Spectrophotometric Determination of Iron in

Drinking Water

Courtney Molvig

Lab Partner: Sarah Lusk

CHEM 254 – 01
Courtney Molvig
CHEM 254 – 01
Lab # 8 – Experiment 10

Introduction:
Even though iron is not hazardous to a person’s health, the EPA standards still limit the
amount of iron in the drinking water to less than 0.3 ppm. Too much iron in water that comes
from our faucets can cause stains on bathroom and kitchen furniture (sink, toilet, and bathtub)
and it can stain laundry. Also, too much iron can cause an unpleasant taste in the drinking
water. Iron is actually extremely important for a person’s health. Iron deficiencies can cause a
person to become fatigue and they will be more susceptible to infections. “The most well-known
role that iron plays in human nutrition is in the formation of the protein hemoglobin, which
transports oxygen to all cells of the body. Iron is also used in cellular metabolism and is found in
many of the body’s enzymes” (Garvin).
In this experiment, we first needed to prepare top standards and working standards with
a known iron concentrations. Once these solutions were made and the concentrations were
known, we then used LabQuest with an Ocean Optics Spectrophotometer to measure the
absorbance of the solution. For each standard and working standards, we created them three
times so that we can have three measured values for each of them. Once we had this
information, we graphed the corrected absorbance verse the concentration of the standard. This
gave us a standard calibration curve. We were then able to use the equation of the standard
calibration curve and the measured absorbance of tap water and an unknown to find the iron
concentration of the tap water and unknown. When calculating the iron concentration, we had to
make sure the dilution factor.
For this experiment, we used tap water from the laboratory we were in. I am assuming
that the tap water from the laboratory comes from the same well that the water from surrounding
homes comes from. The tap water from my home smells like iron and leaves a red-brown stain
on the water faucet and in the toilets. From these qualitative observations, I think that there will
be more iron in the tap water from Moscow, but I do not think it will exceed the standards.

Methods:
To begin this experiment, we prepared reagent solutions. The three solutions that we
made were: hydroxylamine hydrochloride solution, ammonium acetate buffer solution, and
phenanthroline solution. All of these solutions did not have to be prepared using a volumetric
glassware or analytical balance, because it only matters that the same solutions were used
throughout the whole experiment. The hydroxylamine hydrochloride solution was prepared by
dissolving approximately four grams of hydroxylamine hydrochloride in approximately forty to
fifty milliliters of DI water. The ammonium acetate buffer solution was prepared by dissolving
approximately sixty grams of ammonium acetate in approximately forty milliliters of DI water and
then about one hundred forty milliliters of glacial acetic acid were added and mixed to it. The
final solution, the phenanthroline solution, was prepared by dissolving about one hundred
milligrams of 1,10-phenanthroline monohydrate in one hundred milliliters of DI water and then
about two drops of concentrated HCl were added to the solution.
 Once the reagent solutions were made, we began preparing the iron top standards,
working standards, and blank. Unlike the reagent solutions, the iron top standard and working
standards have to be prepared using volumetric glassware. One iron top standard, 200 mg/L,
was prepared by diluting 20 mL of a 1000 mg/L iron standard solution in a 100 mL volumetric
flask. We figured out how much of the 1000 mg/L iron standard solution we needed and how
much we had to dilute it to by using the following equation:
𝐶1 𝑉1 = 𝐶2 𝑉2
The other top standard we made was 20 mg/L. Once the two top standards were made, we
made four working solutions, which had concentrations of 8 ppm, 6 ppm, 4 ppm, and 2 ppm.
For the blank, we added approximately 30 mL of DI water to a 50 mL volumetric flask.
We then added 1 mL of the hydroxylamine hydrochloride solution and mixed the solutions
together. We then added 10 mL of the buffer solution and 4 mL of the phenanthroline solution.
Once mixing these together, we diluted to the line with DI water and then mixed again. To
prepare the working standards that were mentioned above, we used our top standard and the
procedure mentioned above, except for this step we adjusted the amount of DI water we added
in order to compensate for the volume of standards. With the blank and each standard, we
measured the absorbance using LabQuest with an Ocean Optics Spectrophotometer,
measuring the blank and then the lowest to highest standard. We made and measured the
blank and the working standards three times.
Once the working standards and blanks were made and measured, we begin preparing
tap water samples and an unknown. For the tap water samples, we add 30 mL of tap water to a
50 mL volumetric flask, add the reagents as listed above, and then fill the flask with DI water
until it reaches the line. We then waiting until color developed. Once the color developed, we
measured the absorbance the same way we did for the blank and working standards and we
repeated it three times as well. We prepared the unknown by adding 10 mL of the unknown to a
50 mL volumetric flask. We then added the reagents and diluted the rest of the way with DI
water. Again, we measured the absorbance and then repeated this three times.
This whole procedure was given to us by Quantitative Analysis Laboratory Experiments,
7th Edition.

Results:
Table 1: Absorbance of Solutions shows the absorbance of the blank, working
solutions, tap water, and unknown for the three trials. From this information, we were able to
calculate the average absorbance and standard deviation. By taking the average absorbance, I
calculated an average corrected absorbance by subtracting the blank average absorbance from
all of the other average absorbance’s.
Table 1:
Absorbance of Solutions
Concentration Absorbance Absorbance Absorbance Average Standard Average
(M) Trial 1 Trial 2 Trial 3 Absorbance Deviation Corrected
Absorbance
0 (Blank) 0.007 0.003 0.001 0.004 0.003 0.000
3.581E-5 0.415 0.429 0.427 0.424 0.008 0.420
(2 mg/L)
7.162E-5 0.749 0.751 0.755 0.752 0.003 0.748
(4 mg/L)

10.74E-5 1.088 1.082 1.086 1.085 0.003 1.081

(6 mg/L)

14.32E-5 1.424 1.428 1.416 1.423 0.006 1.419

(8 mg/L)

Tap Water 0.011 0.007 0.010 0.009 0.002 0.005

Unknown 25 1.290 1.293 1.290 1.291 0.002 1.287

Figure 2:
Average Corrected Absorbance Vs.
Concentration
y = 12807x - 0.0411
R² = 0.9765
2

1.5
Absorbance

0.5

0
0 0.00002 0.00004 0.00006 0.00008 0.0001 0.00012
-0.5
Concentration (M)

Figure 2 shows a graph of average corrected absorbance versus concentration. The

equation for this graph can then be used to calculate the concentration of iron in the tap water
and the given unknown. The error bars shown above have a 5% value.

