Analytical Chemistry Laboratory 2

Spectrophotometric Determination of Iron

Mr. *****

Department of Chemical Engineering and Chemistry, Mapua Institute of Technology

A complex of iron (II) in the water sample was formed with 1,10-phenanthroline,
Fe(C12H8N2)32+ and the absorbance of this colored solution is measured with a spectrophotometer.
The volume reading was recorded at 7.0271 mL. The spectrum was plotted to determine the
absorption maximum. Hydroxylamine (as the hydrochloride salt to increase solubility) was added
to reduce any Fe3+ to Fe2+ and to maintain it in that state.

basis for its determination. Since the

Objective solutions are reddish orange, a blue-green

The main purpose of this wavelength of light (508) should be selected

experiment is to determine the amount of
iron in the given sample from reading its
absorbance by using a spectrophotometer. A
series of solutions with different
concentration will be measured to establish a
calibration curve.
Discussion
Overview
The spectrophotometer is a simple
instrument for measuring the amount of
light transmitted through solutions. It has a
diffraction grating that can be used to adjust
the wavelength of light transmitted. In
general the light used should be the
complimentary color of the solution being
tested. The amount of light at a given
wavelength that a sample absorbs is often
directly proportional to the concentration of
the absorbing species in the sample and to
the distance (path length) the light must
travel through the sample.
Iron is a stable ion. It forms an
intense red colored compound with
orthophenanthroline solution, and is the
for the spectrophotometer.
Chemically, iron is an active metal.
It combines with the halogens (fluorine,
chlorine, bromine, iodine, and astatine),
sulfur, phosphorus, carbon, and silicon. It
displaces hydrogen from most dilute acids.
It burns in oxygen to form ferrosoferric
oxide, Fe3O4. When exposed to moist air,
iron becomes corroded, forming a reddish-
brown, flaky, hydrated ferric oxide
commonly known as rust. The formation of
rust is an electrochemical phenomenon in
which the impurities present in iron form an
electrical “couple” with the iron metal. A
small current is set up, water from the
atmosphere providing an electrolytic
solution. Water and soluble electrolytes such
as salt accelerate the reaction. In this process
the iron metal is decomposed and reacts
with oxygen in the air to form rust. The
reaction proceeds faster in those places
where rust accumulates, and the surface of
the metal becomes pitted
Methodology
Preparation of the Calibration Curve
 25 mL of the standard iron solution
was transferred to a 100-mL volumetric
flask, and 25 mL of distilled water to a
second 100-mL volumetric flask. To them, 1
mL of hydroxylamine, 10 mL of sodium
acetate, and 10 mL of orthophenanthroline
were added to each flask. The mixtures were
allowed to stand for 5 min; diluted to the
mark and mixed.
A pair of matched cells for the
instrument was cleaned. Each cell was
rinsed with at least three portions of the
solution it is to contain. The absorbance of
the standard was determined with respect to
the blank.
This procedure was repeated with
five other volumes of the standard iron
solution (1, 5, 15, 35, and 45 mL).
Determination of Iron
10 mL of the unknown was
transferred to a 100-ml volumetric flask;
treated in the exact same way as the
standards and the absorbance was measured
with respect to the blank. The volume of
unknown taken to obtain absorbance
measurements for replicate samples are
within the range of the calibration curve was
altered.
Instrumentation
Spectrophotometer is an instrument
used to measure the intensity of wavelengths
in a spectrum of light compared with the
intensity of light from a standard source.
Interpretation
The red-orange complex that forms
between iron (II) and 1,10-phenanthroline
(orthophenanthroline) is useful in
determining iron in water supplies. The
reagent is a weak base that reacts to form
phenanthrolinium ion, phenH+, in acidic
media. Complex formation with iron is thus
best described by the equation:
Fe2+ + 3phenH+ ↔ Fe(phen)32+ + 3H+
Iron (II) is quantitatively complexed
in the pH range between 3 and 9. A pH of
about 3.5 is ordinarily recommended to
prevent precipitation of iron salts, such as
phosphates. An excess of a reducing reagent,
such as hydroxylamine, was needed to
maintain iron in the +2 oxidation state. The
complex, once formed, is very stable.
The determination was performed
with a spectrophotometer set at 508 nm and
the data were obtained as shown:
V (mL) Absorbance
1 0.0280
5 0.0731
15 0.3478
25 0.4400
35 0.7679
45 0.9901
Unknown 0.1299
The volume of the unknown sample
# 4 was 7.0271 mL, computed by using the
method of linear regression.
Conclusion
Spectophotometers are a vital and
necessary instrument in quantitative
analysis. In this experiment the volume of
iron in the given solution was determined
using this technology. The result obtained
using the calibration curve was 7.0271 mL.
Several reagent were added to the sample to
maintain its stable state throughout the
procedure.
References
1. Christian, G.D., Analytical Chemistry, 6th
edition. New Jersey. John Wiley, 2004.
2. Filby, G. “Spreadsheets for Chemists,”
VCH Publishers, New York, 1995, p. 210.

3. Schwartz, L.M. “Advances in Acid-Base

Gran Plot Technology,” J. Chem. Educ., 64,
947-950 (1987).