Establishing an Equilibrium Constant through Spectrophotometric

Analysis of Thiocyanatoiron (II) Ion Complex Solution

Gilbert L. Huizar, Brittany Helaire, Rhonda Shuler
St. Philips College, San Antonio, Texas

Abstract
Spectrophotometric analysis of a series of standardized thiocyanatoiron (II) complex
solutions (of known molar concentrations) resulting in Beer Lambert calibration curve
with a coefficient of determination equal to 0.9957. Absorbance data from
spectrophotometric analysis of five samples, with expected similar equilibrium
concentrations of Iron (III) ion, Thiocyanate ion and Thiocyanatoiron (II) complex, are
correlated with calibration curve equation and established an average equilibrium
constant (Kc) of 309.30 23.08. The experimentally determined average equilibrium
constant resulted in 13.9% error from expected Kc value (at similar conditions) of
271.49.
Introduction
Electromagnetic radiation is a type of energy that encompasses both wave properties and
particle properties. Particle properties of electromagnetic radiation allow plants to excite electrons in
cell membranes and enable light energy to be transferred for photosynthesis occur (3). The wave
property of electromagnetic radiation resembles a rollercoaster, with equally proportional rises and
dips, but with the ability to spin on a central axis. (like a spinning drill bit). The steepness of the rises
and dips is the amplitude while the wavelength is the distance from peak to peak. All electromagnetic
radiation travels at the same speed (3.00 x 108 m s-1), but exhibit differing intensities. These
intensities are determined by the wavelength and the amplitude of the Electromagnetic Radiation (EMR)
(2).
The entire range of EMR is called the electromagnetic spectrum. White light (or visible light) is a
very small fraction of the electromagnetic spectrum but is extremely useful in the field of scientific
research. As stated before, the intensity of the EMR is measured by the wavelength and amplitude; and

specifically, wavelengths between 400 nm and 750 nm are what make up visible light. The color
perceived by a human eye is actually the EMR that is being reflected by a specific substance or matter.
EMR at a specified wavelength can be used to determine how much of that designated
wavelength is reflected or transmitted through a substance. A spectrophotometer is an instrument that
gives scientists the capability to measure the transmittance of EMR through a substance (5). A
spectrophotometer is equipped with a white light source, a prism, control slit and an EMR detector. The
white light passes through the prism, and emits all the colors of the visible spectrum. The slit allows the
specified EMR wavelength to pass through a cuvette and the EMR detector measures the transmittance
of the light. The amount of specified EMR wavelength received by the detector is converted to a digital
readout on the spectrophotometer and is delivered as percent transmittance or absorbance value (5).
Absorption of a specified wavelength can be used to correlate the molar concentration of a
substance that reflects a certain wavelength. This is known as The Beer Lambert Law: The molar
absorptivity coefficient () [a constant value (L mol-1 cm-1) at any specific wavelength for a light
absorbing substance] is multiplied by the path length of the EMR (b) and multiplied by the molar
concentration of the compound in a solution (c) (4). ( Equation 1 )

(1)

When a chemical reaction occurs, a gas may evolve, a precipitate may form and/or a color may
become visible. In the reaction between Iron (III) ion solution (Fe3+) and Sodium Thiocyanate (NaSCN), a
blood red complex becomes visible (1). This blood red color is Thiocyanatoiron (III) ion complex. Using
the principles of the Beer Lambert Law, the concentration thiocyanatoiron (III) ion complex can be
determined by the absorbance data from a spectrophotometer.

A set of calibrated solutions are used to correlate absorbance values (determined by

spectrophotometric analysis) with known molar concentrations of Thiocyanatoiron (II) Ion Complex (1).
By using the stoichiometric equation (Equation 2), for every one mole of Thiocyanate ion (SCN-) present,
one mole of Thiocyanatoiron Ion complex (FeNCS2+) can be produced:

Fe3+ (aq) + SCN- (aq)

FeNCS2+

(2)

Because it is known that the amount of moles of SCN- reacted will equal the number of moles FeNCS2+
produced at equilibrium, the molar concentration of Thiocyanatoiron (III) complex at equilibrium can be
determined by knowing the amount of SCN reacted.
Chemical equilibrium is a point in a chemical reaction when substances form and break down at
the same rate, and the number of molecules of each substance become constant. An Equilibrium
constant value (Kc) is calculated with the molar concentrations of the products and the reactants at
chemical equilibrium. A large equilibrium constant implies that the chemical system occurring is
products favored. The equilibrium constant value (Kc) is determined by dividing the equilibrium
concentrations of the products by the concentration of the reactants. (Equation 3)
Knowing if a chemical system is products favored is crucial to scientists. If a product of a
certain chemical reaction is needed, it is important to find out the best way to get this product. If the
equilibrium constant is less than one, the chemical system is reactants favored and is not profitable or
beneficial to continue it. If its larger than one, it is a products favored reation.
In the calibration experiments of part A, the goal is to correlate the amount of thiocyanatoiron
(II) ion complex produced with the absorbance values determined by spectrophotometric analysis of
these standardized solutions. To ensure that the amount of FeNCS2+ present in solutions is equal to the
initial amount of SCN- placed in the solution, the molar concentration of Fe3+ must be in excess to push

