COMPLEX FORMATION: THE METHOD OF CONTINUOUS

VARIATION USING SPECTROPHOTOMETRY

PURPOSE

The purpose of this experiment is to determine the stoichiometry and formation constant for the
equilibrium between Fe
3+
(aq)
and SCN
-
(aq)
illustrated in the reaction:

Fe(H
2
O)
6
3+
(aq)
+n HSCN
(aq)
<==> Fe(H
2
O)
6-n
(SCN)
n
3-n
(aq)
+ n H
3
O
+
(aq)
(1)

where n is the stoichiometric coefficient for the balanced reaction. The stoichiometric ratio and
equilibrium constant will be found using the method of continuous variation.

INTRODUCTION

The iron-containing species in the reaction shown above (see equation 1) are called complex ions. This
class of reactions is often called complexation; it is an important class of reactions, particularly for
transition metal ions. The species that binds to the metal ion (in this case H
2
O and SCN
-
) are called
ligands. Chemical methods of analysis are usually destructive methods. Thus, any attempt to measure the
concentration of either product (or that of the reactants) chemically would destroy the species being
analyzed. This would cause a shift in the equilibrium to compensate for the change creating more (or less)
of the analyzed species. Error in the equilibrium constant would result. However, in many cases complex
ions of transition metals are intensely colored. The color also changes depending upon the nature of the
complexing ligand. The often-intense color of these complex ions is due to their absorption of light from
the visible portion (300 - 700 nm) of the electromagnetic spectrum. This property will be used in the
experiment to monitor the concentration of the iron (III) thiocyanate complex (the product of reaction 1).

In the method of continuous variation, Fe(III) and HSCN solutions are combined so that the mole ratio of
Fe(III)
(aq)
: HSCN
(aq)
varies over a wide range while the total number of moles remains constant. Thus,
the maximum amount of complex will form when the Fe(III)
(aq)
: HSCN
(aq)
ratio is the stoichiometric
ratio. Since the iron and HSCN solutions absorb light weakly in the region of the spectrum where the
complex absorbs strongly, spectrophotometry can be used to determine the relative quantities of all three
species present at equilibrium. This is a nondestructive method of measuring the amount of complex
present and will not disrupt the equilibria. In this experiment the Fe(III)
(aq)
and HSCN
(aq)
stock solutions
both have concentrations of 4.00 x 10
-3
M in a solution that is 0.50 M in HNO
3
. These are combined in
volume ratios of 0.5 : 9.5 through 9.5 : 0.5. As you can clearly see, this will vary the mole ratio of
F(III)
(aq)
and HSCN
(aq)
present in the solution from 0.5 : 9.5 to 9.5 : 0.5. When one of the species is
present in a limiting quantity, the amount of complex that could potentially form is limited. On the other
hand, when each species is present in the correct stoichiometric ratio, the amount of complex that can
potentially form is not limited by either reactant. Also note that the concentration of HNO
3
used is quite
high relative to the Fe(III) and HSCN. This is done intentionally to eliminate changes in pH that would
result due to the production of H
3
O
+
by the equilibria. All of this probably sounds a bit abstract, so lets
examine the way this works by looking at the formation of another complex.

An example of the results of a method of continuous variation experiment for the equilibrium between
Fe(II) and o-phenanthroline (a bidentate ligand) is shown below:

m Fe
2+
(aq)
+n C
12
H
8
N
2

(aq)
<==> Fe
m
(C
12
H
8
N
2
)
n
2m
(aq)


11

10 9 8 7 6 5 4 3 2 1 0
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
Corrected
Metal Moles (relative #)
Absorbance
max. A
observed A

The maximum absorbance occurs at the 2.5 to 7.5 (Fe(II) to o-phenanthroline) ratio. This indicates that
the equilibrium can be written as:

Fe
2+
(aq)
+3 C
12
H
8
N
2

(aq)
<==> Fe(C
12
H
8
N
2
)
3
2+
(aq)


In order to determine the formation constant, K
f
, for this reaction, the concentrations of all three species
present at equilibrium must be known. If the concentration of the original stock solutions of Fe(II) and of
o-phenanthroline were both 2.00 x 10
-4
M, then a 1 : 3 mixture of the two would have [Fe
2+
]
init
=5.0 x
10
-5
M and [C
12
H
8
N
2
]
init
=1.5 x 10
-4
M (you should be able to do the dilution calculation to find these
values).

The extrapolated lines are used to find the absorbance that would occur if the theoretical maximum
amount of complex actually formed (i.e. if the equilibrium constant was very large). From the two initial
concentrations above and the 1 : 3 stoichiometric ratio, it is easy to see that the maximum concentration
of complex that could be formed (all reactant converted to product) is 5.0 x 10
-5
M (you should be able to
determine this number if you do the simple stoichiometric conversions). Beers Law states that
absorbance is directly proportional to concentration. So, from the extrapolated absorbance for the
maximum concentration of complex and the measured absorbance for the complex, the concentration of
the complex actually formed is:

max max
] [
] [
A
obs
A
complex
obs
complex
=
3 2
] ][ [
] [
phen o Fe
complex
K
f

=
+


12
Now the concentrations of all three species at equilibrium can be found and the equilibrium constant can
be calculated using the relationship for K
f
above.

