Chromatography of Food Dyes Introduction Chromatography deals with a series of techniques that are used to separate components of mixtures.

In each case a mixture of solutes dissolved in a moving solvent, called a mobile phase, passes over a selectively absorbing stationary phase. The mobile phase is a liquid or a gas, and the stationary phase is a solid. Separation takes place because the components of the mixture have different attractions for the two phases and move through the system at different rates. There are a number of chromatographic procedures that can be used. In thin layer chromatography !T"C#, the mobile phase moves across a stationary phase that contains a thin layer of particles spread on a glass plate. Column chromatography makes use of a stationary phase of small particles packed into a tube. The mobile phase, which can be either a liquid or gas, passes through the solid phase. In this experiment, you will use paper chromatography. $aper forms the stationary phase over which a mobile solvent moves. %hen the components that are to be separated are spotted on the paper chromatogram, the bottom of the paper is submerged in the developing solvent. The solvent moves up the paper by capillary action, and the separation process takes place. &nce the chromatogram has been developed the retention factor !'f# value is calculated. The 'f value is the distance traveled by the solute compound divided by the distance traveled by the solvent front. There are only seven food drug and cosmetic !() and C# dyes that have been approved by the (ood and )rug *dministration !()*# for use in foods. They are shown in the table below along with their dye name and color+ Dye Number ,lue -o. . ,lue -o. / 'ed -o. 0 'ed -o. 23 4ellow -o. 5 4ellow -o. 7 8reen -o. 0 Dye Name ,rilliant ,lue (C( Indigotine 1rythrosine *llura 'ed *C Tartra6ine Sunset 4ellow (ast 8reen (C( Color ,right ,lue 'oyal ,lue $ink &range red "emon 4ellow &range Turquoise

The purpose of this experiment is to measure the 'f values of the ()* approved food dyes, and then identify by measuring the 'f values and observing their colors, dyes found in foods that are not known. The 'f values and colors of ,lue -o. / and green no. 0 will not be determined because they are seldom used in foods. Pre-Lab Problem: *fter the development of a chromatogram, it is found that a dye compound moves 7.. cm and the solvent front moves .3./ cm. %hat is the 'f value of the dye9

Procedure: .. /. &btain four pieces of chromatography paper !or %hatman -o. . filter paper# that have been cut into ./ x .2 cm rectangles. :sing a pencil and ruler draw a line / cm above the bottom of the long edge of each of the filter papers. )on;t use ink to draw the line because ink contains dyes that will be moved by the solvent. $encil in five dots on the lines about ../ cm apart that are large enough to be seen above and below each line. *pply spots of blue no. ., red no. 0, red no. 23, yellow no. 5 and yellow, no. 7 dyes to the dots on two of the filter papers with toothpicks, and let the dyes dry for at least 5 minutes. This is done to allow the solvents that the component dye compounds are dissolved in to evaporate. Spot a sample of each unknown dye mixture on each of the two remaining filter papers. Spot them on the penciled dots. "et the samples dry for at least 5 minutes. ,end each of the filter papers into a cylinder along the long edges and staple the edges together at the top and at the bottom. $our /3 m" of 3..3= -aCl in water !the developing solvent# into each of four 733 m" beakers. Set the filter paper cylinders down into the beakers containing the developing solution. The spotted dye samples must be above the solvent in the beakers. *llow the solvent to move up the filter papers and develop the chromatograms. "et the solvent reach within / cm of the top of the filter papers and remove the chromatograms from the beakers. @ark the positions of the solvent fronts with a pencil line before all of the solvent evaporates. These will be the wet leading edges of the solvent. &nce the paper has dried, find the densest part of the band for each colored spot and draw a circle around it with your pencil. Then put a dot in the center of the circle. @easure the distance !in tenths of a centimeter# from the pencil line at the point where each colored dye or unknown sample was spotted to the densest point of the band of color.

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-ext measure the distance from the pencil line at the point where each colored dye mixture was spotted to the line for the solvent front. Calculate the 'f values for each colored dye compound !each spot# for both the known dyes and unknown dye mixtures and record them on your data sheet. &btain an average 'f value for each dye. Identify the dyes found in your unknown dye mixtures from their colors and 'f values.

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Data Sheet -ame AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA I. *. Bnown (ood )yes ,lue -o. . Trial . )istance traveled by dye compound )istance traveled by solvent front 'f value *verage 'f value Color ,. 'ed -o. 0 Trial . )istance traveled by dye compound )istance traveled by solvent front 'f value *verage 'f value Color C. 'ed -o. 23 )istance traveled by dye compound )istance traveled by solvent front 'f value *verage 'f value Color ). 4ellow -o. 5 Trial . )istance traveled by dye compound )istance traveled by solvent front 'f value *verage 'f value Color Trial / Trial / Trial /

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4ellow -o. 7 Trial . )istance traveled by dye compound )istance traveled by solvent front 'f value *verage 'f value Color Trial /

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:nknown )ye @ixtures *. :nknown C. (irst )ye Trial . )istance traveled by dye compound )istance traveled by solvent front 'f value *verage 'f value Color of )ye Identity of )ye Second )ye Trial . )istance traveled by dye compound )istance traveled by solvent front 'f value *verage 'f value Color of )ye Identity of )ye Third )ye Trial . )istance traveled by dye compound )istance traveled by solvent front 'f value *verage 'f value Color of )ye Identity of )ye Trial / Trial / Trial /

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:nknown C/ (irst )ye Trial . )istance traveled by dye compound )istance traveled by solvent front 'f value *verage 'f value Color of )ye Identity of )ye Second )ye Trial . )istance traveled by dye compound )istance traveled by solvent front 'f value *verage 'f value Color of )ye Identity of )ye Third )ye Trial . )istance traveled by dye compound )istance traveled by solvent front 'f value *verage 'f value Color of )ye Identity of )ye Trial / Trial / Trial /

Post-Lab Questions: .. * green lime popsicle does not actually contain any green dye. 1xplain how the green color could be obtained.

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Dow would you formulate a purple solution from the dyes you used in this experiment9