Experiment 3 – Beer’s Law & Quantum Dots Chem 128

Introduction:

In class, you’ve learned about how the absorption of light is related to quantized energy levels and
that for some systems this can be modeled as a particle-in-a-box. In Part A of this experiment, we
will learn about the instrument and methodology used to measure light absorption in the lab. Then
in Part B, you will synthesize quantum dots of different sizes to observe how the size of a box
affects the energy levels of the particle inside.

To determine the optical properties of the solutions you have prepared, and the quantum dots you
will produce, we will be using UV-visible linear absorption spectroscopy with the MeasureNet®
system on your lab bench. The UV/Vis system works by sending light through your sample and
collecting what passes through the sample. When a molecule in the sample solution absorbs or
scatters some of the light passing through the cuvette the intensity of the light is decreased. The
ratio of the reduced intensity (I) to the initial intensity (I0) of the light is known as the transmittance
(I/I0). However, transmittance can span many orders of magnitude, so it is easier to work with the
log of the transmittance, which is called the absorbance (A). %T is related to absorbance by A = -
log T. A 50% transmittance then corresponds to an absorbance of -log(0.50) = 0.301.

𝐼𝐼
𝐴𝐴 = − log(𝑇𝑇) = − log( )
𝐼𝐼0

The MeasureNet® system works by using a photodiode array spectrometer, where the light that
passes through your sample gets dispersed, and the resulting wavelengths projected onto different
positions along the photodiode array (see Figure 1). The array is calibrated in which each
wavelength will hit at a specific spot, and all photodiodes in the array can be measured
simultaneously. This allows for quick scans of your sample, with 1 nm resolution. For the quantum
dots, you will be collecting a full absorbance spectrum using a wide range of wavelengths while
for your potassium permanganate solutions, you will be detecting at a single wavelength, 520 nm.

Figure 1: A schematic diagram of a simple linear absorption spectrophotometer similar to the MeasureNet system
you will be using for the Beer’s Law and Quantum Dot experiments. The source emits white light containing a wide
range of visible which passes through the sample. The light that passes through the sample is dispersed into
individual wavelength components which are detected at different positions on the photodiode array.

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Experiment 3 – Beer’s Law & Quantum Dots Chem 128

PART A:

Light is a form of electromagnetic radiation. For the purposes of this lab it is best viewed as a form
of energy existing as a wave that can be characterized by frequency and wavelength. Different
wavelengths of visible light correspond to different colors. Light with a wavelength of about 700
nm appears red to us while light with a wavelength of about 400 nm appears violet. The sun floods
the earth with visible light of all wavelengths (white light). Objects have different colors because
different chemical substances absorb different wavelengths of light. Grape juice, for example, does
not absorb purple light; it absorbs purple’s complementary color from the white light but transmits
enough of the purple light that the solution appears purple to us. If we diluted some of the grape
juice with water, we would notice that the purple color is less intense than before. The
concentration of the chemicals that absorb light is smaller in the dilute solution than in the more
concentrated one, so the dilute solution filters out less of purple’s complimentary color.

The Beer-Lambert law explains this effect in a quantitative way. It says that if we pass light of a
particular wavelength through a solution, the amount of light absorbed by the solution is
proportional to the number of molecules present in the solution that can absorb the light.
Mathematically, Beer’s Law is given by the equation

𝐴𝐴 = 𝜖𝜖𝜖𝜖𝜖𝜖

Here, A is the absorbance; 𝜖𝜖 is a constant called the molar extinction coefficient (sometimes called
the molar absorptivity constant) in L/mol∙cm; 𝑙𝑙 is the path length, in cm, of the cell containing the
solution; and 𝑐𝑐 is the concentration of the solution in mol/L. There is a linear relationship between
absorbance and concentration. A more detailed discussion of Beer’s Law including an example
using it that will be similar to the experiment you are conducing can be found in Appendix 3 of
your book (pg. A16)

There are deviations from Beer’s law at extremely high and low concentrations, but in this
experiment, you will be working in the so-called linear dynamic range, where Beer’s Law is valid.
Your objective this week is to demonstrate that this relationship is indeed linear.

Supplies:
• 100 mL volumetric flask
• 2 mL pipet
• 5 mL pipet
• 10 mL pipet
• 50 mL pipet / graduated cylinder
• Cuvette

Chemicals:
• Potassium permanganate (KMNO4)

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Experiment 3 – Beer’s Law & Quantum Dots Chem 128

Hazards and Waste Disposal:

• Potassium permanganate (KMnO4) is a strong oxidizer, be careful with handling and wash
hands after use. May stain skin – will stain clothing!
• All of the waste goes in the heavy metals waste container.

