EXPERIMENT #3

Determination of the Mole Ratio of Iron: 1,10-phenanthroline in the Complex

Florin, Neil Anthony L., Ng, Alex Carl L., Solis, Ferguie B.

4-Chem, Department of Chemistry

College of Science, University of Santo Tomas

1. Introduction

Complexation reactions follows this general form: xM + yL MxLy , based on the reaction
of the metal (M) and the ligand (L). These reactions are widely used in analytical chemistry, in the
quantification of the metal ion species using electrochemical oxidation reduction, ion-
chromatography, atomic emission or absorption and measuring the absorbance of a complex
formed between the metal and a complexing agent.

Spectroscopy being a versatile method for complex reactions can record absorbance
measurements without changing the samples nature or the equilibrium of the sample being
measured. As such spectrochemical means of measurement are more common than any other
analytical method. In this experiment, two general approaches, namely Continuous variation and
Mole-ratio method were used.

In the application of continuous variation, the metal cation and the respective ligand in the
solution are both present in identical concentrations but selectively transferred to different
mixtures with the concentration of the total moles being kept constant. As such, the mole ratio is
varying across the set of solutions, furthermore the absorbance measurements for each solution
can be plotted and can then be used to determine the mole fraction using equation 1:

VM/(VM + VL) Eq.1

where VM is the volume for the metal cation and VL is the volume of the ligand.

Lastly, in the mole-ratio method, a series of solutions are utilized with one of the reactants
being held constant while the other is being modified. The absorbance of each solution are
measured and plotted. This method, assumes that the complex absorbs more of the reactants, thus
the plot produced shows less increase for absorbance. This indicates that a break in the slope of
the curves will occur at the mole ratio that corresponds to combining ratio of the complex.

In this experiment the stoichiometry of complex formation for the metal ion Fe2+ and the
complexing agent 1, 10 phenanthroline (otherwise known as phenanthroline C12H8N2) sometimes
referred to as a ligand will be studied. The objective of the experiment is to determine the formula
of the colored complex and using the continuous variation and mole-ratio method and to select the
best method for determining the composition (metal to ligand ratio) of the complex by absorption
measurements.
2. Materials

In this experiment, the reagents used were 0.1M of Acetate buffer prepared at pH 4.0 by
titrating with sodium hydroxide, 0.5M Hydroxylamine hydrochloride, 5x10-4 M 1,10-
phenanthroline solution, 5x10-4 M Fe2+ solution, 1x10-2 M 1,10-phenanthroline solution and 1x10-
4
M Fe2+ solution. The materials used were 100mL volumetric flasks and burette.

For determining the absorbance of each solution, the equipment used was a Lambda 250
UV-Vis spectrometer by Perkin Elmer.

3. Procedure

A. Continuous Variation Method

The group transferred 0,2,4,6,8,10,12,14,16,18 and 20 mL of the Fe2+ solution into 100mL
volumetric flasks, to each flask, 5mL of the acetate buffer were added followed by 2mL of
hydroxylamine chloride. Then, 20,18,16,14,12,10,8,6,4,2 and 0 mL of the phen solution was
transferred respectively to each flask. All of the flasks were diluted to mark using distilled water
and was allowed to stand for 10 minutes.

The absorbance of the solutions prepared was measured using a UV-Vis spectrometer with
water as reference at 508nm. The group then plotted the absorbance of the solutions versus mole
fraction of Fe2+ and calculated for the stoichiometry of the reaction

B. Mole-Ration Method

The group transferred 0.8 mL of the Fe2+ solution using 100mL volumetric flasks, to each
flask 10mL of the acetate buffer were added followed by 4mL of the hydroxylamine solution. Then
0.2,0.6,1.2,1.6,2.8,3.2,3.6 and 4.0 mL of the 1x10-2 M 1,10-phen solution was transferred
respectively to each flask. All of the flasks were diluted to mark and was allowed to stand for 10
minutes.

The absorbance of the solutions prepared was measured using a UV-Vis spectrometer with
water as reference at 510nm. The group then plotted the absorbance of the solutions versus mole
fraction of Fe2+ and calculated for the stoichiometry of the reaction.

