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Chromatology of Food Dyes in a Candy Lab Report by Kat Currie, Jon Moscot, and Dori LansbachIntroductionKatIn this weeks

experiment, our lab group sought to find out which FD&C dyes were in certain dyes through chromatography. Chromatography involves taking a mixture and separating it into its different components. Chromatography has been used for such things as finding concentrations and chemical fingerprinting, such as detecting drugs in urine. In our experiments we used paper chromatography, but other scientists also use column chromatography with either gas or liquid as their mobile and stationary phases. Scientists are looking for the best way to separate their mixture so using liquid or gasses in the experiment all depends on the mixture being studied. With column chromatography, the mixture is inserted into the column into the mobile phase. Then depending on the interactions with the stationary phase, the mixtures components are separated and then detected. In our experiment, we placed the different dyes on chromatography paper and then saw the different distances the dyes traveled up the paper when it was placed in a beaker with NaCl. Our goal was to determine which FD&C dyes were in each dye and to be able to compare the Rf values of each. We hypothesized that it would be the M & Ms that would contain multiple FD&C dyes over the food coloring dyes.Experimental Dori The U.S. Food and Drug Administration approves only seven synthetic dyes that can be used in food, drugs, and cosmetics. The FD&C dyes are Blue #1, Blue #2, Green #3, Red #3, Red #40, Yellow # 5, and Yellow #6. These dyes are impure, meaning they are mixtures. In order to separate the components of the mixtures we had to use the method of chromatography. Chromatography is a technique for the resolution of mixtures, relying on the differential affinities of substances for a stationary adsorbing medium through which the mixture passes and a gas or liquid mobile medium which carries the components of the mixture (Green). In order to identify which FD&C dyes were present in each color of the M&Ms and which FD&C dyes were present in each color of food we first had to gather the proper materials to conduct the experiment. The materials included were an 8x7 Whatman #1 chromatography paper, 0.1% w/w aqueous solution of sodium chloride developing solution, toothpicks, ruler, paper towels, beaker, Pasteur pipette, pencil, two M&M candies of each color: brown, red, blue, orange, green, and yellow, the food colorings: blue, orange, red, green, and yellow, 0..01 M stock solutions of the FD&C dyes, and a well plate. The first thing we did was to put all the food coloring dyes, FD&C dyes, and M&Ms (after we watered them down and took some of the dye) into the well plate. In row A of the well plate was FD&C dyes Blue 1, Red 40, Yellow 5, Red 3, and Blue 2. Row B had Yellow 6, Green 3, 2

M&Ms (red and yellow). Row C had the food coloring dyes: red, green, yellow, and blue, and Row D consisted of the rest of the M&Ms: green, blue, brown, and orange. Then we poured less than 1cm of the aqueous solution into the beaker because the origin line on the chromatogram will be at 1cm, and it is vital to the experiment that the solution does not touch the origin line when the chromatogram is first placed in it. We then drew the origin line on the chromatograms and put four FD&C dyes, food coloring dyes, and the M&M dyes on one of the four dash marks we drew on the origin line. The first chromatogram consisted of Blue 1, Red 40, Yellow 5, and Red 3, the second chromatogram had Yellow 6, Green 3, the red M&M, and the yellow M&M, the third one had the red, green, yellow, and blue dye, and the fourth had the green, blue, brown, and orange M&M. We had to make a separate chromatogram for the Blue #2 because we forgot to do that one with the rest of the FD&C dyes. Next we placed the chromatograms individually into the beakers of solution and waited about two minutes for the dyes and M&M dyes to rise, until they stopped. For each chromatogram we took them out of the beaker, drew a line in pencil where the solvent stopped, and put them in the oven to dry. It was then necessary to find the retention factor (Rf), or the distance the dyes traveled up the chromatogram. The retention factor is found by taking the distance traveled by the solute and dividing it by the distance traveled by the solvent. By finding that data we were then able to compare the Rfs of the food dyes and M&Ms with the Rfs of the FD&C dyes to determine which FD&C dyes were present in which food coloring dyes and M&Ms. Data DoriThere were 5 chromatograms with four dots that we drew on the origin line where we placed each FD&C dye, food coloring dye, and M&M. We labeled each of the FD&C dyes, food coloring dyes, and M&M's with a letter (A-Q). The first contains the FD&C dyes, the second has two FD&C dyes then two of the M&M's, the third has four of the food coloring dyes, and the fourth chromatogram has the rest of the M&M's.Chromatograms1.) Blue #1(a), Red #40(b), Yellow #5(c ), Red #3(d)2.) Yellow #6(e), Green #3(f), Red M&M(g), Yellow M&M(h)3.) Red chromatogram, and the distances all the different dyes and M&M colors traveled.1.) Solvent - 6.9cm, Blue #1 - 6.9cm, Red #40 - 2.4cm, Yellow#5 - 4.8cm, Red #3 - 1.2cm2.)Solvent - 6.1cm, Yellow #6 3.3cm, Green #3 - 6.1cm, Red M&M - 1.5cm, Yellow M&M 3.7cm3.) Solvent - 6.5cm, Red - 1.2cm (bottom), 4.1cm (top), Green - 5.2cm (bottom), 6.5cm (top), Yellow - 2.2cm (bottom), 5.5cm (top), Blue 2.3cm (bottom), 6.5cm (top)4.) Solvent - 6.3cm, Green M&M - bottom color (yellow) 3.5cm, top color (blue) 5.3cm, Blue M&M - 5.9cm, Brown

