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EXPERIMENT 1
Spectrophotometric Determination of Fe2+ Ions using 1,
10-Phenanthroline
(External Calibration Method)
Vanessa Olga J. Dagondon and Ken M. Menez
Department of Chemistry, College of Arts and Sciences,
University of the Philippines Visayas, Miag-ao Iloilo
ABSTRACT
The determination of the Fe 2+ concentration in a branded iron
supplement tablet was developed using the external calibration
method. This method uses a series of standardized solutions to
obtain data through the UV VIS spectrophotometer to construct a
graphs that relate the wavelength to absorbance and, ultimately, to
the concentration of a solution. The standard used was a hydrated
derivative of ferrous sulphate, which is called ferrous ammonium
sulphate hexahydrate. Because of ferrous ions typically exist as
having a sulphate base in compounds; it was assumed that the iron
tablet contained mainly a high percentage of this ferrous sulphate.
This experiment also tackled to explain the principle as to how a
solution absorbs light, and how this could be related to the
concentration of ions in the solution. This experiment had also
provided first-hand experience to operating a UV VIS
spectrophotometer and practice to calculating necessary data
through the least squares method. This least squares method was
considered important at the event that an automatic calculating
device was not on hand. The principles followed by each property of
absorbance follow the Beers Law, which is reintroduced and
expanded in definition and use in this experiment. Lastly, since this
was a class experiment, the Fe 2+ concentration obtained by each
group was correlated and compared with each other in the whole
class. This was performed to verify the reliability of the method for
determining the said Fe2+ concentration.
INTRODUCTION
Spectrophotometric methods are based on the interaction of a
sample and of the light passing through it. These methods measure the
amount of light absorbed by the sample and thereby identifying the
amount or concentration of a part of the sample or of the sample which
interacts with it.
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Then, the weight of iron present in the tablet was calculated using the
concentration of the sample multiplied by a number of dilution factors.
RESULTS
AND
DISCUSSION
Absorbane
0.2
0.15
0.1
0.05
0
400
-0.05
450
500
550
Wavelength (nm)
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600
650
0.2
0.15
0.1
0.05
0
0
0.5
1.5
Concentration (ppm)
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2.5
for the cuvette used has a constant value of 1.00 centimeter, which
effectively cancels its value from the Beers Law equation.
Fig. 2 shows the plot of the absorbance against the concentration of
the prepared standard ferrous solutions. This calibration curve describes
the linear relationship between the absorbance and concentration. The
slope of the line is the proportionality constant between the absorbance
and concentration of the standard solutions. The resulting slope is equal
to the molar absorptivity calculated using Beers Law. This also denotes
how the absorbance of a solution is directly proportional to its
concentration as a suggested in Equation 1. It was observed that as the
concentration of the analyte increases in each of the standard solution,
the color intensity also increases. This indicated a direct proportionality
between the two components.
The standardization technique used in the experiment was a multipoint external standard calibration method. It is widely used to analyze a
series of samples using a single calibration curve. In this technique the
samples matrix and that of the standard solutions matrix is assumed to
be the same and that would not affect the value of the proportionality
constant. However, this is not always the case and proportional
determinate error is introduced into the analysis. This is a limitation in this
kind of technique.
The equation of the line, obtained by least squares method, in the
calibration curve contained the value for the concentration of Fe 2+ in the
sample solution. This equation is in the form y = mx + b, where y
corresponds to the absorbance of the sample solution, m is the slope or
proportionality constant, b is the y intercept gained from the linear
regression process, and x is the concentration of the sample. The
concentration of the sample solution was then multiplied to a series of
dilution factors to obtain 61.7624 mg Fe2+.
The obtained concentration of ferrous ion in the sample is analyzed
statistically. Originally, triplicates of the sample should have been
analyzed however due to time and budget constraints only a single
sample was analyzed. The standard deviation from the results of the
calibration curve (sc) was calculated to measure the reliability of the result
of the external standard calibration. The standard deviation in the
regression line (sr), the relative standard deviation (RSD) was also
calculated. To measure the linearity of the calibration curve, R 2 was
calculated. Table 1 summarizes the statistics done in the groups sample.
