Experiment 7: Spectrophotometric Determination of the Stoichiometry of a Complex

Dela Cruz, Richard Dean Clod C.

Group 4, Chem 27.1, WAD1, Sir Jet Torres
November 29, 2014
I.1Abstract
Spectrophotometry has different methods in which the stoichiometry and composition of an unknown
complex can be determined. In this particular experiment, the stoichiometric ratio of Iron (II) ion and 1,10phenanthroline was investigated by measuring absorbance of a 508 nm UV-vis light source. The different
methods were: the a) continuous variation method, where absorbance is plotted aganst mole fraction, b)
mole-ratio method where absorbance is plotted against mole-ratio and c) slope-ratio method where the two
slopes of two different cases of concentrations are compared with each other. Through these three
methods, the complex was found out to have a 1:3 ratio of moles, which is the theoretical number of moles,
Fe(C12H8N2)32+.
II. Keywords: spectroscopy, complex, mole-ratio, mole fraction, slope ratio, continous variation

III. Introduction
Spectrophotometry is the quantitative
measure of how a material or unknown solution
transmits or reflects certain properties as a
function of wavelength. Spectrophotometric
tiration, is the use of the spectrophotometer as
the indicator in said titration. After observing
multiple instances of concentrations and variation
in the solutions, one measures them in the
spectrophotometer and plots the absorbance
much like in a titration curve. In this experiment
we tested Iron (II) and phenanthroline through
different methods of spectrophotometric titration.
This experiment aims to determine the
stoichiometric ratio of a metal and a ligand in an
unknown complex, and thus also determing its
chemical formula. This is done by three methods:
continuous variation, mole-ratio, and slope-ratio
method.
The continuous variaton methods makes
use of different volumes of the metal and the
ligand are used but the total volume remains
constant. Here, the point where absorbance is the
highest (in the plot of absorbanec vs mole
fraction), is where the stoichiometric ratio lies.
This is because the proper complex formed, has
the highest absorbance.

constant, while the other reactant (usually ligand)

is added in excess. When the graph of
absorbance vs mole-ratio reaches a point where
it plateaus, the point where the plateau started is
the ideal stoichiometric ratio. This is because
absorbance does not increase anymore, since a
complex has already formed and excess ligands
do not contribute to absorbance.
Lastly, the slope-ratio forces the
completion of the reaction of the complex by
adding in both reactants in excess in two
separate cases. One is kept constant while you
add the other in excess, then do the same but
with reversed roles; then use linear regression to
find the slopes of their graphs vs absorbance.
The ratio of these slopes determines the
stoichiometric ratio.

IV. Experimental
A. Continuous Variation
Six 50-mL volumetric flasks were labeled
A to F. In each flask, add the following solutions
in the exact same order: 0.0007 M Iron (II)
solution, 5 mL acetate buffer, 1 mL 0.0007 M
hydroxylamine hydrochloride, and 0.0007 M
phenanthroline solution. These can be
summarized
in
Table
1.1

In mole-ratio the concentration and

volume of one reactant, usually the metal is kept
Chemistry 27.1, Spectrophotometric Determination of the Stoichiometry of a Complex

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Flask
mL Fe(II) soln
mL 1,10phenanthroline

A
0
10

B
2
8

C
4
6

D
6
4

E
8
2

F
10
0

The solutions were then diluted to the

mark and mixed thoroughly. They were left to
stand for 10 minutes so that the reaction may
completely finish and would make the complex
develop the color. The flasks were then
measured for their absorbance at 508 nm. Mole
fraction of each flask was computed and was then
plotted against absorbance.
B. Mole-Ratio Method
Six 50-mL volumetric flasks were labeled
A to F. The following solutions wered added to
each flask. To each flask, add 2 mL 0.0007 M iron
(II) solution, 5 mL acetate buffer, 1 mL
hydroxylamine hydrochloride and 0.0007 M
phenanthroline in various volumes.
A

12

15

Flask

mL Fe(II) soln
mL 1,10phenanthrolin
e

C. Slope-Ratio Method
First part: Label five flasks A to E. In each, add 5
mL Iron (II) solution, 5 mL acetate buffer, 1 mL
hydroxylamine hydrochloride, 0.0007
phenanthroline solution.
D

0.5

1.0

1.5

2.0

2.5

0.7

B
2

C
4

D
6

E
8

F
10

1
0

Continuous Variation

0.5

-0.1 0

0.2

0.4

0.6

0.8

Graph 1.1 Data of varying concentraions and the mole

fraction of Fe vs phenanthroline

B. Mole-Ratio method

Flas
k

Table 1.3 Volume of 0.0007 M phenanthroline to be

added to each flask.

