Determining the Concentration

of a Solution: Beers Law

BACKGROUND
The primary objective of this experiment is to determine the concentration of an unknown
nickel (II) sulfate solution. You will be using the Colorimeter shown in Figure 1. In this device,
red light from the LED light source will pass through the solution and strike a photocell. The
NiSO4 solution used in this experiment has a deep green color. A higher concentration of the
colored solution absorbs more light (and transmits less) than a solution of lower concentration.
The Colorimeter monitors the light received by the photocell as either an absorbance or a
percent transmittance value.

Figure 1

Figure 2

You are to use five nickel sulfate solutions of known concentration. Four solutions are dilutions
of the original standard and the standard solution makes the fifth sample. Each solution is
transferred to a small, rectangular cuvette that is placed into the Colorimeter. The amount of light
that penetrates the solution and strikes the photocell is used to compute the absorbance of each
solution. When a graph of absorbance vs. concentration is plotted for the standard solutions, a
direct relationship should result, as shown in Figure 2. The direct relationship between
absorbance and concentration for a solution is known as Beers law.
From your assigned reading
:http://www.shu.ac.uk/schools/sci/chem/tutorials/molspec/beers1.htm

A=bc + (0)
y = mx + b
A is absorbance on the y axes.
is the molar absorbtivity with units of L mol-1 cm-1 (M-1cm-1) related to the intensity of the
color and constant for a particular aqueous compound/ion at a particular wavelength.
b is the path length of the sample in the cuvette in which the sample is contained. A standard
cuvette size is 1cm. The product,b, is the slope of the line

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Experiment 11

c is the concentration of the compound in solution, expressed in mol/L (symbolized by M).

The concentration of an unknown NiSO4 solution is then determined by measuring its absorbance
with the Colorimeter. By locating the absorbance of the unknown on the vertical axis of the
graph, the corresponding concentration can be found on the horizontal axis (follow the arrows in
Figure 2). The concentration of the unknown can also be found using the slope of the Beers law
curve.

OBJECTIVES
In this experiment, you will

Prepare dilutions of a NiSO4 standard solution.

Use a Colorimeter to measure the absorbance value of each standard solution.
Find the relationship between absorbance and concentration of a solution.
Use the results of this experiment to determine the unknown concentration of another
NiSO4 solution.
Examine the difference between single wavelength collection methods and complete
spectrum collection.

PRELAB EXERCISE

Calculate the concentration of the Nickel solutions in the first four test tubes in the student data
table (below) using the equation MintVint=MfinVfin. M is molarity using units mol/L and V is
volume in L or mL as long as the units are consistent.
Trial
number

0.40 M NiSO4
(mL)

Distilled H2O
(mL)

Concentration
(mol/L)

MATERIALS
two graduated 10 mL pipets
pipet pump or pipet bulb
A supply of distilled water
test tube rack
six cuvettes
five test tubes
tissues
stirring rod and Waste beaker.

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Determining the Concentration of a Solution: Beers Law

CHEMICALS AND PRECAUTIONS

0.40 M NiSO4
NiSO4 unknown solution
Wear safety glasses. CAUTION: Be careful not to ingest any NiSO4 solution or spill any on your
skin. Inform your teacher immediately in the event of an accident.

DATA TABLE
Trial

From Prelab -Concentration

(mol/L)

Absorbance

1
2
3
4
5
6

Unknown number ____

Concentration of unknown

mol/L

PROCEDURE
1. Obtain approximately 30 mL of distilled water to use for your dilutions.
2. Label four clean, dry, test tubes 1-4 (the fifth solution is the 0.40 M NiSO4). Pipet 2, 4, 6,
and 8 mL of 0.40 M NiSO4 solution into Test Tubes 1-4, respectively. With a second
pipet, deliver 8, 6, 4, and 2 mL of distilled water into Test Tubes 1-4, respectively.
Thoroughly mix each solution with a stirring rod. Clean and dry the stirring rod between
stirrings. Use the stock 0.40 M NiSO4 for the fifth trial. Volumes for the trials are
summarized below:
Trial
number
1
2
3
4
5

0.40 M NiSO4
(mL)
2
4
6
8
~10

Distilled H2O
(mL)
8
6
4
2
0

3. Connect the Colorimeter to the computer interface. Prepare the computer for data
collection by opening the file 11 Beers Law from the Chemistry with Computers
folder of Logger Pro.
4. You are now ready to calibrate the Colorimeter. Prepare a blank by filling a cuvette 3/4
full with distilled water. To correctly use a Colorimeter cuvette, remember:

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Experiment 11
a.
b.
c.
d.

All cuvettes should be wiped clean and dry on the outside with a tissue.
Handle cuvettes only by the top edge of the ribbed sides.
All solutions should be free of bubbles.
Always position the cuvette with its clear side facing toward the white reference
mark at the top of the cuvette slot on the Colorimeter.

5. Calibrate the Colorimeter.

6. Open the Colorimeter lid.
7. Holding the cuvette by the upper edges, place it in the cuvette slot of the Colorimeter.
Close the lid.
8. Press the < or > button on the Colorimeter to select a wavelength of 635 nm (Red) for
this experiment. Press the CAL button until the red LED begins to flash. Then release the
CAL button. When the LED stops flashing, the calibration is complete. Proceed directly to
Step 7. If your Colorimeter does not have a CAL button, continue with this step to
calibrate your Colorimeter.
9. You are now ready to collect absorbance data for the five standard solutions. Fill five
separate cuvettes about 2/3 full with your five solutions. Click
. Wipe the outside
with a tissue and place it in the Colorimeter. After closing the lid, wait for the absorbance
value displayed on the monitor to stabilize. Then click
type the calculated
concentration in the edit box, and press the ENTER key. The data pair you just collected
should now be plotted on the graph.
10. Repeat the measurements for the remaining solutions recording the correct concentration
values for each step. When you have finished with the fifth solution (0.40 M NiSO4
solution), click
.
11. In your Data and Calculations table, record the absorbance and concentration data pairs
that are displayed in the table.
12. Export your data to Excel. Plot absorbance vs. concentration. Insert trendline and display
both the equation of the line and the R2 value. Save the plot.
13. Obtain the unknown NiSO4 solution and pour into a clean dry cuvette. Read the
absorbance value displayed in the meter. (Important: The reading in the meter is live, so
it is not necessary to click
to read the absorbance value.) When the displayed
absorbance value stabilizes, record its value in Trial 6 of the Data table.
14. Discard the solutions as directed by your instructor. DO NOT LEAVE THE SOLUTION
IN THE COLORIMETER!
15. Collect the complete spectra of each sample as instructed using the spectrophotometer.

CALCULATIONS:
Show all.

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Determining the Concentration of a Solution: Beers Law

CONCLUSIONS:
Produce and print an Excel graph of absorbance vs. concentration, with a regression line and
R2 for your post lab report. Based on this line calculate the concentration of the unknown an
show your calculations and report this value here. Produce a single graph with the full
spectrum of each concentration and indicate the wavelength for which you determined the
Beers Law plot.

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