the equilibrium reaction far to the products side of the reaction. (1). This implies that the initial amount
of moles of SCN in a solution will result in an equal amount of moles of FeNCS2+ present when combined
with Fe3+ solution.
The spectrophotometric absorbance values will be correlated with each respective sample of
varying FeNCS2+ molar concentrations and plotted to establish a Beer lambert calibration curve. This
curve gives a relation between how much FeNCS2+ is produced with how much light is absorbed by the
sample. The molar concentration of FeNCS2+ (at equilibrium) in the samples of Part B can then be
determined by spectrophotometric analysis.
Experimental Procedure Part A
Preparation of Standardized Solutions
10 mL of 0.2 M Iron (III) ion solution was measured out in a 10 mL graduated cylinder. Six (6)
separate, marked and clean 25 mL volumetric flasks were filled with 10 mL of Iron (III) solution. . Each
respective flask was placed in order and 10 mL graduated cylinder was washed and dried. 1.0 mL of
thiocyanate ion solution is placed in sample 1, 2.0 mL in #2, 3.0 mL in #3, 4.0 mL in #4, and 5.0 mL of
Thiocyanate Ion solution in #5. One 25 mL flask, (marked blank) did not receive any thiocyanate ion
solution. Remaining volume of each prepared 25 mL volumetric flask was filled to 25 mL mark with 0.1
M nitric acid. Clean stoppers were placed on all (6) 25 mL volumetric flasks and flasks were inverted and
agitated for approximately 30 seconds to ensure uniformity. Initial Molar Concentrations of each
respective solution and sample were calculated and recorded on data sheet.

Spectrophotometric Analysis (Part A)

Six (6) cuvettes were washed with water and dried prior to addition of standardized solutions.
Cuvettes were inspected for scratches and discoloration. Defective cuvettes were discarded. Each
prepared solution were placed in separate, unblemished, cuvettes and filled to approximately of the
cuvettes volume. Cuvettes were capped and placed on a sheet of paper marked with sample numbers.
Cuvettes were transferred to spectrophotometer. Cuvette containing Blank solution was wiped with Kim
wipes and aligned with clear side facing light source. Spectrophotometer was calibrated to desired
wave length of 447 nm. Blank was removed. Each of the remaining five standardized solutions were
wiped with Kimwipes prior to placement into spectrophotometer and each respective absorbency value
was recorded on data sheet prior to moving on to subsequent cuvettes. Cuvettes were saved until
calibration curve was established.
Establishing Beer Lambert Calibration Curve
Absorbance values recorded on data sheet were inputted in to Excel spreadsheet to determine
coefficient of determination (R2) and Beer Lambert Law equation. Absorbency values were placed
with correct initial molar concentrations of each standard sample. Coefficient of determination must be
in an acceptable range for standard solutions to be valid.
Experimental Procedure Part B
Preparation of Test Solutions
5 mL of 0.002 M Iron (III) ion solution was measured out in a 10 mL graduated cylinder. Six (6) separate,
marked (6, 7, 8, 9, 10 and Blank) and clean 10 mL volumetric flasks were filled with 5 mL of Iron (III)
solution. Each respective flask was placed in order and 10 mL graduated cylinder was washed and dried.
1.0 mL of .002 M Thiocyanate Ion solution was placed in flask marked # 6, 2.0 mL of .002 M Thiocyanate

Ion solution was placed in flask marked #7, 3.0 mL of .002 M Thiocyanate Ion solution was placed in flask
marked #8, 4.0 mL of .002 M Thiocyanate Ion solution was placed in flask marked #9, 5.0 mL of .002 M
Thiocyanate Ion solution was placed in flask marked #10. One 10 mL flask, (marked blank) did not
receive any thiocyanate ion solution. 10 mL graduated cylinder was washed and dried, in preparation of
Nitric Acid measurements. 4.0 mL of 0.1 M nitric acid was added to flask #6, 3.0 mL added to #7, 2.0 mL
added to #8, and 1.0 mL was added to #9. Clean stoppers were placed on all six (6) 10 mL volumetric
flasks and were inverted and agitated for approximately 30 seconds to ensure uniformity.
Spectrophotometric Analysis (Part B)
Six (6) cuvettes were washed with water and dried prior to addition of standardized solutions. Cuvettes
were inspected for scratches and discoloration. Defective cuvettes were discarded. Each prepared
solution were placed in separate, unblemished, cuvettes and filled to approximately of the cuvettes
volume. Cuvettes were capped and placed on a sheet of paper marked with sample numbers. Cuvettes
were transferred to spectrophotometer. Cuvette containing Blank solution was wiped with Kim wipes
and aligned with clear side facing light source. Spectrophotometer was calibrated to desired wave
length of 447 nm . Blank was removed. Each of the remaining five standardized solutions were wiped
with Kimwipes prior to placement into spectrophotometer and each respective absorbency value was
recorded on data sheet prior to moving on to subsequent cuvettes. Cuvettes were transferred back to
lab table and saved until data was established.

