F Activity 4: Confirmatory Tests

Spectroscopy

P
P R E P A R I N G

WHAT DO YOU THINK? • LE ARNING OBJECTIVES

To prepare for this lab, you have done some reading on

spectroscopy, the study of the interactions of different
wavelengths of light and matter. The equipment being used, a
spectrometer, is an accurate and precise tool that helps limit the
amount of uncertainty in your data. It works by being able
quantitatively and qualitatively emit or absorb a variety of
different wavelengths of light. Because of this, the tests you run
today are confirmatory tests, tests that will identify a substance
to an acceptable level of uncertainty. To make sure you
understood the reading and important background information on
spectroscopy, answer the preparing questions below.

1. What is the relationship between energy of the photon and wavelength?

2. What are the two ways we can observe in spectroscopy?
3. What wavelengths of light are being absorbed by a blue shirt? Reflected?
4. What part of the atom is interacting with light in spectroscopy?

As always, include the objectives or essential questions for this laboratory activity and share these
objectives with a teammate or laboratory partner.

Based on Vernier and Pretzer

1
E
E X P E R I M E N T I N G

EMISSION · ABSORPTION

Part A: Emission Spectroscopy

This first confirmatory test will allow us to confirm the presence of a specific metal ion in a compound.
You will use the qualitative observations (flame color) and quantitative observations (specific wavelengths
of light emitted) to help confirm the presence of a specific metal ion in a mysterious white powder.
You will keep a procedure and observations experimenting section for this part of the lab. Please be sure
to keep details of how you do these, so that you could repeat this if necessary when solving the crime!

1. Use a USB cable to connect a spectrometer to your LabQuest. Connect a SpectroVis Optical
Fiber to the cuvette holder of the SpectroVis.
2. Turn on the LabQuest.
3. To prepare the spectrometer for measuring light emissions, open the Sensors menu and choose
Change Units ► USB: Spectrometer ► Intensity.
4. On the Meter screen, tap Mode. On the Data Collection screen, change the Sample Time to 90
ms and change the Samples to Average to 1.
5. Ignite the lab burner and adjust the flame so that it is mostly a blue color. The flame need not be
too large.
6. Take the wooden stick that is soaking in distilled water and dip it into one of the salt samples.
7. Measure the emission spectrum of a salt solution by reading the steps below first, then trying to
measure the spectrum. Note: This step is easier to do with two people. One person should
carefully hold the fiber optic cable a little bit away from the flame and a second person should
work with the flame, salt and stick.
a. Click “collect.”
b. Hold the fiber optic cable with your fingers very nearly at the tip of the cable. Aim the tip
of the cable toward the flame, and no close than 5-6 cm from the flame. If your fingertips
are warm, the cable is probably too close to the flame. DO NOT MELT THE FIBER
OPTIC CABLE.
c. Put the stick of salt into the flame.
d. If the graph does not show a series of peaks, try another stick and sample.
e. When you achieve a satisfactory graph, click “stop.”
f. Note the flame color and the data peaks in your laboratory note book as observations.
Be sure to identify the metal ion you are running and its peaks.
g. To save your graph, tap the file cabinet icon next to Run 1.
8. Collect data for each known salt (cation) provided. Keep track of your data in your observations
half of your notebook.
9. Identify the unknown based on the flame color and peaks (if provided).
10. Carefully wipe down the tables to remove any salt residue. You do not want a dirty lab bench to
contaminate future results.

Based on Vernier and Pretzer

2
Part B: Absorption Spectroscopy
1. Prepare the blank, each of the five standards, and the unknown for analysis.
a. Rinse an empty cuvette twice with about 1 mL of distilled water.
b. Use the colored wax pencil to write a zero on the lid of the cuvette.
c. Fill the cuvette 3/4 full with distilled water using a disposable pipette. Seal the cuvette with the lid.
Dry the outside of the cuvette with a tissue.
d. Repeat Steps 1a–1c, using the standard solutions and the unknown, rather than distilled water,
and labeling the lids of the cuvettes appropriately with numbers and “U” for the unknown.