Procedure:
1. Mass 0.1000g of KMnO4, and set aside.
2. Take one 100 mL volumetric flask. Mass the empty flask and record in your notebook.
3. Add the KMnO4 from step 1, and re-mass the flask with the KMnO4 inside. Calculate the
difference in mass, to find the actual amount added, in your lab notebook.
4. Add a small amount DI water, and swirl flask until all solid is dissolved. Add DI water up
to the 100 mL mark on the flask.
5. Mix thoroughly and pour into a clean 150 mL beaker. This is now your stock solution.
(Hint: It would be a good idea to label these!)
6. Rinse volumetric flask well, as you will be using the same one throughout the lab.
7. Take 5.00 mL of the stock solution, and place into the cleaned 100 mL volumetric flask,
fill to the line with DI water, mix, and place in a clean 150 mL beaker. This is now solution
#1.
8. Take 2.00 mL of the stock solution, place into cleaned 100 mL volumetric flask, fill to the
line with DI water, mix, and place in a clean 150 mL beaker. This is now solution #2.
9. Take 50 mL of solution #1, place into cleaned 100 mL volumetric flask, fill to the line with
DI water, mix, and place in a clean 150 mL beaker. This is now solution #3.
10. Take 50 mL of solution #3, place into a cleaned 100 mL volumetric flask, fill to the line
with DI water, and mix. You will not be making any more solutions so this one is fine to
leave in the flask. This is now solution #4.

Now you are ready to use the MeasureNet® to measure absorbance. The MeasureNet® system is
connected to an electronic device that can measure the absorbance of light as it passes through a
solution. We can then scan our sample, observe at a specific wavelength, and read the absorbance.

To get started:

1. Turn on your MeasureNet® at your station and select Spectroscopy.

2. Select ABSORPTION (1) or ABSORPTION (2), remember which you select, as this
determines which spectrometer you will be using. Also remember your station number –
you will need it when at the spectrometer.
3. Have one partner stay at your station, while the other goes to the spectroscopy station.

At the spectroscopy station:

1. The MeasureNet® will ask your station number, enter your station number and press
ENTER.
2. The screen will show ‘Read to scan’ once it communicates with your station at the bench.
3. If you are the first group to use the spectrometer, you will need to calibrate the spectrometer
a. Using the dark zeroing cuvette, insert into the holder and press ZERO.
b. Using a blank cuvette with just water, place in the holder and press REFERENCE.

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Experiment 3 – Beer’s Law & Quantum Dots Chem 128

4. You are now ready to scan your solutions.

5. Fill the cuvette about 3/4ths full of your solution, and place in the holder. Press SAMPLE.
After the scan, data will be displayed on your station at the bench. Have your partner press
FILE OPTIONS on the MeasureNet® and name your samples in sequential order (001, 002,
003, etc.). You must remember to save your file after each scan!!
6. Data files will be sent to you from your TA after the lab and can be read from your software
of choice. For this lab, we are looking for the absorbance at 520 nm!

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Experiment 3 – Beer’s Law & Quantum Dots Chem 128

PART B:

Quantum dots (QDs) are semiconducting nano-materials that can absorb and emit light. These
materials are an excellent example of quantum confinement – when particles are restricted to really
small ‘boxes’, their energy levels become discrete, allowing the absorption of only specific
quantity of energy. In the case of QDs, the particles are delocalized electrons and holes from the
semiconductor, and the ‘box’ is the QD nanocrystal. The energy levels of the electrons in the QD
can be mathematically described using the particle-in-a-box model discussed in quantum
chemistry. As the size of the QD (the box) increases, the spacing between the discrete energy levels
decreases (see Figure 2). Thus, by controlling the size of the box, one can select how much energy
(and correspondingly, the wavelength of light) that the QD will absorb and emit. This allows one
to select the color of the QD. Due to these unique properties, QDs have found broad applications
in biological imaging, color displays, and chemical sensors.

Figure 2. (Left) Schematic energy diagram of QDs. Based on the particle-in-box model, the energy
gap decreases as the size of QDs increases. (Right) Experimental values of the absorption wavelength
and the energy gap of CdSe QDs of various sizes. [Reference: Jasieniak J.; Smith L.; van Embden, J.;
Mulvaney, P. Re-examination of the Size-Dependent Absorption Properties of CdSe Quantum Dots. J.
Phys. Chem. C 2009, 113, 19468-19474.]

The nanocrystal studied in this laboratory is comprised of cadmium (Cd) and selenium (Se)
semiconducting materials. The most typical shape of nanocrystals is spherical, which gives the
name ‘dots’. In Figure 3, an image generated by Transmission Electron Microscope (TEM) shows
an example of the spherical CdSe QDs. In this laboratory you will synthesize CdSe QDs and
examine their optical properties. The preparation involves simultaneously injecting a solution of
selenium and a solution of cadmium acetate dihydrate into an octadecene growth solution
consisting of a small amount of oleylamine. As soon as the Cd and Se solutions are combined,
they begin to form QDs that grow with time. While the QDs are growing, you will remove samples
from solution at various times to obtain a series of QDs of varying sizes. You will then measure
the absorption and emission spectra of each sample. It should be noted that at the end of reaction,
QDs are often covered by organic compounds present in synthesis solutions, as illustrated in Figure
3.

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Experiment 3 – Beer’s Law & Quantum Dots Chem 128

Figure 3. (Left) Sketch of a CdSe quantum dot covered with surfactants: oleic acid (RCOOH),
trioctylphosphine (TOP), and oleylamine (R-NH2). (Right) TEM image of spherical CdSe QDs.

Supplies:

Materials:
• 10 mL graduated cylinder
• 25 mL round bottom flask
• 1 mL glass syringe
• 2 – 1 mL glass pipets
• Pipet pump
• Stir bar
• Heating mantle/stir plate
• Thermometer
• 10 small glass vials with caps
• Pasteur pipets w/ bulb
• 1-cm cuvette

Chemicals:
• 1-Octadecene
• Oleylamine

Hazards and Waste Disposal:

• Octadecene and oleic acid vapors should not be inhaled or in contact with skin and
eyes.
• Trioctylphosphine causes skin burns and eye damage; it should be handled
exclusively with a syringe.
• Metallic selenium is toxic if inhaled, may be harmful if swallowed or absorbed
through skin.
• Cadmium acetate dihydrate is toxic if inhaled or swallowed and is known to be
carcinogenic to humans.
• All waste goes in heavy metals waste container.

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Experiment 3 – Beer’s Law & Quantum Dots Chem 128

Procedure:

The stock solutions will be prepared by your TA and will be heating prior to starting your lab.
They are located in the fume hoods. You will be preparing the growth solution, adding the
precursors, and taking aliquots every few seconds to obtain different sized quantum dots with
different properties.

1. Securely clamp a 3-port 25 mL round bottom flask to a

ring stand using the clamps in your drawer.
2. Measure out 10 mL of octadecene using a graduated
cylinder and place in the round bottom flask.
3. Using a syringe, measure 0.67 mL of oleylamine and add
it to the growth solution.
4. Turn on the stirring, and set to a speed that stirs, but not
splatters the solution.
5. Insert the thermometer in the solution such that it is
adequately submerged in the solution and does not interrupt the stirring.
6. Heat the solution to 165ºC using the heating mantle.
7. While heating, label the glass vials 1-10.
8. Once the temperature of the growth solution has stabilized to 165ºC, use the glass 1 mL
pipet to withdraw 1 mL of the Se stock solution.
9. Use another glass 1 mL pipet to withdraw 1 mL of the Cd stock solution.
*withdraw the Cd solution quickly to avoid its cooling* CAUTION: The Cd solution is
hot and should be handled carefully!
10. Simultaneously inject the 1 mL of the Se solution and the 1 mL of the Cd solution, into
your growth solution.
*start timing as soon as you inject the solutions, the QDs begin forming and growing
instantly when the solutions are combined*
11. Withdraw ~1 mL of the reaction mixture at the times listed in the table below using a
Pasteur pipette, and place in the labeled glass vial.
12. Once samples are all withdrawn, turn off the heating mantle.

Vial # Suggested Actual Withdrawal Color of Sample under Ambient Light

Withdrawal Time Time
1
2
3
4
5
6
7
8
9
10

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Experiment 3 – Beer’s Law & Quantum Dots Chem 128

Procedure:

4. Turn on your MeasureNet® at your station and select Spectroscopy.

5. Select ABSORPTION (1) or ABSORPTION (2), remember which you select, as this
determines which spectrometer you will be using. Also remember your station number –
you will need it when at the spectrometer.
6. Have one partner stay at your station, while the other goes to the spectroscopy station.

At the spectroscopy station:

7. The MeasureNet® will ask your station number, enter your station number and press
ENTER.
8. The screen will show ‘Read to scan’ once it communicates with your station at the bench.
9. If you are the first group to use the spectrometer, you will need to calibrate the spectrometer
a. Using the dark zeroing cuvette, insert into the holder and press ZERO.
b. Using a blank cuvette with just octadecene, place in the holder and press
REFERENCE.
10. You are now ready to scan your QDs.
11. Place ~ 1 mL of your sample in the cuvette and place in the holder. Press SAMPLE. After
the scan, data will be displayed on your station at the bench. Have your partner press FILE
OPTIONS on the MeasureNet® and name your samples in sequential order (001, 002, 003,
etc.). You must remember to save your file after each scan!!
12. Data files will be sent to you from your TA after the lab and can be plotted in your software
of choice (Excel, MATLAB, Origin, etc.).