Calculations
Part A. (Standard Solution #1) See Table 1 for calculations and data for samples 2 - 5
SCN- (mols)
.000001
[SCN-]
.000025 M
[FeNCS2+]
.000025 M

=
=
=

Vol. NaSCN
.001L
SCN- (mol)
.000001
[SCN-]
.000025 M

.001 M NaSCN
.001
.025 L
.025 L

Part B. (Test Sample # 6) See Table 2 for calculations of samples 7 - 10

Fe3+ (mols) initial
.00001
SCN (mols) initial
.000002

=
=

Vol. Fe(NO3)3
.005 L
Vol. NaSCN
.001 L

.002 M Fe(NO3)3
.002
.002 M NaSCN
.002

Part C. (Kc value for Test Sample # 6) See Table 2 for calculations of 7 - 10
[FeNCS2+ ](Eq)
.000041
3+
Fe (mols) (reacted)
.00000041
SCN (mols) (reacted)
.00000041
3+
Fe (mols) (Eq)
.00000958
SCN- (mols) (Eq)
.00000158
3+
[Fe ] (Eq)
.000958
[SCN-] (Eq)
.000158

Kc =

= Abs. + 0.0154
.098 + 0.0154
= [FeNCS2+ ](Eq)
.000041
= [FeNCS2+ ](Eq)
.000041
3+
= Fe (mols) initial
.00001
= SCN (mols) initial
.000002
3+
= Fe (mols) (Eq)
.00000958
= SCN- (mols) (Eq)
.00000158

[FeSCN 2+ ]
[Fe3+ ][SCN ]

269.08

2764.3
2764.3
0.010 L
0.010
0.010 L
0.010
Fe3+(mols) (reacted)
.00000041
SCN- (mols) (reacted)
.00000041
0.010 L
0.010
0.010 L
0.010

(3)

Experimental Data
Table 1. Data and Results from Thiocyanatoiron Ion Complex Calibration Solutions
Solution #

Blank

Vol. NaSCN (L)

SCN (mols)initial
[SCN-]initial
[FeNCS2+]Eq
Absorbance

0
0
0
0
0

1
.001
1.0 x 10-6
4.0 x 10-5
4.0 x 10-5
0.076

2
.002
2.0 x 10-6
8.0 x 10-5
8.0 x 10-5
0.214

.003
3.0 x 10-6
1.2 x 10-4
1.2 x 10-4
0.306

.004
4.0 x 10-6
1.6 x 10-4
1.6 x 10-4
0.422

.005
5.0 x 10-6
2.0 x 10-4
2.0 x 10-4
0.548

Table 2. Data and Results from Thiocyanatoiron Ion Complex Test Solutions
test sample

[FeNCS2+]Eq
4.1023 x 10-5
3+
[Fe ]Eq
0.000958977
[SCN ]Eq
0.000158977
Absorbance
0.098
Kc
269.08
Average Kc = 309.30

9.0945 x 10-5
0.000909055
0.000309055
0.236
323.71

0.0001311
0.000168723
0.0008689
0.000831277
0.0004689
0.000631277
0.347
0.451
321.78
321.52
Std. Deviation () = 23.08

10
0.00019911
0.00080089
0.00080089
0.535
310.42

0.6

Absorbency Value (Abs. )

0.548
0.5
0.422

0.4
0.306

0.3
0.214

0.2
0.1
0

0.076
0

0.00005

0.0001

0.00015

Molar Concentration [FeNCS

0.0002
2+]

Figure 1. Beer- Lambert Calibration Curve (absorbance values of 447 nm wavelength)