(i), Green (j), Yellow (k), Blue (L)4.) Green M&M, Blue M&M, Brown M&M, Orange M&M 5.) Blue #2 (q)Data of the distance traveled of the solvent up the

M&M - bottom color (pink) 1.8cm, top color (yellow) 2.8 cm, Orange M&M 2.9cm5.)Solvent - 6.3cm, Blue #2 - 1.3cmCalculationsDoriThe one calculation that we used in the experiment was one that determined the retention factor, denoted Rf (subscript f) and it was found by taking the distance traveled by the solute and dividing it by the distance traveled by the solvent.Rf (a)= 6.9cm/6.9cm= 1.0cmRf (b)= .35cmRf (c )= .70cmRf (d)= .17cmRf (e)= .54cmRf (f)= 1.0cmRf (g)= .25cmRf (h)= .61cmRf (i) bottom= .18cm Rf (i) top= .63cmRf (j) bottom= .80cm Rf (j) top= 1.0cmRf (k) bottom= .34cm Rf (k) top= . 85cmRf (l) bottom= .35cm Rf (l) top= 1.0 cmRf (m) bottom= .56cm Rf (m) top= .84cmRf (n)= .94cmRf (o) bottom= .29cm Rf (o) top= .44cm Rf (p)= .46cmRf (q)= .21cmData ResultsDoriWe compared the retention factor values of the FD&C dyes to the food coloring dyes and the M&M's to see which FD&C dyes were used in each food coloring dye and each M&M. Whichever food coloring dyes and M&M's Rf values were closest to the Rf values of the FD&C dyes meant that those FD&C dyes were used in those particular food coloring dyes and M&M's.Red M&M - Blue #2Yellow M&M - Yellow # 6, Yellow #5Green M&M - Yellow #6, Yellow # 5, Blue #1Blue M&M - Blue #1Brown M&M - Red #40, Yellow #6Orange M&M - Yellow #6Food Coloring DyesRed - Red #3, Yellow #5Green - Blue #1, Green #3Yellow - Red #40, Yellow #5Blue Red #40, Blue #1DiscussionJonIn our findings, using paper chromatography we found that the FD & C synthetic food coloring traveled very well through the porous surface and there were no physical reactions in the stationary phase that would have split the colors up. Capillary action drew the dye closer and closer to the top, but there was never any change in how weak or strong the analytes interaction was in the stationary phase. Generally our findings supported the fact that the FD & C dyes were very stable and had a long length in the stationary phase. Now if we go ahead and look at the other dyes in this investigation that we would use for cooking in meals at home or that we may find in everyday food the findings are a little bit more interesting. Taking a look at the data we can see that these dyes overall had a farther length in cm than did either the M & Ms or the FD & C synthetic dyes. The reason that we believe this happened is because these food coloring samples have large traces of the synthetic food dye in them; this is not a new fact. It has been known and astonishingly approved by the FDA for there to be small traces of the FD & C synthetic dyes in food and food coloring. What is amazing is that there have been tests ran on these synthetic dyes to see what they have in them and what they have found is that in multiple cases there are extremely harmful effects on humans in larger quantities. For example Yellow 5 has tartrazine in it, which is derived from coal tar and can cause a severe reaction to those people that suffer from asthmatic symptoms. Red 3 contains erythrosine, another compound that is coal tar based and has been recently thought to be a