The class standard deviation of the obtained concentration of the
ferrous ion in the sample was calculated. Also, the relative standard
deviation and coefficient of variance were determined to measure the
precision of measurements. Using Grubbs test on the concentrations of
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the five class sample solutions, there were no outliers detected in the
class data. Table 2 summarizes the statistics done on the class data.
CONCLUSION
The method used to standardize the sample was the external
standard calibration method.
The concentration of Fe2+ ion in the sample of 325 milligrams of
ferrous sulfate was calculated to be 61.7624 mg/tablet. The s c was equal
to 0.02235, representing reliability for the external standard calibration
method to produce data points with precision. The mean class data for the
concentration of the sample solution had an average of 61.7722
mg/tablet. The standard deviation for the whole class was calculated to be
1.9666. This suggests that there is a slight variation in the results relative
to the mean of the class data.
LITERATURE CITED
Harvey, D. May 2002. External Standards or Standard Additions? Selecting
and Validating a Method of Standardization. Journal of Chemical
Education Vol. 79: 613-615.
Harvey, D. Modern Analytical Chemistry. United State of America: The
McGraw-Hill Companies, Inc.; 2000 [cited 2016 February]. Available
from: http://elibrary.bsu.az/
APPENDICES
x
5.6616
Sxx
x
7.8090
Syy
TABLES
y
1.1590
Sxy
3.2299
sr
0.1267
Xunknown
0.6394
sc
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y2
0.3186
Slope (m)
0.1980
RSD
xy
1.5769
Y
intercept
(b)
0.005448
R2
0.004041
61.7624
0.02235
0.0003618
Table 1. External Standard Calibration Method Group Data
xsample
ssample
sample
0.9994
RSDsample
308.8608
61.7722
1.9666
0.0318
Gmax
Gmin
Gexp
CV
1.0147
1.4148
1.5712
3.1836
Table 2. External Standard Calibration Method Class Data
CALCULATIONS
x= x i=0.1011+0.2022+0.3033+ 0.5055+ 1.011+1.5165+2.0220=5.6616
x 2= xi2 =0.0102+ 0.0408+0.0920+0.2555+1.0221+2.2998+ 4.0885=7.8090
y=y i=0.029+0.042+0.068+ 0.103+0.202+0.311+0.404=1.1590
2
( x )2
( 5.6616 )2
=7.8090
=3.2299
n
7
( y )2
( 1.1590 )2
S yy = y
=0.318579
=0.1267
n
7
2
( xy )2
[(5.6616)(1.1590)]
S xy =xy
=1.5769
=0.6394
n
7
m=
S xy 0.6394
=
=0.1980
S xx 3.2299
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0.204222 +
b=
y
x 1.1590
5.6616
m
=
( 0.1980 )
=0.005448
n
n
7
7
( )
y b
m
sr =
0.250.005448
=61.7624
0.1980
x unknown=
2
S yy m2 S xx
0.1267( 0.1980 ) (3.2299 )
=
=0.004041
n2
72
( m
S xx |=
RSD=
x unknown
R =1
0.25
1.1590
7
0.004041 1 1
+ +
=0.02235
0.1980 1 7 ( 0.19802 ) ( 3.2299 )
1 1
+ +
M N
sr
m
s c =
sc
y
n
0.02235
=0.0003618
61.7624
[ y i( b +m x i ) ]
S yy
=0.9994
x sample =
x sample 308.8608
=
=61.7722
n
5
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( x ix )
( 58.989961.7722 )2+ ( 63.767761.7722 )2 + ( 63.535461.7722 )2+ ( 6161.7722 )2+ ( 6
s sample =
=
n
5
1.9666
RSD sample =
ssample 1.9666
=
=0.0318
x sample 61.7722
Gmax =
Gmin =
Gexp=
( )(
n1
n
) ( )(
CV 2
51
=
2
n2+CV
5
3.1802
=1.5712
52+3.1802
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