Second part: Label five flasks from A to E. In

each, add: Iron (II) solution, 5 mL acetate buffer,
1 mL hydroxylamine hydrochloride, 5 mL 0.0021
M phenanthroline.

A
0

Table 2.1 Data of varying concentrations of Fe and

phenanthroline

0.1

A. Continous Variation

Dilute to mark and measure absorbance at 508

nm for each flask. Compute for mole ratio of Fe
vs phenanthroline and then plot it against
absorbance.

V. Results

0.3

Measure the absorbance in each case at

508 nm, and then using linear regression,
compute for distinctive slopes for each. The
resulting ratio would be the stoichiometric ratio.

Table 1.2 Volumes of 0.0007 M phenanthroline added to

each flask.

Flask
mL 1,10phenanthroline

Table 1.4 Volume of 0.0007 M Iron (II) solution to be

added to each flask

Table 1.1 Volumes of standard Iron (II) solution and

phenanthroline in each flask

Flask
mL 1,10phenanthroline

Flask
mL Fe(II)
solution

Number of
moles
Phen
Fe
7 x 107

2.1 x
10-6

1.4 x
10-6
1.4 x
10-6

Mole
ratio
(Phe
n/Fe
)
0.5

Absorban
ce

1.5

0.156

Chemistry 27.1, Spectrophotometric Determination of the Stoichiometry of a Complex

0.037

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1.2

3.5 x
10-6
5.6 x
10-6
8.4 x
10-6
1.05 x
10-5

D
E
F

1.4 x
10-6
1.4 x
10-6
1.4 x
10-6
1.4 x
10-6

2.5

0.224

0.325

0.322

7.5

0.320

mL
Fe(II)
0.5
1.0
1.5
2.0
2.5

Flask
A
B
C
D
E

[Fe(II)] in M

Absorbance

7.0 x 10-6
1.4 x 10-5
2.1 x 10-5
2.8 x 10-5
3.5 x 10-5

0.084
0.155
0.224
0.314
0.384

Table 2.3 Data when phenanthroline is constant

Table 2.2 Data of Mole-ratio method of

spectrophotometry (Mole-ratio vs absorbance)

Slope Ratio (phen constant)

Absorbance
0.35

0.5

0.3

0.4

Absorbance

0.25
0.2
0.15
0.1

0.3
y = 10843x + 0.0045
R = 0.998

0.2
0.1

0.05

0
0

10

0.00001

0.00002

0.00003

[Iron(II)] (M)

Graph 1.1 Plot of Absorbance vs Mole-Ratio of complex

Graph 1.3 Plot absorbance vs varying iron

C. Slope-Ratio Method
Flask
A
B
C
D
E

mL
C12H8N2
1
2
3
4
5

[C12H8N2]
in M
1.4 x 10-5
2.8 x 10-5
4.2 x 10-5
5.6 x 10-5
7.0 x 10-5

Absorbance
0.048
0.092
0.142
0.195
0.246

Table 2.3 Data when Iron (II) solution is constant

VI. Discussion
The complex used in the experiment has
the iron ion as the metal, and 1,10 phenanthroline
as the ligand.
Fe2+(aq) + 3C12H8N2(aq) [Fe(C12H8N2)3]2+(aq)

Absorbance

The phenanthroline complex has a

theoretical stoichiometric ratio of 1:3 and has a
deep red-orange color. Specific solutions were
Slope-ratio method (constant
also added to the mixture, to make sure the
Fe(II))
complex is form without any problems. The
acetate buffer was added in order to maintain a
0.3
pH of 2 to 9. Anything higher and/or lower than
y = 3564.3x - 0.0051
R = 0.9989
this range, results to the ferrous ions
0.2
precipitating. The addition of 0.0007 M
0.1
hydroxlamine hydrochloride was added to ensure
that the ferrous ions (Fe2+) dont oxidize into ferric
0
ions (Fe3+). If ferric ion is formed, it will generate
0.00E+00 2.00E-05 4.00E-05 6.00E-05 8.00E-05
a different colored complex with phenanthroline.
[C12H8N2] (M)
Also, 508 nm wavelength was used because this
is the optimum level of wavelength the complex
Graph 1.2 Plot absorbance vs varying C12H8N2
absorbs.
Chemistry 27.1, Spectrophotometric Determination of the Stoichiometry of a Complex
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0.00004

Three different methods were used: (1)

continuous variation, (2) mole-ratio method, and
(3) slope ratio method.

reactant, usually the metal, constant while adding

the other, usually the ligand, in excess. The moleratio is then plotted against absorbance.
Theoretically, same as continuous variation
method, the right stoichiometric ratio of metal to
ligand has the maximum absorbance. So when
the reaction reaches the point where the metal to
ligand mole ratio is right, the graph usually
plateaus. This indicates that maximum
concentration of complex is achieved, and all
other components thereafter do not significantly
contribute absorbance to the solution.

