edited by

JAMES O. SCHRECK

filtrates & residues University of Northern Colorado

Greeley, CO 80639

Determining Iron Content in Foods by Spectrophotometry

Paul E. Adams • crucible or evaporating dish
Beall High School, E. Main Street, Frostburg, MD 21532 • pipets or burets
Secondary school chemistry students can have • 0.0010 M FeC13 stock solution (at least 11
a satisfying introduction to colorimetry by mL/group) dissolve 0.270 g of FeC13•6H20 in about
obtaining reasonably close results for the 500 ml. of distilled water. Acidify with 5 mL of
determination of iron in various foods. concentrated HCI. Dilute to 1 L with distilled water.
The classic reaction between the iron(lll) ion Mix well. This is more than sufficient. A large quantity
must be made due to the mass of solute required for
and the thiocyanate ion to yield a reddish-brown such a low concentration.
complex ion works quite well for this activity. The
various ions that interfere with the formation of
• 1.5 M KSCN (at least 35—50 mL/group). For
10 groups, dissolve 72.9 g of KSCN in distilled water to
this complex are silver, copper, nickel, cobalt, make 500 ml. of solution. Mix well.
titanium, uranium, molybdenum, mercury, zinc,
cadmium, bismuth, phosphate, arsenate, • 2.0 M HCI (at least 40—70 ml]group). For 10
groups, add 170 ml. of concentrated HCI to 500 ml, of
fluoride, oxalate, tartrate, and sulfate (1). Most distilled water. Mix. Dilute to I L with distilled water.
of these ions either are absent or in low Mix.
concentrations in food. Procedure
ChemCom (2) has a visual comparison activity
related to the iron content in food, and this procedure Part 1. Preparation and Absorbance
can be used as an extension. This experiment works of the Standard Solutions
very well in other chemistry courses as an experience 1. Starting with a 0.0010 M FeC13 stock solution,
in spectrophotometric analysis and its concomitant prepare 10 rnL of each of the following. Use a pipet
skills. It is precisely in this latter context that it is
or buret for the FeC13 volume measurements.
useful.
At high concentrations of thiocyanate ion, the • Caution: Never pipet by mouth. Always use a suction bulb
following equilibrium is pushed far to the right: or similar device for drawing up fluids with a pipet.
3
Use 2.0 M HCI to dilute the stock solution, not distilled
Fe+ (aq) +6 scy (aq) (aq) water. Use a pipet or buret for the HCI volume
The food sample is heated until it is reduced to measurements. Place each solution into a clean dry test
a fine ash. Hydrochloric acid is added to dissolve tube with a minimum capacity of 20 mL. Do not mix with a
the iron in the residue and to prevent the stirring rod because that could contaminate the solutions.
Mix by swirling the solutions in their test tubes. If time is a
hydrolysis of the iron(lll) ion. After filtering, factor, these dilutions will have to be prepared for the
thiocyanate solution is added and the student in advance.
absorbance is determined with a
spectrophotometer. Using a previously prepared a. 0.000050 M iron(lll) ion solution is made by diluting 0.50
iron(lll) complex concentration—absorbance ml. of the iron(lll) chloride stock solution with 9.50 ml, of
2.0 M hydrochloric acid.
graph, the result can be obtained with the help b. 0.00010 M iron(lll) ion solution is made by diluting 1.00 ml.
of interpolation or extrapolation. of the iron(lll) chloride stock solution with 9.00 mL of 2.0
The solution made when the iron(lll) complex is M hydrochloric acid.
mixed with the thiocyanate ion is not stable c. 0.00015 M iron(lll) ion solution is made by diluting 1.50 ml.
under normal temperature and light conditions. Of the iron(lll) chloride stock solution with 8.50 mL of 2.0
The third dilution (described below) lost 5% of its M hydrochloric acid.
absorbance within 10 min and 10% of its d. 0.00020 M iron(lll) ion solution is made by diluting 2.00 ml.
of the iron(lll) chloride stock solution with 8.00 ml. of 2.0
absorbance in 30 min. Spectrophotometric M hydrochloric acid.
readings should be made without delay. e. 0.00025 M iron(lll) ion solution is made by diluting 2.50 ml.
of the iron(lll) chloride stock solution with 7.50 ml, of 2.0
Equipment and Supplies List M hydrochloric acid.
• spectrophotometer—meter or digital 2. Add 5.0 ml. of 1.5 M KSCN to each of the
readout (digital readout is preferred). Colorimetric prepared solutions. Mix each solution well. This will dilute
probes with associated software are available for 10 mL of solution to 15 ml. and cause its concentration to
attachment onto computers (IBM, Mac, Apple Il). decrease to two-thirds its original molarity. This two-thirds
These will gather the data and graph results.
have at least 10—15 ml,
capacity. With a buret or 2.0
pipet, add 10.00 mL of 2.0 M I-
ICI to dissolve the iron(lll)
present in the ash. Stir gently
for about five minutes. Filter
the mixture. Collect the
filtrate.
4. Mix 5.00 ml. of the
filtrate with 5.00 ml. of 1.5 M
KSCN. Mix well. Because the
solution concentration is
halved by adding the 5.00 mL
of KSCN, the concentration of
the filtrate will be twice what
is determined by absorbance.
5. Obtain the absorbance
of this solution soon after
mixing. The color does change 2
significantly within 15—30
min.
6. If the laboratory drain is connected to a municipal waste treatment plant and if local regulations permit,
the solutions ordinarily can be disposed of through dilution.
Absorbance versus concentration of iron thiocyanate ion. Note: the actual concentration, for example at
concentration is the actual concentration that is being Data and Calculations 1, is 0.00010 or 1.0 x 104 M.
read in the spectrophotometer.
3. Read the absorbance of each solution on a Part 1
spectrophotometer set at 480 mg wavelength (I). This step 1. Construct a data table of standard comparison
should not be delayed for an extended period because the information: (sample data consisting of average values
color does fade significantly within 15—30 min. from a class are provided)
Part 2. Determination of the Iron Content in a Food
Sample Standard Comparison Information
4. Determine the total moles of iron in solution
I. Obtain the mass of a clean, dry crucible (or in Part 2, step 3. Determine the mass (in
evaporating dish if the food sample is going to be milligrams) of that many moles of iron. Because
too large). Add about 5 g of a finely chopped sample there were 10 ml, of filtrate:
of the food to be tested and determine its mass to
Molar concentration in filtrate x 0.01000 L = moles of iron
three decimal places.
2. Heat the sample over a Bunsen burner until it has
been reduced to an ash. The heating process could cause
some smells and smoke, so use a well-ventilated room. A
fume hood is desirable. No splashing should occur.
3. After the sample has cooled, transfer the
contents to a small beaker if the crucible or
evaporating dish does not

