CHEM 2302 Study Guide - Final Guide: Carnosine, Imidazole, Peptide Synthesis
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Gather 2 pieces of square filter paper, the scissors, a pencil, and a ruler. DO NOT use a pen
or any form of ink. Create a strip of filter paper that measures 14 cm by 7 cm. See Figure 3.
Use the ruler to measure 14 cm across the edge of the paper. Use the pencil to draw a small
âxâ at 14 cm. On an adjacent edge of the paper, use the ruler to measure 7 cm up the edge
of the paper. Use the pencil to draw a small âxâ at 7 cm. Using the ruler as a straightedge,
lightly trace a rectangle that is approximately 14 cm by 7 cm. Use the scissors to cut out the
rectangle. Create two additional strips of paper.
Note:There should be enough filter paper to create a fourth 14 x 7 cm strip of paper if needed.
2.Gather the 25 mL graduated cylinder, 100 mL beaker, distilled water, and salt. Use the
graduated cylinder to add a total of 50 mL of distilled water to the beaker. Add a pinch of salt
to the beaker. Use the stirring rod to stir the solution until the salt has completely dissolved,
resulting in a salt solution of approximately 0.1% NaCl.
3.Gather 2 sheets of the filter paper â 14 x 7 cm from step 1.
a.Create a base line by drawing a thin horizontal line with a pencil (NOT a pen!)
across the paper, 10 mm (1 cm) from the bottom. The line should barely be visible.
b.With a pencil, draw 9 small cross lines along the horizontal line, 1.5 cm apart,
Experiment
Chromatography of Food Dyes
4.Repeat step 3 for the second piece of filter paper.
5.On Sheet 1, from left to right, use a pencil to lightly label each section between the cross lines
with the abbreviation for the six FD& C food colors from your experiment bag that will be
tested: B1, B2, R3, R40, Y5, and Y6.
6.Also on Sheet 1, after Y6, place the labels KG and KS to represent the Kool-
Grape and Kool-Aid
®Strawberry drink mixes to be tested.
7.On Sheet 2 from left to right, use a pencil to lightly label each section between the cross lines
with the abbreviation for the grocery store food coloring that will be tested: SR, SY, SG, and SB.
8.Also on Sheet 2, after SB, place the label âUKâ for unknown and then âMâ plus a letter for the
color of 4 different candies from your bag of M&Ms: such as; M-R (for M&M® red), M-Y, M-B,M-G.
9.Set Sheet 2 aside and perform the following for Sheet 1. After you have completed the
experiment for Sheet 1, repeat for Sheet 2.
10.Put on your safety gloves and goggles.
11.For each dye on the sheet to be tested:
a.Use the scissors to cut open the pipets and place them in the 24-well plate, bulb
side down, using the well plate as a pipet holder.
b.Place a few drops of the dye in a well of the 24-well plate, as shown in Figure 4.
Dye drops and pipets in well
12.Once the paper is spotted with all the dyes, allow the spots to dry for a few minutes.
13.Form the spotted filter paper into a cylinder (colored spots on the inside) with the edges
touching, but NOT overlapped, and staple at the top and bottom as shown in Figure 7. (You
may want to use small pieces of tape on the outside to lightly hold the cylinder together
while you securely staple it. If so, remove the tape after stapling.) Set the cylinder aside for a
moment.
14.The salt water (~0.1% NaCl) you previously prepared will be your eluting solvent and a Petri
dish will be your elution chamber. Pour salt water into the clean and empty Petri dish to a
depth of about 1/4 cm. This is just enough to cover the bottom of the elution chamber.
15.Set your paper cylinder next to (not in) the Petri dish, with the spots at the bottom, and look
to make sure the solvent level is below the base line. If it is not, pour out a little solvent until
it is.The base line (and spots at the bottom of the chamber) must be above the solvent level
for this experiment to work.
16.Carefully place the paper cylinder into the eluting chamber, making sure not to touch the Petri
dish sides. See Figure 8. The solvent-front will travel up the paper rapidly at first and then will
slow down. Let the solvent-front rise, monitoring every few minutes. Immediately remove the
cylinder if any dye or the solvent front moves up higher than 2 cm from the top of the paper.
If all the solvent is soaked up before the front has time to move toward the top of the paper
carefully add a little more solvent to the Petri dish.
17.When the solvent-front has traveled up the filter paper, 2 cm from the top, remove the filter
paper cylinder from the chamber and immediately mark the top the solvent-front, on the dry
part of the paper, with a pencil. See Figure 9.
18.Allow the paper to dry, carefully remove the staples,
and draw an outline around each spot
using a pencil.
19.Take a photograph of your chromatogram after you have circled the spots.
Resize and insert the photograph of your chromatogram in Data Table 1of your
Lab Report Assistant
20.Use the ruler to measure the distance of the solvent front, in millimeters. Use Figure 1 in the
Background section as needed. Record the distance of the solvent front in Data Table 2 of your
Lab Report Assistant
For each of the dye spots on the filter paper chromatogram, measure and record to the
nearest millimeter the distance each of the spots has traveled up the solvent front. Start from
the original horizontal pencil line (base line) and measure to the top center of where the dye
stops in each column on the paper. See Figure 2 in the Background section as needed.
22.Calculate the Rf value for each spot and record in Data Table 2
23.Repeat steps 10 through 22 for the second sheet of filter paper. Record all data for this
chromatogram in Data Table 3 of your Lab Report Assistant
24. Compare the Rf values for the spots of the Kool-Aid ®, unknown sample, store food colorings,and M&Ms ®
, to those of the known FD&C food dyes, to determine which FD&C dyes are used in the Kool-Aid®, the grocery store food colorings, and in the M&Ms ®.
25.Record the FD&C dyes present in each of the samples in Data Table 4 of your Lab ReportAssistant
26.Clean up all equipment and return to the lab kit for future use.
27.When you are finished uploading photos and data into your Lab Report Assistant, save your
file correctly and zip the file you can send it to your instructor as a smaller file.
Enter the results from chromatography filter paper Sheet 1 of your experiment in the following tables. If more than one colour is present in a sample, please indicate colour when referring to distance and Rf measurement.
Sheet 1: FD&C Food Colours
| Colour | Blue | Blue | Red | Red | Yellow | Yellow | Solvent |
| Distance (mm) | |||||||
| Rf | -- |
Sheet 1: Drink Mixes
| Substance | Kool-Aid® | Kool-Aid® Strawberry | Solvent |
| Distance (mm) | |||
| Distance (mm) | -- | ||
| Rf | -- | ||
| Rf | -- |
Enter the results from chromatography filter paper Sheet 2 of your experiment in the following tables. Indicate the colours of M&Ms used in the experiment.
Sheet 2: Store Food Colours
| Substance | Store | Store | Store | Store | Solvent |
| Distance (mm) | |||||
| Distance (mm) | -- | ||||
| Rf | -- | ||||
| Rf | -- |
Sheet 2: Candy Colours and Unknown
| Substance | M&M | M&M | M&M | Unknown | Solvent |
| Colour: | Colour: | Colour: | |||
| Distance (mm) | |||||
| Distance (mm) | -- | ||||
| Distance (mm) | -- | ||||
| Rf | -- | ||||
| Rf | -- | ||||
| Rf | -- |
Answer the following questions based on your experimental results.
Show the calculation for the Rf value for Red40.
What FD&C colour(s) make up the unknown sample?
What FD&C colours make up the food colourings? State the brand used.
What FD&C colours make up the Kool-Aid® drinks and M&Ms®?