carcinogen causing thyroid cancer to people when administered in large doses. Now what does this have to do with the investigation and how have we proven this? The answer lies in the chromatography. When we go ahead and look at our data from the 5 separate sheets that we carefully measured and found the Rf value for, we find that the colors began to split up into two separate lengths in the stationary phase and ultimately completely separated from each other. In the Red Dye, we can find the colors pink and red and by comparing the Rf value of the synthetic dyes to the Rf values of the red, yellow, blue, etc we can establish a relationship and say that because the numbers are so close Red Dye has traces of both Red 3 and Yellow 5 in it. S\A few others are Green Dye had Blue 1 and Green 3, Yellow dye had Red 40 and Yellow 5 and the Blue Dye had Red 40 and Blue 1. The M & Ms were a little bit different because they were different by color. Some colors only had one synthetic dye in it whereas others would have 2. The Green and Brown M & M both had 2 sets of synthetic dye in them based off of the length and color line they left in the paper chromatography investigation. This is an interesting phenomenon, the food that we are eating has these known harmful chemicals in them and they are allowed because they are in small traces, something I dont necessarily agree with. I believe that there is a minor flaw with the chromatography in this investigation. I think that one way to make this experiment more accurate would be to use a couple more ways to judge exactly where each part of the analyte begins and ends so if there were mixed colors it wouldnt be so much of a guessing game, but more of a precise equation. If we were to find the polarity, hydrophobicity and hydrophobility I feel like this experiment would be a lot more accurate than it already was.ConclusionKatAt the conclusion of our experiment, we discovered that our hypothesis was indeed supported by the results. The chromatograms showed that the FD&C dyes were present in the M & Ms and the food coloring. This makes sense, as the only dyes approved for use in food are the FD&C dyes we tested. It was hard to distinguish some of the dyes based on Rf value because Blue 1 and Green 3 had the same value. This shows that there must have been some experimental error. Also, the M & Ms showed they contained a lot more mixture in the different FD&C dyes in their composition. The investigation was very interesting as it allowed our group to try the new experimental technique of chromatography. ReferencesGeneral Chemistry Online! Senese, Fred. Revised: 25 July 2005. http://antoine.frostburg.edu/chem/senese/101/matter/chromatography. shtmlPost-Lab Questions1. There is a big difference in the ingredients and there effect between the movements of the food dyes and the M & Ms on the chromatography paper. The M & Ms did not travel on average near as far up the chromatography paper as the food dyes did. The ingredients in the food dye were limited including the dye,

water and propylene glycol. The M & Ms included sucrose, corn syrup, cornstarch and gum acacia, all of which add onto the weight of the analyte and slow down the process of chromatography in the stationary phase. The less ingredients that there are, the better the solvent will travel with the solution and our data supports this claim. Our Rf values for the Dyes at the top of the line read .63 cm, 1.0 cm, . 85 cm, and 1.0 cm. For the 6 M&Ms there were only 2 M&Ms that had an Rf value higher than .63 cm and 1 that was higher than .85 cm. Without knowing the data that says there are more ingredients in M&Ms, it would have been safe to make that guess based on the Rf values and also just by looking at the chromatography paper and seeing the faintness and length of their lines.2. The color food dyes that should not be eaten or used in any capacity by someone that is allergic to tatrazine, an extract from coal tar are the Red dye and the Yellow dye. Both of these dyes have Yellow 5 in them, which is tartrazine along with Red 40 and Red 3; Red 3 is a carcinogen.3. The Yellow M&Ms and the Green M&Ms should not be consumed by the persons allergic to tartrazine because they both contain the FD&C dye Yellow #5.4. The color blends of the green, orange, red, and yellow food dyes and M&Ms are for the most part different. There are some similar colors for example the red and yellow food coloring dyes both contain Yellow #5 and so do the Yellow and Green M&Ms, but other than that the makeup of the food dyes and the M&Ms are different.5. Of the seven FD&C food dyes, six are present in the samples tested in this experiment. They are Blue 1, Red 40, Yellow 5, Red 3, Yellow 6 and Green 3. Blue 2 is the only one not present in the samples.6. We were able to separate the different dyes using this technique because of the NaCl that we put in the bottom of the beaker. When we placed the chromatology paper inside the beaker touching the bottom, the NaCl traveled up the paper through capillary action. This capillary action carries along the different parts of each of the dyes. On the way up, the specific dyes stopped at certain places along the paper. This left distinct streaks of color on each paper.