Volume 72 Number 7 July 1995 649

 [Fe(lll)] from absorbance 650 Journal of Chemical Education
[Fe(lll)] in complex (6.4 x M) x 0.01000 6.4 x 10-7 moles
step I of complex
ion from step 2
ion
0
0.000050 0.000033
0.48
0.00010 0.000067
0.79
0.00015 0.00010
1.12 0.036 mg 4.540 g x mg/g
0.00020 0.00013
1.42
Conclusion

2. Using the data above, plot molar concentration of the 1. From the Handbook of Chemistry and Physics (3)
solution (M) on the x-axis and absorbance (A) on the y- or ChemCom (2) (pp 535—537) or other sources (4),
axis. Draw a straight line representing the best fit for the determine an accepted value for the amount of iron in the
points on the graph. This graph (see figure) will serve as a analyzed food. Accepted values for the iron content of a
means of converting absorbance to molar concentration few foods are presented in the next section.
when calculations from the food source are done. 2. Determine the percent difference between your
value and the value found in a reference book. What are
Part 2 some reasons for the differences between the two values?
1. Construct a data table like the following: (sample data (Reference value—experiment value) x 100/reference value
provided)
Values from different reference sources often are
Type of food: broccoli not the same. This helps the students understand
Mass of food: 4.540 g
Absorbance of solution prepared from the food
that there are many factors from the sample itself
ash: 0.42 (such as growing conditions, soil, variety of that
vegetable, part of the vegetable analyzed, etc.) that
2. Using the graph from Part I, determine the might result in different analytical outcomes.
concentration of the solution in Part 2 step 4. 3. Discuss your body's needs for iron. Research in
For absorbance of 0.42, the molar concentration of the encyclopedias (5, 7) and health texts will provide ample
solution would be about 3.2 x 10-5 M sources of information
3. Determine the molar concentration of iron(lll) in Student responses to this question usually center around
the filtrate obtained in Part 2 step 3. Remember that the the hemoglobin in the blood or the blood's oxygen carrying
concentration of the filtrate will be twice the role and that is satisfactory. However, the instructor might
concentration of the solution due to the doubling of the wish to add that a 70 kg male has about 4—5 g of iron in
volume when the KSCN was added. his body and 60—70% of it (in the forms of hemoglobin,
(3.2 x 10-5 M) = 6.4 x 10-5 M myoglobin, and heme enzymes) is related to the oxy-
moles of Fe x 55.8 g/mol x 1000 mg/g = mg Fe
(6.4 X 10—7 mol) x 55.8 g/mol x 1000 mg/g = 0.036 mg