Remember the following:

• All cuvettes should be clean and dry on the outside.
• Handle a cuvette only by the top edge or ribbed sides, not the transparent sides.
• All solutions should be free of bubbles.
• Label the lid of the cuvette so the label does not interfere with the beam of light.

2. Connect the Spectrophotometer to LabQuest and calibrate.

a. Insert the distilled water cuvette into the SpectroVis.

b. Tap on Sensors  Calibrate  USB: Spectrometer
c. Allow the SpectroVis to warm up and then tap Finish Calibration
d. When it reads “Calibration Completed,” select OK.

3. You are now ready to collect absorbance spectra for the various samples.
a. Replace the distilled water with one of the samples to be tested.
b. Click Collect (the play button on the LabQuest machine) or green triangle on the bottom right-
hand corner).
c. Once it the graph has stopped jumping, click the play button again to stop the collection.
d. In your experimenting’s observation section, note the major wavelength peaks. You can find the
wavelengths by clicking on them and reading the value presented in the bottom right-hand corner.
e. Save the graph on the LabQuest, using your group initials and the name of the ink. You may
want to refer back to it when identifying the unknown. You should have them all saved in the
same file but as different runs (there is a file folder in the right-hand corner to go back to different
runs – you used it in Part A, too).
f. Run each of the other standards supplied to you, noting the absorbance peaks in each spectrum
in your laboratory notebook.

4. Run the unknown spectra. Note the letter or number of the unknown and the peaks in your laboratory
notebook. Identify based on the peaks.

1. Take the saved files and transfer them to your computer using the SD card and SD card slot in your
computer. Open these graphs using “Logger Pro” software on your laptop and edit them to make a
clear graph for you to use in your forensics challenge.

a. Select “Data Browser” from the “Data” menu.

b. Double click the current run and name them the type of ink if not already named.

c. You can double click on “Abs” for each run and the Choose “Options” and change the color of
each line (so it is clear in the Legend).

Based on Vernier and Pretzer

3
A
A N A L Y Z I N G

RE ADING • QUESTIONS

Reading: Confirmatory Tests

You did your reading ahead of time for this lab. You may want to go back and reread now that you’ve
had the laboratory experience.

Questions: Confirmatory Tests

1. Take a look at your data from part A. What would you say is a distinguishing peak or two for
each salt? How do those peaks relate to the color of the flame?
2. Take a look at your spectrum of strontium. Calculate the energy associated with the major peaks
ℎ𝑐𝑐
in your data using the formula 𝐸𝐸 = . Note that 𝑐𝑐 = 3.00 𝑥𝑥 108 m/s and ℎ = 6.63 𝑥𝑥 10−34 J∙sec.
𝜆𝜆
Watch your units very carefully, as the wavelength needs to be in meters! You’ll need all your
stellar problem solving skills (unit analysis, sig figs, algebra, exponents) to do this well.
3. What are the general wavelengths in nm associated with the following colors?
a. Red
b. Orange
c. Yellow
d. Green
e. Blue
f. Violet
4. What is the color of light that has the highest energy? How do you know?
5. In part B, why did the inks show different absorbance patterns if they all appeared to be the same
color?
6. Do you think you would have seen the same large variations in absorbance if all the samples had
been red ink or all the samples had been blue ink instead of black? Why or why not?
7. What is the unknown ink (please state the letter associated with your unknown)? How do you
know (justify with your evidence)?
8. Why is this a comfirmatory test and not a presumptive test? Support your response using what
you learned about these terms from your supplementary reading from Crime Scene Chemistry…
and AC3: Presumptive Blood Testing.
9. Compare and contrast absorption and emission spectrophotometry at the macro and nanoscales.

No Critical Thinking for this lab, as the analyzing has a lot of the aspects of How do I know, Why do I
believe, and What does it mean in the questions. You should go back through your laboratory write up to
make sure you can repreat both procedures for an unknown in the lab practical at the end of this unit.

Based on Vernier and Pretzer

4