2
Coefficient of Determination (R ) = 0.9957

0.00025

Results and Discussion

Spectrophotometric Analysis of Thiocyanatoiron (III) Standardized Solutions
Using the methods previously described in experimental procedure Part A, the anticipated
equilibrium molar concentrations of Thiocyanatoiron (III) Ion complex in the standard samples (1
through 5) are listed in Table 1. The spectrophotometer emitted the specified EMR wavelength of 447
nm and spectrophotometric data of each respective sample is also listed in Table 1. Absorbance values
for the Thiocyanatoiron (III) Ion complex standard solutions were plotted versus the expected
equilibrium concentration values of FeNCS2+ to construct the Beer Lambert Calibration Curve (Figure
1). First attempt at obtaining absorbency values to determine of Beer Lambert Calibration curve
resulted in a satisfactory coefficient of determination (R2 value) of .9957. Subsequent standardization
was not performed. Beer Lambert Calibration Curve resulted in a slope formula of Absorbance Value
(Abs.) equation 4:

Abs = 2764.3([FeNCS2+] 0.0154

(4)

Spectrophotometric Analysis of Equilibrium Test Solutions

In part B of the experimental procedure, five samples with various initial concentrations of Iron
(III) ion solution and Thiocyanate Ion solution were prepared. Each test sample (6 through 10) was
prepared in accordance with procedures listed in Part B of the experiment. Absorbance values 0.076,
0.214, 0.306, 0.422, and 0.548 were obtained from spectrophotometer set to deliver 447 nm
wavelengths. These values were recorded after stabilization of digital read out was achieved. The Beer
Lambert calibration curve, derived in part A (Figure 1), was used to determine each respective test
samples thiocyanatoiron (III) ion equilibrium concentration. Calculations for estimated equilibrium
concentrations by spectrometric analysis for samples 6 through 10 were determined in the same

manner as calculations listed in corresponding calculations section. Test sample #6 deviated drastically
from the four other estimated equilibrium constant (Kc) values. The remaining findings are reported in
Table 2.
The average equilibrium constant (Kc) determined by spectrophotometric analysis of test
samples resulted in a value of 309.30. (Table 2.) Expected equilibrium constant of 271.49 and observed
value were used to determine the percent error. 13.9% error was obtained by using equation 5 below.
% . =

(5)

100

Possible explanations for margin of error may have resulted from methods of measurement,
temperature changes of solutions and possible human error.
The dispersion of the equilibrium constant values determined in Part B were unsatisfactory. The
standard deviation was determined by taking the difference between the average Kc value and the

(6)

particular estimated Kc value [represented as (d)]. N is the number of test solutions in the experiment.
Equation 6 was used to determine a standard deviation () of 23.08. This value expresses the amount
by which any future readings will deviate from subsequent values.
The use of a graduated cylinder may have led to imprecise measurements of the initial volume
of reactants. This error in volume may have initiated the elevated percent error from the expected
equilibrium constant for Thiocyanatoiron Ion solution. Using a Mohr pipet or buret may have led to a
more precise molar concentration of Thiocyanatoiron Ion complex present in standard and test samples
Also, the preparation of the standardized and test solutions may have been affected by human error.

Inexperience of lab team members/students, and the number of lab members taking various
measurements, may have led to inconsistent initial concentrations of the reagents in this experiment.
From further evaluation of the Beer Lambert calibration curve an adjustment by omitting
standard sample 1 (the one with the greatest deviation from trend line) increased the R2 value from
.9957 to .9977. This standard solution had the smallest amount of NaSCN ion solution and may have
been affected the most by imprecise measurements. The resulting slope line equation was Abs =
2706.3([FeNCS2+] 0.0051. This more precise R2 value and slope equation lowered the average
equilibrium constant (Kc) from 309.3 to 302.02 but increased the standard deviation of the test solutions
from 23.08 to 32.05. The resulting percent error dropped by 2.7 percentage points to 11.2%

Conclusion
The purpose of the experiment is to be able to determine an equilibrium constant for the
chemical reaction between iron ion and sodium thiocyanate solutions from spectrophotometric analysis.
The calibration curve derived from part A shows that the correlation between absorbance and
concentration does exist. Incremental increases in thiocyanatoiron concentration did result in higher
absorbance values. The Beer Lambert calibration curve, the average Kc value of 309.3, and increase in
color intensity of the solution provides evidence that the resulting thiocyanatoiron solution was a
products favored reaction

Works Citied
(1)

Beran , Jo Allan. (2011). Laboratory Manual for Principles of General Chemistry. Hoboken NJ.
John Wiley & Sons .

(2)

Tro, Nivaldo J. (2011). Chemistry - a Molecular Approach. Upper Saddle River, NJ. Pearson
Prentice Hall.

(3)

Brooker, Robert J. (2011). Biology 2nd ed. New York, NY. McGraw Hill.

(4)

Beers Law and Spectrophotometric Analysis. General Format. Retrieved from

www.chem.ucla.edu/~gchemlab/colormeteric

(5)

Spectrophotometer. General Format. Retrieved form

www.ncbionetwork.org/spectrophotometer