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Quantitative analysis of metal ions in

solution using the spectrophotometer
Experiment 5

Introduction
One of the classical challenges facing chemists is the determination of the amount of a given
substance in a sample. This particular branch of Chemistry is known as quantitative analysis
(from quantity, which means amount). Today’s experiment focuses on the use of the
spectrophotometer for quantitative analysis. Certain solutions will absorb light of a specific
wavelength, and the amount of light absorbed by such a solution will depend on the
concentration of the absorbing compound in the solution. The spectrophotometer is designed
to measure the amount of absorption by the sample, and this measurement can be related to
the concentration of the absorbing chemical species in the sample.

In this practical we will determine the concentration of Co2+ and Cu2+ ions in a given solution.
In order to achieve this, we will:
(a) start off by discussing the theory behind the experiment.
(b) determine the appropriate wavelength for analysing Co2+ and Cu2+.
(c) make a series of dilutions, so that we have varying concentrations of Co2+ and Cu2+ and
measure the absorbances of these.
(d) measure the concentrations of Co2+ and Cu2+ in an unknown solution containing a
mixture of these ions.

Theory (Videos 1, 2 and 3)

Solutions display colour due to the absorption of definite wavelengths of visible light by the
solute. For example, a red solution appears red because the blue component of the incident
white light is absorbed by the solution and the remainder (containing mostly the “red”
wavelengths) is transmitted.
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If light of a given wavelength, , is allowed to pass through a solution which absorbs at that
same wavelength, then the intensity of the light emerging from the solution will be less than
the intensity of the incident light. This is carried out in a spectrophotometer, which is
illustrated schematically in Figure 1.

The monochromator lets through light of a single wavelength, with initial intensity I0. This light
passes through the solution in the sample cell and emerges with intensity I where I < I0. The
actual decrease in light intensity depends on the following factors:
(a) Concentration of the absorbing species in the solution.
(b) Path length of the light through the solution. A detector (a photoelectric cell or “electronic
eye”) measures the intensity of the emerging light beam and displays it on a meter.

The relationship between the light intensities I0 and I is given by the Beer-Lambert Law which
can be represented mathematically as follows:

= 10 − cl
I
I0

where
I = the intensity of the light of fixed wavelength transmitted by the sample;
I0 = the light of the same wavelength incident on the sample;
l = the path length of the light through the solution;
c = the concentration of the solution in moldm-3 and
 = a constant characteristic of the absorbing substance at the wavelength of the
incident light.
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= 10 − cl
I
Now if
I0

I0
Then log 10 = cl
I
I0
 A = cl (where A = log 10 )
I
Therefore, the absorbance A is proportional to c, the molar concentration of the solution.

The aims of this experiment are:

(1) to determine the wavelength where the absorbances of Co 2+ and Cu2+ solutions are at a
maximum.
(2) to show that the absorbance of solutions of each of these ions is proportional to the
concentration (at a fixed wavelength and using a fixed path-length).
(3) to analyse a solution which contains a mixture of Co2+ and Cu2+ ions by measuring the
absorbances at two different wavelengths and thereby determining the concentration of
each ion.

Questions

1. Use the Beer-Lambert Law to predict what would happen to the absorbance (A) if the
concentration (c) of a solution doubles.

2. Compound A has absorption maxima at 412 nm, 520 nm and 630 nm, while compound
B has absorption maxima at 420nm, 620 nm and 760 nm. If you are supplied with a
mixed solution of compounds A and B, which wavelengths would you use to determine
the individual concentrations colourimetrically? Concisely explain the reasons for your
choices.
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Procedure

The instructions below have been written for you as if you were actually in the laboratory and
doing the experiment yourself.
For this experiment you will need:
• Approximately 50 cm3 of 0.10 M Co2+ solution, and 50 cm3 of 0.10 M Cu2+ solution in
separate 100 cm3 beakers.
• A sample of the “unknown” solution containing a mixture of Co2+ and Cu2+, in a clean,
dry test tube
All waste from this experiment must be discarded in the “Inorganic Waste” containers.

