Name: _______________

Spectroscopy and Beer’s Law Experiment

Purpose: To determine the relationship between the light transmitted through a solution or
absorbed by a solution with the concentration of copper ions in the solution. A
Spectrophotometer set to a wavelength of 650 nm will be used to measure the
absorbance and transmittance of copper (II) sulphate solutions.
To prepare samples of copper (II) sulphate solutions by dilution.

Theory:
Transition of electrons between quantized energy levels that fall within the visible spectrum is
the basis of visible absorption spectroscopy.

Atomic gases tend to form simple line spectra (Lyman Series for H gas). More complex
molecules, liquids and solids tend to show more broader absorption patterns because of the
interactions (bonding and IMF) with neighboring atoms and molecules. For example, there are
many possible vibrational and rotational energies within a bonded molecule, resulting in no two
molecules showing the exact same energy for an electron transition between the same two energy
states.

http://www.rsc.org/ej/AN/2000/b002215m/b002215m-f1.gif

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Visible Spectroscopy:

Some solutions have distinctive colours, like the deep purple of permanganate ion solutions,
green nickel ion solutions and blue copper (II) ion solutions. A spectrophotometer can measure
the absorption and transmittance of light in these solutions or any solution that absorbs light in
the visible region between 380 to 780 nm. The more absorbing species present, the more light
will be absorbed, and the less will be transmitted through the sample.

Our eyes observe the colour that is not absorbed by a sample. A clear sample absorbs no light so
it transmits all colours, or white light. Our eyes see this as a colourless solution. For coloured
solutions the colour of the sample is the complementary colour of the absorbed EM radiation.
For example, if a sample absorbs yellow, the transmitted wavelengths appear violet. The greater
the concentration of the yellow absorbing substance, the darker the violet colour transmitted.

Colour Wavelengths (nm) Colour

transmitted
Red 750-610 Blue green
Orange 610-595 Blue
Yellow 595-580 Violet
Green 580-500 Purple
Blue 500-435 Orange
Violet 435-380 Yellow

In this lab we will be using spectrophotometers to measure the transmittance of light through
blue coloured solutions with known concentrations to find the relationship between transmitted
light and concentration. Cuvettes, a special optically pure glass with a precise diameter, are
used to hold the sample and one specific wavelength of 650 mn is used for all measurements.

Materials:

Copper II sulphate pentahydrate parafilm beaker

100 mL volumetric flask 0.100 M HNO3* scoopula
Balance weighing paper cuvettes
Spectrophotometer Test tubes pipettes
Parafilm pipette aids
* We use an acid as the solvent (instead of water) to prevent the formation of solid copper salt
precipitates.

Lab Procedure:

You will be working in groups of 2.

• Each group will prepare one 0.200 M copper II sulphate pentahydrate solution in a 100
mL volumetric flask.
• Calculate the mass required for this solution and show your results to the teacher before
you start.
• Weigh out the required mass of copper II sulpate and dissolve it into a volumetric flask
half filled with 0.100 M HNO3. Swirl to mix until no solid is visible.

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 • Fill to line on flask neck, parafilm and invert 10 times to ensure proper mixing.
• Label the flask and poor some of it into a small beaker.
• Calculate the volumes of 0.100 M HNO3 and the 0.200 M solution of copper (II) sulphate
that must be mixed to prepare 5 mL samples of copper (II) sulphate with the following
concentrations: 0.160 M, 0.120 M, 0.080M, and 0.040M
Required Volume of Volume of Total Volume
Concentration 0.100 M HNO3 0.200M CuSO4
0.160M
0.120M
0.080 M
0.040M

• Using the pipettes prepare the four diluted samples in test tubes, seal the tubes with
parafilm and invert to mix.
• Bring the test tubes and the 0.200 M copper (II) sulphate solution to the front bench for
measurement.
• Obtain a cuvette and rinse it with a small amount of the copper II sulphate solution
• Fill the rinsed cuvette ¾ full with the copper II sulphate solution. Do this for each
sample
• All cuvettes must be clean with the outside wiped with lens paper and no air
bubbles in the solution.
The first group to prepare their standard will calibrate the machine
• Adjust the wavelength knob on the spectrophotometer to 650 nm
• Set the instrument to transmission mode and calibrate to 0.0 with no sample in the
sample compartment (or a black card in a cuvette) and to 100% with the blank (0.100M
HNO3) in the sample compartment
• Once the instrument is calibrated it should not be adjusted again so the same settings
are used for all the samples.
• Place your first sample into the machine and measure and record the transmittance.
• Switch to Absorption mode and measure and record the absorbance.
• Repeat for each sample.
• Record your data for all the prepared solutions in the chart below.

Group Data:

Mass of CuSO4•5H2O solid: ________________

Table 1 Class Results of Percent Transmittance and Absorbance of diluted CuSO4 solutions
Solution Molar Measured Measured Absorbance
concentration Percent Transmittance as
Transmittance a Decimal Value
0 0 (Blank) 100% 1.00 0.00
1 0.0400M
2 0.0800M
3 0.120M
4 0.160M
5 0.200M

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Questions:
1. What error would be avoided by using the same cuvette for all the measurements?

2. What error is avoided by rinsing the cuvette with some of the solution before it is filled.

3. Which measured value, transmittance or absorbance, gives a linear relationship with

concentration?

4. Using the concentrations and decimal transmittance values prepare four graphs of:
a) T versus [CuSO4] * b) 1/T versus [CuSO4]
c) logT versus [CuSO4] d) –logT versus [CuSO4]
* [CuSO4] represents the molar concentration of the copper II sulphate solution
Which graph(s) gives a linear relationship? Which one has a positive slope, and passes
through the origin (0,0)? Use linear regression analysis (a line of best fit and r2 value) to
justify your choice (both Excel (c) and Google sheets will do this. Watch the videos Excel:
https://youtu.be/ExfknNCvBYg , for Mac https://www.youtube.com/watch?v=P9BNE0-
6V0Y
google sheets: https://www.youtube.com/watch?v=yZXNS5AxmIY

5. Plot the Absorbance versus [CuSO4]. Which graph from question 4 is the most similar to
this graph? Based on your observations what is the mathematical relationship between the
decimal transmittance and absorbance?

6. What is the equation for Beers Law? Define all variables and explain which are kept
constant.

7. Based on the relationship between transmittance and absorbance if the absorbance is 1.0 what
percentage of light is transmitted to the detector?

8. What would the value be for an absorbance of 2.0 and 3.0?

9. If a 0.10 M solution of a colored substance is measured at a wavelength of 500 nm and an

absorbance of 0.26 is measured, what will be the measured absorbance of a 0.20 M solution
at 500 nm?

Conclusion: