Aljosa Timarac

Chromatography of M&M
OBJECTIVES - To become familiar with techniques of separating mixtures - To learn about chromatography, specifically paper chromatography

DISCUSSION The word chromatography means color-writing and the name was chosen at the beginning of this century, when the method was first used, to separate colored components from plant leaves. The name chromatography may be misleading, because separation techniques do not depend on color. !any variations of this technique exist today. "hromatography is used as a very reliable and fast technique to separate a variety of natural and synthetic mixtures into their components. #everal types of chromatography exist today, among which are$ paper chromatography, thin-layer chromatography or T%", liquid-liquid chromatography, gas chromatography, or high performance liquid chromatography or &'%". Chromatography is a separation technique used to separate the components of a mixture. !any separation methods rely on physical differences between the components of a mixture. (ndoubtedly, you are already familiar with several means chemists use to effect separations based on physical differences. These techniques include$ Filtration, where separation may be effected because substances are present in different states )solid vs. liquid*+ Centrifugation, where separation is effected by differences in density+ and Distillation, where separation is effected by ta,ing advantage of differences in boiling temperatures of the various components. -n this laboratory exercise, we will effect a separation of a mixture of food dyes using paper chromatography. -n a typical experiment, a mixture of different chemicals is partitioned between a stationary phase and a moving phase )eluent or solvent*. The moving phase may be a liquid or a gas+ the stationary phase is typically a solid. As the mobile phase flows over the stationary phase, the components in the mixture are carried along. The ability to separate a mixture into its components basically depends on two fundamental concepts$ solubility and a sorptio!. The better soluble a component is in the moving phase the faster it will be transported along the stationary phase. Adsorption refers to the ability of a substance to stic, )or be adsorbed* to the surface. The better the component is adsorbed to the stationary phase, the slower it will be transported along the stationary phase. As the mixture moves over the stationary phase, the components in the mixture move further and further apart into discrete .ones.

"ap#r chromatography uses ordinary filter paper as the stationary phase, which consists primarily of cellulose. Thi!$lay#r chromatography )abbreviated T%"* uses a thin glass plate coated with either aluminum oxide )alumina* or silica gel as the solid phase. The moving phase is a liquid and is chosen accordingly with respect to the properties of the components in the mixture. -n a typical experiment, a single drop of the mixture is applied at one end of a strip of filter paper. The filter paper is then placed in a shallow layer of solvent or solvent mixture in a jar or bea,er. #ince the filter paper is permeable to liquids, the solvent begins to rise by capillary action. As the solvent rises to the level at which the spot of mixture was applied, the components of the spot that are completely soluble will be moving along with the solvent upward )refer to /igure 0*. The rate at which a component will move along depends on the relative tendency to be dissolved in the solvent and adsorbed on the paper. 1y this means, the original spot of the %igur# & mixture is spread out. The net effect is that the components #et up for thin layer separate from each other and spread out into a series of bands chromatography or new spots. -t is possible to describe the position of spots )so the substances that have separated* in terms of their retention factor, the 2f value )/igure 3*. The retention factor is defined as$ 2f 4 distance traveled by spot distance traveled by solvent

1ecause the retention factor for a particular mixture may vary depending on the conditions, a sample of ,nown composition is typically analy.ed at the same time on the same sheet of paper or slide. The 2f value is a characteristic property of a given compound in a given solvent in a chromatography experiment. )#ee page 56 of your text for more information on chromatography* %igur# '$ 7xample for calculating the 2f value

-n today s experiment, you will analy.e )not eat+ sorry* the shell coating of !8! candies. #ome !8! s use yellow 9: )tartra.ine* as color. This dye causes allergic

reactions in some people and must therefore be named as an ingredient in products that contain it. The !8! pac,age states that yellow 9: is one of the colors added. The big question is then$ is the dye present in all candies or only in some of the candies of certain colors; The answer to this question is important to the person who is allergic to yellow 9: and wants to ,now which candies are safe to eat. -n the second part of today s experiment, you will analy.e felt-tip pen in,s. /elt-tip pens come in many different colors but most colors are often mixtures of primary color in,s. <e will use water-soluble pens and further study the importance of the proper solvent or solvent mixture in paper chromatography for optimal separation. M(TE)I(*S (ND E+UI"MENT !8! candies 90 chromatography paper or filter paper 3:= m% bea,er =.0 > ?a"l

