Crime scene investigation labIdentification of Inks in Felt Tip Pens Using Paper Chromatography

Background A crime has been committed. A wealthy celebritys prize winning Poodle Fluffy has been kidnapped. At the scene of the crime was a handwritten ransom note. In it the kidnapper demands $1 million dollars or The pooch gets it!!! The celebrity believes that a disgruntled former employee is behind the kidnapping. Due to your well known expertise in such matters, she has hired your services to find out who the culprit is. You sneak into the three suspects offices late at night. You then borrow pens/markers from these three suspects desks. You will compare the ink in these markers to that of the ransom-notes. You will use paper chromatography to determine who wrote the note. Theory During its development over the last 40 years, chromatography has revolutionized modern analytical chemistry. Chromatography is a group of techniques used to separate colored mixtures into their component parts. Many types of chromatography are now routinely used in la oratories around the world to separate and identify components in mixtures. !he analyses of lood and urine samples for drugs and the analyses of drin"ing and ground water for hazardous chemicals are two common procedures involving chromatographic separations. #ll chromatography techniques involve a stationary phase and a mobile phase. !he stationary phase can e either liquid or solid. !he mixture to e separated is usually placed on the stationary phase. !he mo ile phase can e either liquid or gas. !he mo ile phase moves along the stationary phase, carrying some or all of the mixture with it, resulting in the separation of the mixture components. $n liquid chromatography, separation is ased on the preferential attraction of each component in the mixture to either the mo ile or stationary phase. !he attraction is due to

particular intermolecular interactions. %aper chromatography is the simplest form of chromatography. #lthough paper chromatography is not used for drug or hazardous chemical analyses, it is extremely useful for separation and identification of food dyes. $n paper chromatography, a sample of the mixture to e separated is placed on a piece of chromatography paper, which acts as the stationary phase. &ne edge of the paper is placed in a solvent, such as water, alcohol, or a mixture of oth, which acts as the mo ile phase. Many different solvent systems are possi le, depending on the components to e separated. !he chromatography paper acts li"e a wic", drawing the solvent up the paper y capillary action. !he wic"ing occurs ecause the solvent is attracted to water molecules that are permanently ound to cellulose fi ers of the paper. !he water molecules ound to the paper and the paper itself forms the stationary phase. # sample is applied, or spotted, on an area of the paper near the ottom edge, "nown as the origin line. !he ottom edge of the paper is then placed in a solvent, and solvent is drawn up the paper. 'hen the leading edge of the mo ile phase, the solvent front, reaches the sample, the sample components are preferentially attracted to either the stationary or mo ile phase. !his attraction depends on the relative polarities of the sample components and the two phases. (ecall that li"e solvents dissolve li"e solutes. )owever, the attraction is seldom an all*or*nothing situation. Most compounds, whether they are ionic or molecular, are somewhat attracted to oth phases. +quili rium is esta lished for each component etween the two phases, as shown y +quation ,. component*mo ile phase component*stationary phase -+q. , .

#s the solvent front moves up the paper, fresh solvent passes the spotted sample, and new equili ria are continually esta lished. #t the same time, any components that have dissolved in the mo ile phase encounter fresh stationary phase, and new equili ria are esta lished. !hus, the components of a mixture move up the paper at different rates and separate, producing a pattern called a chromatogram. !he overall effect of these equili ria is that the movement of the components depends directly on their relative attractions for the mo ile and stationary phases. 'e characterize this movement in terms of a retention factor -(f. defined y +quation /.

distance traveled y component, cm (f 0 ************************************************ distance traveled y solvent front, cm

-+q. /.

$f a component moves with the solvent front, its (f value can e as high as ,.0. $f the component does not move at all, its (f value can e as low as 0.0. !he (f value for a component is reproduci le for a particular component*solvent system, if the experimental conditions are closely controlled. &ne important varia le is the composition of the solvent. $f one of the solvent components is volatile, the percent composition of the solvent may change during the analysis, due to evaporation. !his will affect the results. Covering the chromatogram developing container so that the air in the container remains saturated with solvent vapor can prevent evaporation. # sample containing two or more components can e separated, or resolved, if we choose a solvent system for which the sample components have distinctly different (f values. $n order to choose the most appropriate solvent, we first determine the (f values of the individual sample components in a variety of solvent systems. !hen we choose the est solvent system for separating all of the sample components. 1igure , illustrates the preparation of a chromatogram. 2pots of the sample to e resolved are placed on the origin line of the chromatography paper and the ottom edge of the paper is placed in the solvent. 2olvent moves up the paper separating the components. !he distance a component has moved is determined y measuring the distance from the origin line to the center of the component spot. $n 1igure ,-c., the left*hand spot is the sample eing resolved and the right*hand spot is Compound #, a component that we thin" is present in the mixture. !he distance the solvent front moved is 4.0 cm, and the distance the middle component of the mixture moved is /.0 cm. !he middle component3s (f is /.0 cm / 4.0 cm 0 0.40. !he distance Compound # moved is /.0 cm, so its (f is also 0.40. &n the asis of identical (f values, we can conclude that Compound A is pro a ly one of the mixture3s components. 1requently, other information from chromatograms supports findings ased on (f calculations. 1or instance, when resolving food dyes, we can compare the colors of the spots. 1or example, when resolving green food coloring, we o serve two spots, one yellow and one lue. !hese colors, in con5unction with the Rf values of the components, help identify the dyes in the green food coloring.