5. Determine the mass (in milligrams) of iron per

gram of food used.
Milligrams of Fe in sample/ grams of sample

= 7.9
gen carrying role. The remainder is present as iron interpolation or extrapolation will proceed more
bound to protein in compounds such as transferrin, smoothly.
ferritin, and hemosiderin (8). Comparisons of values obtained experimentally
4. For someone of your sex, age, and weight, what is and the values obtained from the 71st edition
the body's daily requirement for iron? (4—7) ofCRC (3) are shown in the table. Thro samples of
tap water were evaporated to dryness and the
Typical Values of Iron Requirements (3)
Category
residue in the beaker was treated as an ash. The
Age (yr) Weight(kg) Iron (mg)
results from the two analyzed samples were quite
similar.
Iron Supplied by Foods and Water
Sample Experimental Value CRC Value mg Fe/g
mg Fe/g
broccoli frozen 7.9 x 10 8.6 x 10
—2
green peas -2
-2 1.6x 10
almonds cabbage
3.4 x 10 -2
pinto beans 3.6 x 10
-2
200 ml tap water 1.2 x 10 -2
400 ml- tap water 4.7 x 10- 0.6 x 10
2(dried) -2
2.6 x 10 3.0 x 10(cooked)
—4

2.1 x 10
Literature Cited
1. Vogel, Arthur. A Iþxtbook of Quantitative Inorganic Analysis; Wiley: New York,
1961, p 786.
2. ChemCom Chemistry in the Community; American Chemical
Society: Kendall/Hunt, Dubuque, IA, 1993, pp 259-260.
3. Handbook of Chemistry and Physics, 71st ed; Lide, David R; CRC, 1990-1991,
pp732 to 7-90.
4. The World Almanac and Book of Facts, Ed. Mark S. Hoffman; Pharos Books,
1992, "Nutrition", pp 254—256.
5. Encyclopedia Americana International Edition; Grolier Inc., 1989, Vol. 20,
"Nutrition", pp 569—572.
6. Colliers Encyclopedia; Macmillan Educational Co., 1989, Vol. 18, "Nutrition", p
31.
7. Encyclopaedia Britannica Macropaedia, Encyclopaedia Britannica Inc., 1981,
Vol. 13, "Nutrition", pp 418—420.
8. Harrow, B.; Mazur, A. Textbook of Biochemistry; W B. Saunders Co.,
Philadelphia, 1962, p 404.

male 15—18 66 12
male 19—24 72 10
female 15-18 55 15
female 19—24 58 15
5. According to your results, what mass of the
vegetable analyzed would you have to consume to get
your daily supply needs? daily requirement of iron (mg)/
(mg of iron per gram of food)
Results
Good results have been obtained from broccoli, peas,
cauliflower, spinach, beans, nuts, and evaporated tap
water (evaporate 200 ml. or more).
When using nuts, a good bit of smoke and soot
are generated. If soot remains on the side of the
crucible or evaporating dish, the ashes should be
transferred to another container before dissolving
with HCI. Although raisins and other dried fruit are
a good source of iron, the incineration process
might be too long to be worth the lab time.
If the graphing is done on a computer or a graphing
calculator, the axes might have to be switched so that
Volume 72 Number 7 July 1995 651