")E*(BO)(TO), (SSI-NMENT. Chromatography +u#stio!s 0. 7xplain in your own words why samples can be separated into their components by chromatography. 3. <hy must the spot applied to a chromatography plate be above the level of the developing solvent; @. -n a chromatographic experiment, why do some substances move up the stationary phase faster than others; A. -f the solvent front moves B.= cm and a component in a sample being analy.ed moves @.3 cm from the baseline, what is the 2f value; "art I )!8! candies* 0. A 0= x 0=- cm strip is cut from <hatman 90 chromatography or filter paper. Along one side, 0 cm from the edge, draw a straight pencil line, and ma,e seven mar,s at about 0 cm intervals. #ix of these are identified by code for the different colored candies, and the seventh for the standard. The standard will be the compound tartra.ine. 3. To separate the dyes used to coat !8!s, you must first prepare a solution of the coating dyes. To do this, place four !8! s of the same color into a small bea,er. Add @ m% of a :=C:= mixture of water and ethanol and swirl the solvent until the candy coating has dissolved. )#mo/# th# M&Ms from th# sol/#!t b#for# th# chocolat# c#!t#r is #0pos# 1 The dye solutions are not very concentrated, so you will need to spot these dye mixtures several times to obtain a sufficiently dar, spot. Dry each spot between

applications of the mixture to maintain as small and concentrated a spot as possible. Try to ,eep your spots less than Amm in diameter. @. 2epeat the procedure for all color-coated candies. Eently dry the paper by moving it in the air. A. (se a 3:= m% bea,er. Add =.0> sodium chloride, )solvent or eluting solution* so that the bottom of the bea,er is covered. 'lace the filter paper in the eluting solution, with the sample spots down near the liquid surface, but not in the liquid )/igure @*. Tightly seal the bea,er with clear film and let it stand undisturbed. The solvent will gradually rise by capillary action, carrying the components in the spots along. :. The paper is ta,en from bea,er when the solvent front is about 3 cm from the top. 1ecause the paper remains wet for several more minutes and the solvent continues to rise, it is /#ry importa!t to ra2 th# sol/#!t li!# a! circl# th# color# spots imm# iat#ly. 6. !easure the distance from the straight line on which you applied the spots to the solvent front. Then measure the distance from the pencil line to the center of each colored spot and calculate the 2f value from the two number you obtained. 2epeat the procedure for each colored candy spot that you applied. 5. "ompare the 2f values that you calculated to the ones given in the Table to determine which colors are present in each candy that you investigated. 7ach organic compound, and many inorganic compounds, has a characteristic 2f value in a certain solvent. The colors of the shells of ! 8 ! candies contain food coloring that are organic dyes and therefore display characteristic 2f values. #ample Fellow !8! Grange !8! 2ed !8! Ereen !8! 1rown !8! "olors after development yellow orange red yellow, blue blue, red 2f values =.:= =.@: =.03 =.AB, =.5B =.55, =.0@

)#cor i! your !ot#boo3. 2ecord your solvent, distance solvent moved )in cm*, distance color moved )in cm*, and calculate the 2f value for each color. "ompare the 2f values you have obtained with the literature values in Table -.

"art II )/elt tip pens* 0. A 0= x 0=- cm strip is cut from <hatman 90 chromatography or filter paper. Along one side, 0 cm from the edge, draw a straight pencil line, and ma,e four mar,s at 0 cm intervals. Gbtain several water-soluble pens from your -nstructor. Apply a single small spot of different in, to each of the mar,s on the filter paper. Allow the spots to dry completely. Eently dry the paper by moving it in the air. (se a 3:=-m% bea,er. Add the butanol $ water $ ethanol $ acetic acid mixture )03=$A=$3=$0*, which is the solvent or eluting solution. !a,e sure that the bottom of the bea,er is covered. 'lace the filter paper in the eluting solution, with the sample spots down near the liquid surface, but not in the liquid )/igure 0*. Tightly seal the bea,er with clear film and let it stand undisturbed. The solvent will gradually rise by capillary action, carrying the components in the spots along. The paper is ta,en from the bea,er when the solvent front is about 3 cm from the top. 1ecause the paper remains wet for several more minutes and the solvent continues to rise, it is very important to draw the solvent line and circle the colored spots immediately. !easure the distance from the straight line on which you applied the spots to the solvent front. Then measure the distance from the pencil line to the center of each colored spot and calculate the 2f value from the two numbers you obtained. 2epeat the procedure with several permanent in, mar,ers using the same solvent.

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