CHE 314

TEFO OLEFILE
201502191
EXPERIMENT 3
SPECTROPHOTOMETRIC DETERIMINATION OF
IRON IN VITAMIN TABLETS
WEEK 3
DATE OF SUBMISSION: 13 MARCH 2018
GROUP: TUESDAY 3-6PM
SPECTROPHOTOMETRIC DETERMINATION OF IRON IN VITAMIN
TABLET

AIM
The aim of this experiment is to use the quantitative technique of spectrophotometry
to determine the mass of iron contained in a commercially available vitamin tablet as
well as its molar absorptivity.

ABSTRACT
The aim of this experiment is to use the quantitative technique of spectrophotometry
to determine the mass of iron contained in a commercially available vitamin tablet as
well as its molar absorptivity. The unknown solution was prepared by adding an iron
containing tablet and HCl in a beaker and boiled gently and later on filtered.Standards
at different concentrations were prepared in 100ml flasks. A calibration curve was
prepared using a spectrophotometer and the molar absorptivity of the iron complex
was determined.The concentration of the unknown was found to be 2.04x10 -6 mol/L
using Beer-Lambert law. The molar absorbtivity was found to be 1.08 x 10 4 cm-1mol-
1
L using the concentration of the 2ppm standard of the iron solution and the cell path
length. The amount of milligrams of iron in unknown was 1.1393 mg by Beer`s law
and approximately 1.5mg from the graph interpolation

INTRODUCTION
Iron is element is very useful in biological systems .In the human body it is mostly
found in the blood in a protein called hemoglobin used for oxygen and carbon dioxide
transportation during respiration. The iron necessary for the formation of hemoglobin
is obtained from the diet in foods such as meat and leafy, green vegetables. When the
dietary intake is deficient in iron, a condition called anemia results. Anemic people
exhibits lack of energy and often has an unusually pale skin tone (the red color of
blood is also a result of the presence of iron in hemoglobin). Dietary supplements of
iron in the form of vitamin tablets can be administered to help alleviate this condition.
(McMaster,2007)

In this experiment iron content in the vitamin tablets is analysed using the technique
of spectrophotometry. It is one of the most common techniques used in the
quantitative analysis of samples for a specific chemical substance,which uses an
instrument called a spectrometer.Spectrophotometric methods exist for the
quantitative determination of practically every element on the periodic chart as well as
a large number of compounds. Many standard methods of analysis approved by the
FDA, EPA, and clinical diagnostic work depend on the selective absorption of visible
radiation by any ionic or molecular species which exhibits a characteristic color in
solution. This analytical technique dates back to 1838 when it was called
“colorimetry-or measuring color.” In most methods of absorption spectrophotometry
the absorption spectrum provides a “finger print” of the chemical species for
qualitative purposes. Beer’s Law developed in 1852 establishes the quantitative
relationship between the amount of radiation absorbed and the concentration of the
absorbing chemical species. Beer’s law may be expressed as
A = εcl
where A is absorbance,  is the molar absorptivity (M-1cm-1), b is pathlength (cm), and
c is molar concentration.
Iron from a vitamin supplement tablet is dissolved in hydrochloric acid and then
reduced to Fe 2+ with hydroquinone:

2 Fe 3+ + HO OH 2 Fe2+ +O O + 2 H+

While freshly-dissolved Fe2+ is nearly colorless an intense red color can be

imparted by a stoichiometric reaction of Fe2+ with three molecules of o-
phenanthroline (phen):

Because the molar absorptivity value is not always a known quantity, a calibration
curve is often constructed from a series of standard solutions. A standard solution is
one in which the concentration of the species being analyzed is known. The
absorbances of several standard solutions are measured, and these values are plotted
as a function of the concentrations of the solutions. If the absorbing species behaves
according to Beer’s law, such a plot should produce a straight line. The absorbance of
a solution of unknown concentration can then be measured with the
spectrophotometer, and this value can be used in conjunction with the calibration
curve to determine the concentration of this solution.(Christian,2004)

