spot is tartrazine. Make a neat table that summarizes your results in an easily readable form.
Conclusions
In several sentences summarize your conclusions about the dyes that make up each color and evidence that supports
your conclusions.. Use the Rf values to suggest which colors contain yellow dye #5 (tartrazine). You will turn in
your notebook pages as your report for grading.