EQUIPMENT

Each pair of students will have access to (a) a Spec 20 and sample tubes, (b) stock solutions of 4.00 x
10
-3
M Fe
3+
(aq)
in 0.50 M HNO
3
and 4.00 x 10
-3
M SCN
-
(aq)
in 0.50 M HNO
3
, and (c) the necessary
glassware for pipeting and diluting solutions.

PROCEDURE

Turn on the Spec 20 and allow it to warm up for approximately 20 minutes prior to making any
measurements. Spectrometers need to have the blank (100% T or 0.000 A) setting reset each time the
wavelength is changed. This is due to three causes: incandescent light sources produce various amounts
of light of different wavelengths, the grating works with an efficiency that varies with wavelength, and
the sensitivity of the detector varies with wavelength. The 0 % T (infinite A) setting should also be
checked each time the wavelength is changed. A solution of a given concentration will absorb light most
strongly at a given wavelength (max). At this wavelength the value of A is most sensitive to changes in
concentration as shown by Beers Law, A =alc (where l is path length of light through the sample, c =
concentration, and a is the absorptivity of the sample which varies with wavelength and has its largest
value at max). You will need to determine max for your complex. You may make your measurements
using the Absorbance or the % T scale (just be sure you know how to reset the blank on the scale you
choose to employ). While the Spec 20 warms up perform the following sample preparation.

Using two burets (one for the Fe
3+
(aq)
and one for the SCN
-
(aq)
) prepare 10.00 mL of solutions that have
ratios of Fe
3+
(aq)
to SCN
-
(aq)
ranging from 0.5 : 9.5 to 9.5 : 0.5 (see the report for the list of solutions to
be prepared). After these are prepared, place each solution into a cuvette. Also, prepare a cuvette
containing the stock solution of Fe
3+
(aq)
and another containing the stock solution of SCN
-
(aq)
. In
addition, you will need a cuvette of distilled water to serve as a blank. You may have fewer than the
fourteen cuvettes you would need for all of the solutions. In this case study the solutions using the
cuvettes you have available taking care to clean and dry the cuvettes when changing solutions.

Inspect your mixtures visually. Which looks the darkest? What might this indicate? Use this solution to
find max for the complex. You will need to measure either A or % T across the entire spectrum
narrowing down the value of max to the single best value (to the nearest 2 nm).

With your spectrometer set and properly adjusted (100 % T or zero A) at max for the complex, measure
either % T or A of all of the cuvettes a minimum of two times. If any two readings seem to be
inconsistent, a third reading is obviously called for. Allow the instructor to examine your data. If the data
is satisfactory then you are done. If not, well you know . . .

REPORT

Complete the attached report form. Your report may be prepared on a computer. The required data
manipulation and graphing should be carried out using a spreadsheet. The spreadsheets and necessary
graphs should be submitted with the report. Make sure to include all data, graphs, and calculations that
impact on the determination you have performed.
13
Prelab Questions that you should think about AND answer prior to coming to lab.


1. Why is a spectrophotometric method used to determine the formula of the complex instead of a
chemical method?




2. Describe in words how the method of continuous variation works. Be sure to explain how it can
be used to determine the formula of a complex.




3. What is the purpose of the extrapolated lines in the Figure on page 36 of your manual? i.e. why
are these lines needed?



4. Be sure you know how to find the concentrations of the Fe
3+
(aq)
and SCN
-
(aq)
present in the
solutions combined in volume ratios of 0.5 : 9.5 through 9.5 : 0.5 (both stock solutions have
concentrations of 4.00 x 10
-3
M). For example, find the initial concentrations present after
mixing:


(a) 2.00 mL of 4.00 x 10
-3
M Fe
3+
(aq)
with 8.00 mL of 4.00 x 10
-3
M SCN
-
(aq)






(b) 7.00 mL of 4.00 x 10
-3
M Fe
3+
(aq)
with 3.00 mL of 4.00 x 10
-3
M SCN
-
(aq)





5. Assume for a moment that the complex had the following stoichiometry:

1 Fe
3+
(aq)
to 4 SCN
-
(aq)


based on your answer to number 4 (a), what is the maximum concentration of the complex
Fe(SCN)
4
-
(aq)
that could be formed in the mixture of 2.00 mL of 4.00 x 10
-3
M Fe
3+
(aq)
with
8.00 mL of 4.00 x 10
-3
M SCN
-
(aq)
?




14