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JJC CSI

PURPOSE To learn about the technique of paper chromatography. To develop a paper chromatography technique to separate components in an ink sample. Identify an ink sample based on its chromatogram. Write a report that could be used as evidence in a criminal trial.

INTRODUCTION Paper chromatography was developed in the early 1940s in England. The technique is based on the fact that paper contains a thin film of water around the cellulose fibers of the paper, called a stationary phase. A mixture of the components to be separated is placed in a small spot at one end of the strip of paper and an organic solvent (the mobile phase) is passed over the spot and across the paper. Since each compound present has a different size, shape, and polarity, each compound will dissolve in the water and organic solvent to a different extent. The net result is that if two compounds are started at the same place and solvent passed over them, one compound will move along the paper faster than the other. After a period of time the flow of the mobile phase is stopped. The paper is dried and then sprayed with a reagent that will produce colored spots, if the compounds are not colored. The material used in our experiment is already colored, so the latter step is not required. Chromatography works because of differences in polarity between the stationary phase, the solvent, and the components of the mixture being studied. When using a solvent with low polarity, the majority of the components of the mixture, if not all of them, will be held more strongly to the stationary phase than the solvent molecules, so the solvent will not displace them and carry them along with it. If a more polar solvent is chosen, it will displace the majority of the molecules in the sample, but depending on the polarity of each of the components, some may travel farther from the solvent than others, resulting in separation of each of the components. The distance traveled with the solvent are characteristic of different molecules and make identification possible. A solvent much more polar than the components of the mixture will displace all of the molecules and carry them all very easily on the solvent front, yielding no separation that would be useful in identification. Finding a solvent suitable for separation of a mixture or identification of a sample usually involves trying several solvents/solutions of different polarity. In your experiment you will be using deionized water (most polar), a 20% ethanol solution, a 50% ethanol solution, and a 90% ethanol solution (least polar). Criminal investigators use chemistry routinely to solve crimes. Sometimes crimes involve the use of ink pens. Forgery cases, kidnapping cases (involving a ransom note), and other crimes involving writing are often investigated using chromatography. Someone has kidnapped a beloved pet fluffy, leaving a ransom note at the scene of the crime. The CSI team has collected black pens from the suspects. In this investigation,

your team will be given a sample of the ink found at a scene of the crime. Your job will be to determine which pen was used to write the note. MATERIALS Black ink pens Chromatography paper Scissors 4- 600 mL beakers Rulers PROCEDURE Black ink is usually a mixture of several different colors that can be separated that can be separated by chromatographic conditions. Designing chromatography experiments usually involves selection of the proper solvent for the system of interest. In this investigation, your group of crime specialists has been provided several solutions of ethanol with water. Your first task should be to determine which solvent mixture is appropriate for separation of ink components in the known pens. In addition to the ethanol solutions, you should try using deionized water as the solvent. In addition to determining the proper solvent, you should develop a way of quantifying your data for your report; that is, you might consider ways of expressing your results in numerical terms that allow for easy comparison of results from one experiment to another. Parafilm squares Ink sample from crime scene Deionized water 20%, 50%, and 90% ethanol solutions

This experiment is best done in groups of four individuals. Expect further detailed directions from your laboratory instructor about how you will go about the labs activities. 1. Obtain four pieces of 16 cm x 11 cm piece of chromatography paper. Use a pencil to draw a line along the 16 cm side of the paper that is 1 cm from the bottom of the paper. Fold the paper as you would a fan to generate 5 panels as shown below. 2. Use each of the confiscated pens to place a small dot (approximately 3 mm wide) of ink on each panel. Use a pencil to label the identity of each ink sample as shown below. It is OK to use abbreviations as long as you provide a key to what they are in your report. Allow the ink samples to dry for about one minute.
fold in paper

pencil line Flair Sharpie Vis-aid Vis-a-vis Calligraphic

3. Collapse your fan of paper slightly so it will stand up in a beaker on its own and so the sides of the paper will not touch the walls of the beaker. 4. Pour 20 mL of deionized water in a 600 mL beaker. Do not use a bigger or smaller size beaker. 5. Insert the folded chromatography paper into the liquid. The pencil line end of the paper should be closest to the liquid. Be sure the ink dots are above the liquid level. 6. Remove the protective paper from the square of parafilm. Keeping the beaker on the bench top, carefully stretch the parafilm over the top of the beaker to seal it. Do this very carefully so that the piece of chromatography paper is not disturbed. 7. When the liquid has nearly, but not completely reached the top, remove the paper, use a pencil to mark how high the solvent traveled. Allow the chromatography paper to air dry before making any measurements. 8. Repeat the same procedure for the other solutions. Each member of the group should be responsible for using one of the solvents/solutions. 9. Your group will be given one or two samples of ink taken from a crime scene. Because the sample is taken from a limited supply that will be used in court, do not use the unknown until your group is certain of the solvent system you will use. You are looking for the solvent that will allow you to easily identify each sample. 10. The set up for the unknown is a little different than what you used for the large pieces of chromatography paper.

11. Place approximately 1 cm of the solvent of choice in a 250 mL erlenmeyer flask from you equipment drawer/tote tray. 12. Place the paper with the ink spot end closer to the liquid. Allow the paper to touch the liquid, but not the bottom of the flask. Carefully insert a cork or stopper in the flask to hold the paper in place. When the liquid has nearly, but not completely reached the top, remove the paper and allow it to air dry. Cleanup: All the solutions used may be disposed of down the drain. Thoroughly rinse and dry all the 600 mL beakers before returning them. Report The report must be typed and grammatically correct. It may be returned to you for corrections if it is not acceptable. Your report should include a discussion of factors that may affect the rate at which the solvent and the ink components move up the chromatography paper. Remember that your report may be used in a court of law. If directed by your instructor, include your chromatograms with your report.

QUESTIONS (Please put answers in your lab report) Q1: What pen was used to write the ransom note Q2: Why do you suppose some solvent systems work better than others do in the separation of the components of ink? Q3: Why should the solvent depth of the chromatography solvent remain below the level of the ink dots? Q4: Some components of ink are minimally attracted to the stationary phase and very soluble in solvent. Where are these components located on the chromatography paper during chromatography?