4. Results and Discussion

The commonly used methods for determining the stoichiometry of the metal to ligand ratio
spectrophotochemically in a complex are continuous variation and the mole-ratio method. The
absorbance measurements are directly proportional to the theoretical concentration, such that the
concentration of the substance is known to increase. Graphing the absorbance against the
concentration generates a linear line, furthermore in these set of applications, the lines are known
to intersect when extrapolated.
 Continuous variation method is done by using a series of solution in which the
corresponding concentrations of the reactants being used differ in their total molar volume when
added. The mole fraction of the given component or reactant can be calculated using equation 1.

Table 5. Data for Continuous-Variations Method of Determining Stoichiometry of Complex of

Iron(II) and 1,10-phenanthroline
Flask Volume of solution, mL Mole fraction Absorbance,
au
Iron(II) 1,10-phen Iron(II) 1,10-phen
0 0 20 0 1.0 0.032
1 2 18 0.1 0.9 0.064
2 4 16 0.2 0.8 0.096
3 6 14 0.3 0.7 0.101
4 8 12 0.4 0.6 0.091
5 10 10 0.5 0.5 0.081
6 12 8 0.6 0.4 0.060
7 14 6 0.7 0.3 0.052
8 16 4 0.8 0.2 0.046
9 18 2 0.9 0.1 0.036
10 20 0 1.0 0 0.030

The calculated values of mole fraction were then plotted against the absorbance. The
intersection corresponds to the mole fraction of the components that indicates their stoichiometry
as seen on Figure 1. The generated intersection at the where the mole fraction of Iron(II) is
estimated at 0.25 and that of phenanthroline is 0.75. Taking the ratio of Iron(II) with respect to
phenanthroline, the ratio would be 1:3. As such, the maximum absorbance for this complex is said
to occur at the combining ratio of the reactants or the intersection of the extrapolated line.
Furthermore, the complex is said to absorb less than the reactants, a minimum is observed.

0.12

0.1
Absorbance (a.u.)

0.08

0.06

0.04

0.02

0
0 0.2 0.4 0.6 0.8 1
Mole fraction

Figure 1. Plot of mole fraction versus absorbance; for Continuous Variation Method
 In the mole-ratio method, the solutions contain equal amounts of the metal that is being
treated with the increasing amount of the ligand (phenanthroline). The corresponding mole
fractions for this method is found on table 2.

Table 2. Data for Mole-Ratio Method of Determining Stoichiometry of Complex of Iron(II) and
1,10- phenanthroline
Flask Number of moles Mole ratio Absorbance, au
1,10-phen Iron(II) 1,10-phen:
iron(II)
1 0.002 0.008 0.25 0.019
2 0.006 0.008 0.75 0.025
3 0.012 0.008 1.5 0.030
4 0.016 0.008 2.0 0.034
5 0.020 0.008 2.5 0.178
6 0.024 0.008 3.0 0.218
7 0.028 0.008 3.5 0.244
8 0.032 0.008 4.0 0.245
9 0.036 0.008 4.5 0.229
10 0.040 0.008 5.0 0.221

The values when extrapolated forms intersecting line of different slopes. Based on figure
2, it is seen that the intersection of the two lines is at 2.5. The intersection of the extrapolated lines
corresponds to metal ratio; hence the stoichiometry is 1:3. Such that the concentration of the
phenanthroline solution used is thrice the concentration of the iron (II) being used.

0.3

0.25
Absorbance (a.u.)

0.2

0.15

0.1

0.05

0
0 1 2 3 4 5 6
Mole fraction

Figure 2. Plot of mole fraction versus absorbance; for Mole-Ratio Method

5. Conclusion

The commonly used methods for determining the stoichiometry of the complex
spectrophotochemically, namely are continuous variation and mole-ratio method. Based on these
methods, the complex of iron (II) mixed with 1,10-phenanthroline has the metal to ligand ratio of
1:3. Meaning that each molecule of iron present in the solution can coordinately bond three ligands
of phenanthroline.

Furthermore, the mole-ratio method is the best to use such that the complexes having a
large ligand to metal ratio can be distinctly seen on the plot whereas the continuous variation
method, is quite limited to reactants that cannot form more than one complex and is rendered
obsolete when studying complexes with many ligands present.

6. Reference

Atkins, P. W., & De, P. J. (2006). Atkins' Physical chemistry. Oxford: Oxford University
Press

Skoog, D. A., Holler, F. J., & Nieman, T. A. (1998). Principles of instrumental analysis.
Philadelphia: Saunders College Pub