A. Continuous Variation Method

In the continuous variation method, the total
number of moles of the solution were kept
constant, and only mole ratio of each flask varied.
Theoretically, the right metal to ligand ratio of the
complex has the maximum aborbance. It means
that the complex has the highest concentration at
that specific ratio, and no other components
contribute significantly to the solution.

Absorbance

0.7

Continuous Variation

0.5

0.6

0.4
0.5

0.3

0.4

0.2

0.3

0.1

0.2

0
0

0.1
0
0

0.2

0.4

0.6

0.8

Graph 2.1 Continuous Variation mole fraction (X-axis) vs

absorbance (Y-axis)

As seen in graph 2.1, when you

extrapolate the two sections of the graph
descending and ascending, you get the correct
combining ratio of the complex. The intersection
lies on top of around 0.25 mole fraction of Iron (II)
solution. Because iron (II) solution has 0.25 mole
fraction, then phenenthroline has 0.75, and
therefore 0.25/0.75 is equal to 1/3. This indicates
that the metal to ligand ratio is 1:3.
This method is best applied to ligands
with only one complex. If more than one complex
forms, the different peaks of the graphs would be
more than one, and it would be difficult to
determine the right stoichiometric ratio.

B. Mole-Ratio Method
The mole-ratio method determines the
correct stoichiometric ratio by keeping one

1.2

10

15

Graph 2.2 Mole-Ratio - mole ratio (X-axis) vs

absorbance (Y-axis)

As seen in graph 2.2, the graph plateaus

(almost constant) after a certain point. To find the
plateau point, find the intersection of the
extrapolated line of the increasing part of the
graph, and the flat line of the plateau. The point
of intersection is near 3, therefore the metal to
ligand ratio is 1:3.
This particular method is effective for
large complexes where the ligand can
accodomate more metal, like for example 1:3 is
more favorable than 1:1. When this happens, the
graph would look more identical to the
extrapolated lines, suggesting a clearer point of
plateau. This in turn implies that a there is a large
formation constant and the complex is stable.
C. Slope-Ratio Method
Lastly, in this method the complex was forced
into completion by adding excess amounts of
either metal or ligand. When one reactant is in
excess, the concentration of the product is limited
by the other reactant (not excess). This method

Chemistry 27.1, Spectrophotometric Determination of the Stoichiometry of a Complex

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assumes the complex follows the Beer-Lambert

law, and the reaction is complete.
After getting the plot of the excess reactant
vs absorbance for both cases, the slope of each
is then computed through linear regression. And
combined with Beers law, the ratio of the slopes
would be equal to:
/ 1
= =
/ 2
Where m1 is the slope of constant metal, and m2
is the slope of constant ligand. And in that case,
y would be the moles of metal and x would be the
moles of ligand. It would imply a stoichiometric
ratio of y:x.
The slopes of the constant metal vs constant
ligand in graphs 1.3 and 1.4, is 3564.3 and 10843
respectively. Therefore, 10843/3546.3 is around
1:3. Thus, the stoichiometric ratio of metal to
ligand is around 1:3.

VII. Conclusion and Recommendation

Spectrophotometry can be use to
determine the stoichiometric ratio of the metalligand complex. The three methods used in this
experiment each have their own disadvantages
and advatanges, and knowing the best suited one
would achieve more accurate results, and more
efficient methods. Through these methods, the
metal-ligand ratio of Fe(C12H8N2)3 was found out
to be1:3, which is the actual theoretical ratio.
It is recommended that the proper
handling of the spectrophotometer and the
cuvettes must be practiced at all times, to achieve
proper and accurate results. It is also
recommended to make sure that the complex is
formed properly, and to also prevent
inconsistencies, that may affect the results, from
happening.

VIII. References
Harvey, D. (1999). Modern Analytical Chemistry.
USA: McGraw Hill Companies.
Skoog, D., West, D., Holler, F., & Crouch, S.
(2004). Fundamentals of Analytical Chemistry.
Canada: Brooks/Cole-Thomson Learning.
Wear, J.O. (1968). Mathematics of the variation
and mole ratio methods of complex
determination. Retrieved from
http://libinfo.uark.edu/aas/issues/1968v22/v22a1
7.pdf on November 27 2014.

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