STEP 1: Determine max for Co2+ and Cu2+ (Video 4)

Using the spectrophotometer, measure the absorbances of 0.10 M solutions of both Co2+ and
Cu2+ ions at 50 nm intervals, starting at 400 nm and ending at 800 nm.
(a) You will find 3 small test tubes or cuvettes in beakers next to the
spectrophotometers; fill one with deionised water, the second with the Co2+
solution, and the third with the Cu2+ solution.
(b) Note the instructions for the use of the spectrophotometer before proceeding with
the experiment.
(c) Set the wavelength at 400 nm. Insert the test tube containing the de-ionised water
and close the lid. Press CAL on the spectrophotometer and wait until the
absorbance is set to 0.000. Determine the absorbance A of each of the other two
solutions at this wavelength.
(d) Determine the absorbance A of both solutions at 50 nm intervals, ending at 800
nm. Be sure to record the absorbances of both solutions at each wavelength.
NB: Remember to zero the spectrophotometer (using the deionised water) at each
new wavelength.
(e) Plot a graph of absorbance (A) vs. wavelength () for each solution with  on the x-
axis. Plot both graphs on the same set of axes. This has been done for you. Refer
to Video 4 and to Slide 7 in the Powerpoint presentation.
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Think about:

(a) At which wavelengths (max) do the absorbances of Co2+ and Cu2+ ions
respectively reach their maxima?
(b) Which respective wavelengths would you recommend for determining the
concentrations of Co2+ and Cu2+ ions?

STEP 2: Prepare samples of varying concentration and read their

absorbances (Videos 5 and 6)
Solutions of known concentration are called standards. Dilute the given 0.10 M solutions
as instructed below:
(a) Use your graduated pipette to prepare four Co2+ solutions (in test tubes) as shown
in the table below. Mark each test tube and mix each solution thoroughly.

Test tube 1 2 3 4
Final concentration 0.025 M 0.050 M 0.075 M 0.10 M
Vol. 0.10 M Co2+/Cu2+ 2.5 cm3 5.0 cm3 7.5 cm3 10 cm3
Vol. Deionised H2O 7.5 cm3 5.0 cm3 2.5 cm3 0 cm3

(b) Repeat step (a) above to prepare four Cu2+ solutions (in test tubes) from the given
0.10 M Cu2+ solution.

Obtain standard (calibration) graphs:

(c) Measure the absorbance of the four Co2+ solutions at the max(Co2+)
determined in Step 1. Measure the absorbance of the four Cu2+ solutions at the
max(Cu2+) you determined in Step 1.
(d) Plot graphs of A (measured at max) vs. c for the two cations being analysed. These
graphs are known as standard or calibration graphs.
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STEP 3: Analyse the “unknown” solution (Video 7)

Analyse the unknown solution containing both Co2+ and Cu2+ ions by measuring its
absorbance first at max(Co2+) and then at max(Cu2+). By referring to the two plots obtained in
Step 2, you will be able to calculate the concentrations of the Co2+ and Cu2+ ions in the
unknown sample.

Report
Submit your report electronically on the report sheet provided in the resources for this
experiment. Please only submit one document. Your report will need to include the
following:

1. Write your answers to the pre-practical questions on page 3 in the spaces provided on
the report sheet.

2. Plot standard graphs of:

Concentration (on the x-axis) vs absorbance (on the y-axis) for both Co2+ and Cu2+.
These may both be plotted on the same axes. They must be plotted on graph paper or
done on the computer using a data analysis programme such as Excel. The graph must
be copied to the report sheet. Alternatively, you may take a photograph of the graph and
insert this into the report sheet.

3. The following results:

(a) [Co2+] in unknown solution = ______ moldm-3
(b) [Cu2+] in unknown solution = ______ moldm-3
Describe how you arrived at these concentrations.