REAGENTS AND APPARATUS

REAGENTS AND APPARATUS

Table 3.0: Reagents and apparatus used

Reagents Apparatus

1 iron containing vitamin tablet Analytical balance

0.0300 g of ferrous ammonium sulphate Weighing boat
Distilled water 7 x 100 mL volumetric flask
Concentrated sulphuric acid 1ml, 5ml, 10ml and 25 ml pipettes
1, 10-phenanthroline solution Filter paper
Hydroxylammonium chloride solution Hot plate
Sodium acetate solution 100ml beaker
6M hydrochloric acid UV-Vis spectrophotometer
Unknown sample

PROCEDURE
a.Preparation of iron (ii) standard stock solution

0.0281 g of ferrous ammonium sulphate was measured and placed into a 100 mL
volumetric flask. Water was added to dissolve the salt. 1.0ml of concentrated
sulphuric acid was also added and more distilled water to dilute to the mark. This
solution contained 40 mg per litre (40ppm).

b. Sample preparation

A vitamin tablet containing iron was placed in a 100ml beaker with 25ml of 6M
HCL.This was gently boiled on a hotplate in the fume hood for about 15 minutes .The
solution was filtered directly into the 100ml volumetric flask using gravity filtration.
It was allowed to cool then diluted to the mark using distilled water. 10.0 ml of the
sample was then pipetted into another 100ml volumetric flask.The following volumes
of standard iron solutions were also pipetted into labelled volumetric flasks: 1.00,
2.00, 5.00, 10.00ml. In the blank flask 50.0mL of distilled water was added to serve as
a blank. 1.0 mL of the hydroxyl ammonium chloride solution and 5.0 mL of the 1.10-
phenanthroline solution were added to each of the flasks including the unknown. Each
solution was buffered by addition of 8.0 mL of sodium acetate solution to produce the
red colour of ferrous 1.10-phenanthroline. These solutions were left for about 15
minutes for the complex to form. Solutions were then diluted to the mark with
distilled water. The absorption spectrum of the 2ppm (5 mL of stock in 100 mL)
solution was used to obtain the absorbance from 400nm to 700nm.

RESULTS AND ANALYSIS

Table 3.1: Absorbance For Standards At 509.3 and The Calculated Concentrations

Standards(mL) Absorbance Concentration (mol/L)

1.0 0.076 7.41 x 10-6

2.0 0.158 1.45 x 10-5

5.0 0.419 3.61x 10-5

10.0 0.829 5.88 x 10-5

Unknown 0.034 2.04 x 10-6

Sample calculations
 a. Concentration of Fe in Iron standard solution

Mass = 0.0281g,molar mass= 392.70248g/mol

mass
n=
mm

moles = (0.0281 g) / (392.70248 g/mol) = 7.156 x10-5 mol

Therefore,

n
conc=
V

= (7.156x10-5 mol) / 100 x 10-3 L = 7.1554x10-4 mol/L = 7.16 x 10-4 mol/L

b. Concentration of the 2ppm standard solution using dilution law

M1V1 = M2V2

[(7.16 x10-4 mol/L) x 5 x 10-3 L)]= M2 x (100 x 10-3 L)

M2 = 3.58 x10-5 mol/L

Now, molar absorbtivity can be calculated from Beer-Lambert’s law

A=εbc

Where A, is absorbance at maximum wavelength (510.8 nm), i.e. 0.390

ɛ = A/bc = 0.419 / (1cm x 3.58 x10-5 mol/L) = 11703.9 cm-1mol-1L

= 1.17 x 104 cm-1mol-1L

c.Concentration of standard 1 using Beer lambert`s law

C=A/b ɛ

=0.076/(1x1.08x 104)

=7.41x10-6

NB; all the other concentrations were calculated using the method above

2.Concentration from the graph

Y=0.039+(0.154)C

Y=0.390

Therefore 0.390=0.039 +0.154C

C=(0.390-0.039)/0.154

=2.2792 mg/L

Calculation of the mass of Fe in solution

Mol = concentration x volume

=3.58 x10-5 mol/L x 0.1L

=3.58 x10-6 mol

Mass=mol X mm

=3.58 x10-6 mol X 55.845g/mol

=2.0 x 10-4 g

=0.2mg
Concentration of unknown using Beer-Lambert’s law.

