DETERMINATION OF IRON IN WATER

LAB VIS 5 From Juniata College SIM

INTRODUCTION
Spectrophotometers measure the amount of light absorbed by a sample. All colored solutions
absorb visible light and can, therefore, be analyzed with a spectrophotometer. This experiment
uses a spectrophotometer to determine the concentration of iron (II), or Fe+2, ions in an unknown
sample after analyzing the absorption values for a series of iron (II) standards, solutions of
known concentration. Although solutions containing iron (II) ions are colorless, the addition of
ortho-phenanthroline complexes the iron (II) ions, immediately forming an orange species. More
iron (II) ions in a sample result in a deeper orange color that will absorb more light.

PURPOSE
The purpose of this experiment is to determine the concentration of iron (II) in an unknown
sample.

MATERIALS
100 ppm Fe2+ solution
0.25% ortho-phenanthroline solution
graduated 10 mL pipet
10 mL graduated cylinder
5 100 mL volumetric flasks
unknown or collected water sample

Kimwipes
spectrophotometer
computer with Logger Pro
distilled water
cuvette

SAFETY
Always wear goggles and an apron in the lab.

PROCEDURE
Part I: Preparation of standards
1. Label 5 100 mL volumetric flasks: 0 ppm (or blank), 1.0 ppm, 2.0 ppm, 4.0 ppm, 6.0 ppm.
2. Use pipets to deliver 0.0 mL, 1.0 mL, 2.0 mL, 4.0 mL, and 6.0 mL of the 100 ppm Fe+2
solution to the appropriate 100 mL flasks.
3. Add 5 mL of 0.25% ortho-phenanthroline solution, measured in a graduated cylinder, to each
flask. Add enough distilled water to each flask to dilute to the 100.0 mL mark. These are the
standard solutions.
Part II: Setting up the Spectrophotometer

1. Connect the spectrophotometer to the computer using the USB cable. The cuvette holder /
light source should already be connected to the spectrophotometer.
2. Start the Logger Pro program.
3. If the ROY G BIV background does not appear, select the Connect Interface Spectrometer
Scan for Spectrometer from the experiment menu.
4. To calibrate the Spectrometer, chose Calibrate Spectrometer from Experiment menu. The
calibration dialog box will display the message: Waiting60 seconds for lamp to warm
up. The minimum warm up time is one minute. NOTE: For best results, allow
the spectrometer to warm up for at least five minutes. Following the
instructions in the dialog box to complete the calibration, use a rinsed cuvette
filled about full with the 0 ppm Fe2+ solution as instructed. Be sure to
wipe the outside of the cuvette with a Kimwipe and check to make sure
the non-frosted, clear sides are in the light path. The cuvette should be
inserted all the way through the cell holder. You should feel that the
cuvette is gently, but firmly, held in place so that you cannot twist the
cuvette. Click Finish Calibration and then click OK.
Part III: Finding the wavelength of maximum absorbance and setting the collection settings
1. Rinse a cuvette twice with about 1-2 mL of 6 ppm Fe2+ solution. Dispose of the solution as
directed by your instructor.
2. Fill the treated cuvette about full of 6 ppm Fe2+ solution. Clean the cuvette with a Kimwipe
and place sample in the cuvette holder of the spectrometer. Click

Click

to end the data collection.

3. Add a title to your graph by double clicking on the top of the graph and adding title in the
box. Print a copy of this graph for each member of your group.
4. You can read the absorbance using the Examine tool, by clicking on
. Then move the
cursor along the spectrum. The wavelength and absorbance will be displayed in the new
dialog box in the data window. Determine the wavelength of maximum absorbance. Use this
wavelength throughout your experiment.
5. Click on the Configure Spectrometer Data Icon
located on the right hand side of the
toolbar. Under the Set Collection Mode icon in the display click Abs vs. Concentration.
The wavelength of the maximum absorbance will automatically be selected. You may choose
a different wavelength by unclicking the wavelength and clicking another wavelength. Click
Ok when done to close the display. You do not need to store the data.
PART IV: Constructing a Beers Law Graph

1. To set up the graph to reflect ppm, under the data menu, select Column Options >
Concentration. Change the units to ppm.
2. Dispose of the solution in the cuvette. Rinse the cuvette twice with about 1-2 mL of the 1.0
ppm Fe2+ solution. Fill the cuvette full of the 1.0 ppm Fe2+ solution. Wipe and place the
cuvette in the spectrophotometer. Click
. Once the absorbance stabilizes, click
the Keep button. Enter the concentration of the solution and click OK.
3. DO NOT CLICK STOP UNTIL ALL STANDARDS HAVE BEEN RUN! Repeat the above
step to obtain absorbance readings for each of the other known Fe2+ solutions. Click Keep to
save each absorbance reading.
4. After the final known Fe2+ solution, click
5. Click linear fit

to see the equation for the standard solutions.

Part V: Preparation and analysis of the unknown

1. Fill a cuvette about full with your unknown, which will be provided by your instructor.
2. Record the absorbance of the unknown solution in the Data Table.
3. Chose Interpolation Calculator (not Interpolate) from the Analyze menu. A helper box will
appear, displaying the absorbance and concentration of the unknown. Record the concentration
of the unknown. Click Ok.
4. Move the boxes for the linear fit and interpolation calculator so that they are easily readable
and do not interfere with the line. Add a title to the graph. Print a copy of your graph for each
member of your group.

Names_________________________________
Period_________ Class___________________
Date___________

DETERMINATION OF IRON IN WATER

DATA TABLE
Concentration
0.0 ppm (blank)
1.0 ppm
2.0 ppm
4.0 ppm
6.0 ppm
Unknown _____

Absorbance

CALCULATIONS:

Calculate the concentration of iron in the unknown solution in both ppm and M.

QUESTIONS (to be answered in lab report)

1. Why was ortho-phenanthroline added to the solutions?

2. What is the purpose of preparing and analyzing standard iron solutions?

3. What other items could be analyzed using this method?