Spectrophotometer Lab
C AP Chemistry
Taher Motiwalla
Introduction:
Spectrophotometers are instruments that measure the amount of light absorbed by molecules in a
solution for a given wavelength. In this exercise you will measure the Percent Transmission
(%T) of a light through a pink, cobalt chloride solution of varying dilutions with water.
According to the Beer-Bouger Law the quantity of light absorbed by a substance dissolved in a
nonabsorbing solvent is directly proportional to the concentration of the substance and the path
length of the light through the solution.
Beer’s law is commonly written in the form A=(epsilon)cl, where A is the absorbance, c is the
concentration in moles per liter, l is the path length in centimeters*, and epsilon is a constant of
proportionality known as the molar extinction coefficient. The law is accurate only for dilute
solutions; deviations from the law occur in concentrated solutions because of interactions
between molecules of the solute, the substance dissolved in the solvent.
The terms percent transmittance and absorbance are used to describe spectrophotometric
analysis. Percent transmittance (T) is the ratio the amount of light passing through the sample to
the amount of light applied to the sample multiplied by 100.:
% T = [(light passing through the sample) / (light shined on sample) ]X 100
Absorbance (A) or Optical Density (O.D.) is directly proportional to concentration of solute and
it represents the logarithm to the base 10 of the reciprocal of the transmittance.
O.D. can be derived from % T according to the following formula:
O.D. = 2 - log10 of % Transmittance
*Note: the path length is given to be equal to 1 cm for the spectrophotometer arrangement in
ChemLab.
Procedure:
Step 1: Obtain a Spectrophotometer from the equipment menu set the wavelength to 510 nm.
Note: the sample for the spectrophotometer is held by a cuvette, which is displayed as part of the
spectrophotometer by ChemLab.
Step 2: Select the Spectrophotometer and add 10 ml of 0.001 M Cobalt Chloride solution from
the Chemicals dialogbox. To insert the cuvette into the spectrophotometer, select it and use the
spectrophotometer insert procedure from the procedures menu (or use right mouse context
menu).
Step 3: Record the Percent transmission from the Spectrophotometer in the observation window.
Remove the cuvette sample from the Spectrophotometer and empty the cuvette.
Step 4: Prepare a diluted solution (1:10) of Cobalt Chloride. Add 10 ml of Cobalt Chloride to a
100 beaker. Then dilute with 90ml of distilled water.
Step 5: Extract 10 ml of the (1:10) diluted solution with the 10 ml pipette and place in
Spectrophotometer’s cuvette and insert the sample in the Spectrophotometer and record the %
Transmission in the observation window. Remove the cuvette sample and empty the cuvette.
Step 6: Prepare a diluted solution (1:100) of Cobalt Chloride. Using a 10 ml pipette extract 10 ml
of the (1: 10) diluted solution from the beaker (step 4). Release the contents from the pipette into
a new 100 ml beaker and dilute with 90 ml of distilled water.
Step 7: Extract 10 ml of the (1:100) diluted solution with the 10 ml pipette and place in
Spectrophotometer’s cuvette and insert the sample in the Spectrophotometer and record the %
Transmission in the observation window.
Observations:
Undiluted solution:
% Transmission of Cobalt Chloride: 31.62%
OD of undiluted solution (OD = 2 - log10 (%T)): 0.5000
Diluted solution (1:10):
% Transmission of Cobalt Chloride: 89.13%
OD of undiluted solution (OD = 2 - log10 (%T)): 0.04998
Diluted solution (1:100):
% Transmission of Cobalt Chloride: 98.86%
OD of undiluted solution (OD = 2 - log10 (%T)): 0.004979