1igure ,.

1elt tip pens have a 6rod7 inside them. $t7s made of an a sor ent material and is impregnated with a mixture of different colored dyes. !he color you get from a pen depends on the mixture of in"s used. $n" dyes can e separated using paper chromatography. !here are two types of pens 8 permanent and non*permanent. !he difference is how easily they can e washed from what they have een used to mar" -whether deli erately or accidentally..

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2cope

!he procedure descri ed elow may e used to identify the in" dyes used in non* permanent and permanent felt tip pens. /. %rinciple

2u stances are separated in chromatography ecause of the difference in their attraction for the mo ile phase -the solvent. and the stationary phase. !he greater the attraction for the stationary phase relative to the mo ile phase, the slower a su stance moves through it. 9. (eagents: 'ater, $sopropanol -)$;)<= 1<#MM#><+? $(($!#@!..

4. +quipment: <arge test tu es, 2cissors, (uler, !est*tu e rac"s, Chromatography paper strips. 4. )ealth and safety: 'ear protective clothing and eye protection.

A.

Procedure

* %ieces of chromatography paper will e provided, use one for each pen to e tested. !he 6ransom note sample7 -or +vidence. will e prepared for you. !he 6evidence7 will have a code written on it, e sure to record this on your data sheet. * %lace test tu es -four in total, one for each piece of chromatography paper. in a test* tu e rac". Mar" the test ransom note test tu e with 6+7 for evidence and other test tu es will e mar"ed ,, /, and 9 using a mar"ing pencil or 2harpie. * Draw a pencil line a out / cm from one end of each piece of paper. #t the center of the line use a felt tip to ma"e a dot. !a"e care that it doesn7t spread out too much. $deally it should e a dar" dot no more than / mm across. * 1old the chromatography paper at the opposite end of the dot so that it can rest over the edge of the test tu e and support the strip. * %lace a out , cm depth of water in each tu e.

* Carefully insert the chromatography paper into the test*tu e, dotted end down. !he dot must e a ove the water, and the sides of the chromatography paper should not touch the sides of the test*tu e.

* <eave the test*tu e until good separation is o tained or until the solvent front reaches ,*/ cm from the top of the chromatography paper. * * (emove the chromatogram and hang it to dry. Mar" the solvent front with a pencil and put a small cross at the centre of each spot.

* (epeat the experiment using a ,:, -volume: volume. mixture of isopropanol and water as the solvent

7.

Analysis and calculations

At the center of each spot that you observe, place a dot with a pencil. There may be more than one spot for a given sample.

Calculate the R f value and give the color for each spot that appears on the chromatograms

Identification of inks Data Sheet / Post-lab Questions NAME:___________________________ PARTNER:________________________ DATE: ______________ SECTION: ___________

Compile a listing of all spots that you observed for each sample, their R f values, and the color of the spot (such as red, pink, orange, yellow, purple, green, or blue ) Suspect No.1 (you may not have three spots!) Rf water color Rf water +isopropyl alcohol

color

Suspect No.2 Rf water

color

Rf water +isopropyl alcohol

color

Suspect No.3 Rf water

color

Rf water +isopropyl alcohol

color

Crime Scene Evidence Rf

water

Code #: ____________ Rf water +isopropyl alcohol color

color

Verdict Based on the evidences you collected, which suspects are innocent? Which suspect(s) must be investigated further? Explain.

Likely innocent:

Remains suspicious

Identification of inks Pre!lab "ssignment NAME:___________________________ DATE: ______________ SECTION: ___________

1) Briefly explain the problem that will be created if you mark the original baseline with a pen instead of a pencil.

2) The chromatogram shown below is done for four compounds A ~ D, and two mixtures containing compounds A - D. Calculate their R f values. Compound A The R f value = Compound B The R f value = Compound C The R f value =

Compound D The R f value =

Mixture #1 The R f value = The R f value = The R f value = Mixture #2 The R f value = The R f value =

3) Based on your R f values and the appearances, which mixture is contaminated with an additional substance other than compounds A - D?

solvent front

mix#1

mix#2

baseline