Table 2 shows the calculated concentrations of the tap water and unknown. In order to
find the concentrations, we used the corrected measured absorbance and plugged that value
into y and solved for x. Once we solved for x, we multiplied that value by the dilution factor to
get the undiluted concentration. We used the least squares method to determine the uncertainty
in the slop, intercept, and iron concentration. There is also the molar absorptivity of
𝐹𝑒(𝑝ℎ𝑒𝑛)2+3 at 510 nm in this graph. To calculate this, I used Beer’s Law, which is A=εbc. I
assumed the path length was 1 cm.
Table 2:
Calculated Concentrations of Tap Water and Unknown
Concentration Concentration Concentration Uncertainty Uncertainty in Intercept
(M) Trial 1 (M) Trial 2 (M) Trial 3 in Slope
Tap Water 6.260E-6 5.739E-6 6.129E-6 ±8830 ±0.101
Uncertainty of ±3.420E-5 ±3.420E-5 ±3.420E-5
Iron
Concentration
for Tap Water
Molar 1118 523 978
absorptivity of
𝑭𝒆(𝒑𝒉𝒆𝒏)𝟐+
𝟑 at
510 nm (M-1cm-1)
Unknown 25 5.181E-4 5.193E-4 5.181E-4
Uncertainty of ±5.275E-5 ±5.282E-5 ±5.275E-5
Iron
Concentration
for Unknown 25
Molar 2482 2482 2482
absorptivity of
𝑭𝒆(𝒑𝒉𝒆𝒏)𝟐+
𝟑 at
510 nm (M-1cm-1)

Discussion:
According to my calculations, the tap water in the laboratory does exceed the EPA
standards. For trial 1, there tap water has a concentration of 6.260E-6 M, which corresponds to
0.3496 mg/L. The EPA standards limit the amount of iron in drinking water to less than 0.3
mg/L. One reason why the limit could be exceeded is because we did not let the water run for a
while before taking the sample. Since the water was sitting in the pipes overnight, the iron could
have accumulated and that is why we could have gotten such a high value. The reason I bring
this up is because the city of Moscow recommends running you water for a little bit so that
minerals that accumulate inside of the pipes have come off and finished clearing from the water.
If you look at the uncertainty of the equation, there is an extremely high uncertainty. This
can be from doing the equation wrong when calculating the concentration or doing the least
squares method incorrectly. To get the values for the uncertainty, I put my data in excel and
used the preprogrammed codes to calculate the values. Since the uncertainty is extremely high,
I do not have any confidence in the data obtained. For the molar absorptivity of 𝐹𝑒(𝑝ℎ𝑒𝑛)2+ 3 at
-1 -1
510 nm for the tap water, the literature value is 11,100 M cm . My values are off by an order of
magnitude. This could be because of error calculating the concentration, but the error obtained
from the absorbance reading also affected this value.
One issue we ran into while completing this experiment is that after we got done
measuring the absorption of our blank and working standard, we began measuring the
absorption of the tap water. We kept getting a negative value for the absorption of the tap water
and it would continue decreasing, so we recalibrated with DI water. This may have caused our
data to be inaccurate and some of the values from the working standard may have been off
before we caught this issue. An advantage of using spectrophotometric determination of iron in
drinking water is that this procedure is straightforward and quit simple, as long as you know
what you are doing. However, with this method all of the Fe3+ needs to be reduced to Fe2+ and if
not enough of a reducing agent is added, your data can be false.
Conclusion:
From the results I obtained, the concentration of iron in the tap water exceeds the EPA
limit. However, according to my uncertainty analysis, I am not confident in my data at all.
Reading lab reports from online and different water quality websites, I do believe that
spectrophotometry is an appropriate tool to measure iron in water. However, until I get a better
understanding of the process and calculations and practice, I would not be confident in the
results I would obtain. Another method of measuring the iron concentration in water is using an
iron-testing strip. The strip would change color once it has been dipped in water for a little bit
and dried. The color would match with the color on the bottle, similar to the pH strip. Each color
is associated with a different concentration in ppm. However, since this method is strictly
qualitative, the data gathered will not be accurate.
In the future, I would like to recalibrate the spectrophotometer more often to ensure that
the data is not being affected from this. I would also like to be more careful when making the
standards. When adding the reagents, we used a graduated cylinder, but I wonder if using the
pipettes will help increase the accuracy of the data. I would also like to run more trials so that
there are more values to average and hopefully a better average would come from it. I would
also like to change how we measured the standards. For the blank, we made three blanks right
off away and measured the absorbance’s of the blank and moved on to the first working
standard. I wonder if our results would be better if we only measured one blank, then the
working standards and tap water. Then, during every new trial we could recalibrate the machine.

Resources:
Garvin, K.S. (2017, October 03). Health Effects of Iron in Drinking Water. Retrieved November
14, 2017.

Attach your report sheet and copies of your pertinent lab notebook pages.