C = A/ɛb = 0.022/ (1x 1.08x 104) = 2.04 x 10-6 mol/L

M1 V1 = M2 V2

M1 = [(2.04 x10-6 mol / L) x (100 x 10-3L)] / 10 x 10-3L

M1 = 2.04 x 10-4 mol/L

N=CxV

= (2.04 x 10 -4 mol/L) x (100 x 10-3 L)

= 2.04 x 10 -5 moles

N = mass/mm

molar mass of iron = 55.847 g/mol

Mass = n x mm

= 2.04 x10 -4moles x 55.847 g/mol

= 0.001139278 g

Hence, concentration = 1.1393 mg/L.

 Graph 1: absorbance for iron ii against concentration

0.7
f(x) = 0.158 x − 0.2216
R² = 0.932030889483926

0.6

0.5

0.4
absorbance

0.3

0.2

0.1

0
0 1 2 3 4 5 6 7

concentration in mg/L

The concentration from the graph is ~ 1.5mg/L .

DISCUSSION
The 2ppm standard iron solution was used to determine the maximum absorbance or
wavelength. These were found to be 510.8nm and 0.390 respectively. All the other
standards and unknown were measured using the same wavelength.From the results
obtained,as concentration increased so did the absorbance showing that more and
more of light was trapped as the iron complex particles increased. At 25ppm of the
standard though it was expected an increase in absorbance and there was a sharp
decrease in the absorbance value due to the lack of consistency in concentration
increase. This value affected the linearity of the graph thus it was removed and value
of the R2 was equal to 0.986. The molar absorbtivity was determined as 1.08 x 104 cm-
1
mol-1L using the concentration of the 2ppm standard of the iron solution and the cell
path length. This molar absorptivity was 1.81% less than the literature value 1.10 x
104 cm-1mol-1L.This value was almost accurate hence it could be further used in
calculations. The calculated molar absorptivity was used to calculate the concentration
of the standard solutions and there was direct proportionality between concentration
and absorbance,that is as concentration increased absorbance increased also increased.
This agrees with Beer`s Law and with literature that absorbance is directly
proportional to concentration.(Daniel,2008).When using the graph the amount of iron
in the tablet was found to be aprroximately equal to 1.5 mg/L while calculation by
Beer`s law gave a concentration of 1.1393 mg/L.The two concentrations differed by
almost 24% and this could have been due to errors in the experiment.Possible sources
of error include errors in calculations or measurements during process of pippeting as
well parallex error during dilution resulting in less or higher concetration than it
should be.Also the uncertainty of the instruments has to be taken into account. The
stray of light, noise and effects due to polychromatic radiation may cause
spectrophotometers to suffer from non linearity(Skoog,1998). The selection of λmax
may also be a contributing factor in this experiment. The λmax used in this experiment
was 510.8 which differs from the 508nm from literature.This could be the reason why
there was a difference in the absorptivity since the one in literature was determined at
508nm. Proper and accurate measurements and good care handling of the solutions
and reagents can help in reducing these errors. In addition, it is important that the
acidity of the solution is carefully controlled the presence of sodium citrate in solution
will neutralize some of the acid and maintain the proper pH. Other sources of error
fingerprints on the cuvette which reduces the transparency of the cuvette.
CONCLUSION
The concentration of the unknown was found to be 2.04x10-6 mol/L using Beer-
Lambert law. The molar absorbtivity was found to be 1.08 x 104 cm-1mol-1L using the
concentration of the 2ppm standard of the iron solution and the cell path length. The
amount of milligrams of iron in unknown was 1.1393 mg by Beer`s law and
approximately 1.5mg from the graph interpolation.

REFERENCES
1. Christian, G.D., Analytical Chemistry, 6th edition. New Jersey. John Wiley,
2004
2. Daniel C. Harris’ Quantitative Chemical Analysis and R. C. Atkins,2008
3. McMaster, Marvin C. Apractical User's Guide. 2nd. New York: John Wiley &
Sons, Inc., p130,2007
4. Skoog, D. A.; Holler, F. J.;Nieman, T. A. “Principles of Instrumental Analysis’’, 5th
ed.; Harcourt Brace & Company: Philadelphia, 1998