Beer’s Law Lab Introduction: Absorption spectra of chemical species (atoms, molecules, or ions) are generated when a beam

of electromagnetic energy (i.e. light) is passed through a sample, and the chemical species absorbs a portion of the photons of electromagnetic energy passing through the sample. Absorbance is a direct measure of how much light is absorbed by our sample, thus absorbance is proportional to the concentration of the sample. Since light travels at a constant speed, c = 3.0 x 108 m/s, this implies that the absorbance should also be proportional to the path length of the beam through the sample. When path length (l) is measured in centimeters, and the concentration (c) of the absorbing species is measured in Molarity, the proportionality constant is called the Molar Absorptivity or Molar Extinction Coefficient, ε & having units of M-1cm-1, and our proportionality reduced to the Beer-Lambert Law: A = ε x l x c The Beer-Lambert Law allows you to measure the absorbance (A) of a particular sample and to deduce the concentration of the solution from that measurement. In effect, you can measure the concentration of a particular chemical species in a solution as long as you know the species absorbs light of a particular wavelength. Spectrophotometers are instruments that measure the amount of light absorbed by molecules in a solution for a give wavelength. The instrument is able to take white light and separate it into tis constituent colors, allowing the user to examine the absorption of light of individual wavelengths with nearly 1nm resolution.
If Io is the intensity of light entering a solution and It is the intensity of light exiting the solution, then the transmittance, T, of the solution is given as I t/Io. Transmittance is also expressed as a percentage, (It/Io)(100%). Frequently, the absorbance, A, rather than transmittance is used for the amount of light a solution absorbs. Absorbance is defined by the equation A = -log(T) or A = log(Io/It). The absorbance of a solution depends on the quantity of light absorbed by the species in the solution, the wavelength of the light entering the solution, the length of the solution the light has to pass through, and the concentration of the solution. This relationship is known as Beer's law and is expressed mathematically as A = abc. Here “a” is the proportionality constant (molar absorptivity if concentration units are molarity), “b” is the path length of radiation going through the solution, and “c” is the concentration of the solution. In this experiment an absorption spectra curve of absorption versus wavelength will be obtained for a CoCl2·6H2O solution. From the absorption spectra, the wavelength of maximum absorption is determined and a calibration curve relating absorption to concentration for cobaltous nitrate is prepared. Using the calibration curve, the concentration of an unknown cobaltous nitrate solution is determined by measuring its absorption at the selected wavelength.


To determine the wavelength of maximum absorbance of a cobalt (II) chloride solution. wavelength (on the “x” axis). Materials: 0. 2.0 mL of the solution to a clean cuvette. Repeat Steps 6-8. Record the absorbance on the Data Table. Remember that each time you change the wavelength. you must adjust the absorbance to zero using the “blank” and pressing the CAL button. 6. 10. to validate the Beer-Lambert Law. Prepare a “blank” by placing 5.0 mL of deionized water in another cuvette. Use the up and down arrows to select a wavelength of 450 nm. transfer 5. 9. Obtain 100 mL of 0. 4. Remove the blank and replace it with the cuvette containing the CoCl2. Identify the wavelength with 2 Distilled Water Pipet Ruler 100 mL graduated cylinder Observations: . Place the blank in the sample compartment and press the “Cal” button to calibrate the instrument. 5. Close the lid. Draw a smooth curve to fit the experimental points. Graph the absorbance (on the “y” axis) vs.200 M of CoCl2 solution and distilled water in separate beakers. 7. Change the wavelength to 475. 3. Continue to change the wavelength by 25 nm increments and repeat steps 8-10 until you reach 500 nm. Using a pipet and safety bulb.20 M solution of CoCl2 6 cuvettes Stirring rod Spectrum Procedures/Observations: Procedures: 1. 8. Change the Mode to Absorbance with the left and right buttons. 11. and to use the Beer-Lambert Law to determine the concentration of a solution with unknown concentration.

[Co2+]. 14. 18. be sure to note which one you got. 16. (If there is more than one available. 17. Measure and record the absorbance of each of the series of solutions at the wavelength of maximum absorbance (determined in Part I).) Data: Tube 1 Tube 2 Tube 3 Tube 4 Tube 5 3 . Mix the contents of each cuvette using a clean.) Place the solution in a cuvette and place it in the sample compartment of the Spec. dry stirring rod. Follow this point across until you intersect the line (calibration curve). 13. (An alternative method for finding the unknown’s concentration is to read it from the graph that you constructed. Calculate the concentration of each of the solutions based on the dilution factor. then go straight down to the “x” axis to find the concentration.12. Calculate the concentration using Beer’s Law and the data you gathered. Record its absorbance and the unknown number in the Data Table. Find the absorbance value for the unknown on the “y” axis of your graph. Use a ruler to draw the best straight line through the origin and as close as possible to all of the experimental points. Graph absorbance vs. Obtain about 5 mL of a CoCl2 solution of unknown concentration. Prepare a series of CoCl2 standard solutions according to the following table. 15. add the indicated volume of CoCl2 and the indicated volume of deionized water. the maximum absorbance to the nearest multiple of 25 nm and record in the Data Table. Calculate the slope of this line. Using a pipet.

0 2. 4 .Volume of 0. (mL) Volume of water (mL) 5. 1 2 3 4 5 Absorbance Absorbance nm.0 1.0 2.0 3.0 0 4. [Co2+] Use the data above to construct a graph of Absorbance vs.0 4.0 Wavelength of maximum absorption: ____________nm Wavelength 450 nm 460 nm 470 nm 480 nm 490 nm 500 nm 510 nm 520 nm 530 nm 540 nm 550 nm Absorbance as a function of [Co2+] measured at Tube No.0 1.0 3.200 M CoCl2 sol. Draw the best straight line that will go through the origin and as close as possible to all other data points. Concentration of CoCl2.

Unknown # Absorbance Calculations/Error: 5 .

Transmittance: T= I / IO % Transmittance: % T = I / IO x 100 Absorbance: A = .log10 I / IO Conclusion: 6 .

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