Colorimetry
Prior to the lab, please install the PASCO Capstone software from:
https://www.pasco.com/downloads/capstone/pasco-capstone-update/index.cfm using the license key:
18jkm-9v0t7-arq60-pl0dp-n0q3e-o891j
Background
Beer's Law (also known as the Beer-Lambert Law) is an empirical equation in optics relating the absorption
of light to the properties of the material the light is travelling through.
Beer determined that the absorbance of light through a medium has a logarithmic relationship to the
transmittance of light. He also determined that absorbance depends on the concentration of material in the
medium. He put the relationship of absorbance to path length and concentration into a mathematical form as
follows:
𝐼"
log!" = 𝐴 = 𝜀𝑐𝑙
𝐼
Where:
I0 is the intensity of light before passing through the sample
I is the intensity of light after passing through the sample
A is the absorbance of light,
e is the molar absorptivity of the material (the absorbance of a 1 mol.dm-3 solution of the sample in a 1.00 cm
cell at a specific wavelength),
l is the distance that the light travels through the sample (the path length) and
c is the concentration of the solution.
The value of the molar absorptivity, e, varies from one absorbing material to the next, and also with
wavelength for a particular material.
From Beer’s Law it can be seen that there is direct relationship between the absorbance of light through a
substance and the concentration of the substance. In practice, chemists determine an unknown concentration
using a calibration curve. A calibration curve is made by measuring the absorbance of several known
concentrations of a solution. These known concentrations are referred to as standard solutions. The standards
are then plotted as an Absorbance versus Concentration graph. After the calibration curve has been plotted,
the concentration of unknown solutions can be determined by comparison of the absorbance of the unknown
solution with the calibration curve. If the equation of the line for the calibration curve has been determined,
this should be used to calculate the unknown concentration.
[1]
SAFETY: Wear eye protection, Copper nitrate Cu(NO3)2 – IRRITANT.
[1] A. M. a. S. Ghosh, “Development and Validation of UV/ Visible Spectrophotometric Method for the Estimation of Oxcarbazepine in
Bulk and Pharmaceutical Formulations.,” September 2013. [Online]. Available: http://www.rroij.com/open-access/development-and-
validation-of-uv-visible-spectrophotometric-method-for-the-estimation-of-oxcarbazepine-in-bulk-and-pharmaceutical-
.php?aid=34645. [Accessed December 2017].
IB Chemistry
PROCEDURE:
Preparing the solutions
1. Label four clean, dry, test tubes 1 through 4 and place them in the test tube rack. Label a fifth test
tube as “stock” and a sixth test tube as “unknown”. Place these in the test tube rack also.
2. Pipette 2.0, 4.0, 6.0, and 8.0 cm3 of 0.25 mol.dm-3 copper(II) nitrate solution into test tubes 1 through
4, respectively.
3. With a second pipette, deliver 8.0, 6.0, 4.0, and 2.0 cm3 of distilled water into test tubes 1 through 4,
respectively.
4. Thoroughly mix each solution with a stirring rod. Clean and dry the stirring rod between stirrings.
5. Pipette 10.0 cm3 of 0.25 mol.dm-3 copper (II) nitrate solution into the “stock” test tube to use in the
fifth trial.
6. Pipette 10.0 cm3 of the solution of unknown concentration into the test tube you labelled
“unknown”.
Sensor Calibration (removes any signal due to the solvent)
1. Fill the cuvette with distilled water and cap the cuvette tightly. Wipe the cuvette thoroughly with a
tissue to clean off any finger marks.
2. Open the Colorimeter lid, insert the cuvette, and completely close the lid. (Make sure the lid snaps
shut.)
3. Press the Calibration button on the Colorimeter. The light-emitting diode (LED) in the Calibration
button lights up to show that calibration is in progress.
4. Wait for the LED to turn off. Then open the lid and remove the cuvette.
Record Data
1. Set up the software to record your calibration curve using the “keep mode”.
2. Rinse a cuvette twice with approximately 1.0 cm3 of solution from the test tube 1, and then fill the
cuvette with about 6.0 cm3 of the same solution (test tube 1). Cap the cuvette.
Gently tilt the cuvette back and forth to distribute the solute molecules. Do not shake the cuvette or
create air bubbles in the sample.
3. Thoroughly wipe the outside of the cuvette with a tissue to remove finger marks and place the
cuvette in the colorimeter. Close the colorimeter lid.
4. Click the ¢ preview button to begin recording data.
The digits display shows the absorbance. Record the absorbance value when becomes stable by
clicking on the ü keep sample button.
5. Open the Colorimeter lid and remove the cuvette and discard the cuvette contents.
6. Repeat steps 2 to 5 for each of the remaining test tubes.
REPORT:
Calculate the concentration of the solutions, present your results in a suitable table, construct a standard curve
and calculate the concentration of the unknown solution. Process the uncertainties on the concentrations.
