NAME: NOOR AWAN

SUBMITTED TO: MAM ANUM KHALEEQ

SUBJECT: ANALYTICAL CHEMISTRY

DATE OF SUBMISSION: 21ST FEBRUARY, 2023

TOPIC: DETERMINATION OF Fe3+ BY MEANS OF

CALIBRATION DIAGRAM
1 Outlining 1
2 Sequence 1
3 Organization 1
4 Results and conclusion 1
5 No. of pages 0.5
6 Turnitin Report 0/5
Total 5

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TABLE OF CONTENTS:

SR#1 INTRODUCTION

SR#2 THEORY

SR#3 PROCEDURE

SR#4 APPARTUS

SR#5 REAGENT

SR#6 PREPARATION OF
STADARD SOLUTION
SR#7 REACTIONS

SR#8 OBSERVATION

SR#9 CALCULATIONS

SR#10 CALIBRATION CURVE

SR#11 RESULTS

SR#12 CONCLUSION&
REFERENCES

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 DETERMINATION OF AMOUNT OF Fe3+ BY MEANS OF
CALIBRATION DIAGRAM
INTRODUCTION:
Iron is essential and very important compound in our body. Hemoglobin, the protein responsible
for the transport of oxygen throughout our body, contains iron.8mg iron per day is the daily
requirement of male aged between 18 to 30 years of age. While in case of female, the
requirement is 18 mg per day. Deficiency or iron causes anemia and other diseases .It can be
determined with help of a number of methods. These method is used to know the amount of iron
in blood and in samples.

THEORY:
The percentage of iron in a sample is calculated with using a spectrometer instrument, beer law
and by using a set of standard solution. A stable red complex is formed by the reaction of iron
with three o- phenanthroline bidentate ligand. This is done by converting to another form of iron
with the usage of 10 percent hydroxyl amine hydrochloric acid. The standard solution is formed
by the iron complex.

BEER LAMBERT LAW:

This law states that there exists a straight relationship between the concentration and the
absorbance and optical coefficient of a solution and molar coefficient of absorption. The
mathematical notation is as follows:

A=εbc
Where a designates absorbance, b gives a symbol for solution path length, c for concentration
and ε stands for the coefficient of extinction.

CALIBRATION CURVE:
This is basically used for the concentration determination for knowing the substance that is not
known. It includes plotting of points at different concentrations. Various researchers and
scientists employ this method.

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SPECTROPHOTOMETER:
An instrument called the spectrophotometer can be used to determine the amount or
concentration of an already knowing substance by means of the intensity of the detected light.by
using the photons number, the intensity of light can be measured and it is absorbed. By using this
information, the amount is determined. By passing the light bean through the sample, intensity of
light is measured.

SAMPLE MONOCHROMATOR SAMPLE

READOUT( METER OR
DETECTOR
RECODER)

Fig 1: Block Diagram of a Spectrophotometer

PROCEDURE
0.2500 g\L of the pure iron was present in the standard solution of iron. In a volumetric flask, to
make a solution of 500 mL the pipet out was 25.00 mL, the distilled water was diluted up to the
mark. The iron calibrated amount were prepared by taking out a designed indicated amount in a
50 ml measuring flask. The one that contains iron is first one and it is named as blank. This all is
listed in the observation table. Into a 250ml volumetric flask,10ml of unknown solution was
pipette out and it was filled with distilled water up to the mark.in order to mix the solution the
flask was mixed several times. The unknown solution was taken into two halves after splitting
the two 50 mL flask that was labelled.To each of the volumetric flask with help of a 10 mL
graduated cylinder, add in it hydroxylamine hydrochloride 4 ml, add in it phenonthroline
solution as well. Let the mixtures for rest and stand it for about 10 minutes. Each flask was
diluted and distilled water was added up to the mark and it was mixed and shaked well
thoroughly. With help of a spectrophotometer, the percentage of transmittance of various
solutions was calculated. The result was recorded in the results table. A graph was plotted for the

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absorbance versus the concentration of the original solution. The graph was plotted and drawn
with the best fitting line. This plotting on the graph is termed as Beer Lambert Law. The
concentration was determined by plotting the absorbance value of the unknown solution. The
amount of iron in the unknown sample was calculated.

APPARATUS:
 A standard iron solution prepared in a 500cm 3 volumetric flask having a known amount
of iron
 Unknown amount of iron in a 250cm3 volumetric flask
 Smaller flasks containing different standards of iron added in a flask
 A double beam vision spectrophotometer
 A microcomputer for comparing the results

REAGENTS:
 Ferrous Ammonium Sulphate Hexahydrate
 (10 g in 100 ml of distilled water) Sodium Acetate
 (10 g in 100 ml of distilled water) Hydroxylamine hydrochloride
 (0.1 of 1.10 phenanthroline in 100 ml of distilled water) o- phenanthroline
 Warming water if necessary to effect the solution

PREPARATION OF STANDARD SOLUTION:

For the preparation of standard solution, ferrous ammonium sulfate was weighed out having an
accuracy of 0.1 mg. 0.00200M preparation of solution for 250 ml with respect to that compound.
For the preparation of stock iron solution, 3M H 2SO4 was added to 8ml. the salt was transferred
to a volumetric flask. The 10ml of stock iron solution was pippete out to a 100 ml volumetric
flask, it was mixed well by adding the 3ml of 3M H 2SO4 and distilled water was added up to the
mark. This was named to be as original iron solution and the amount of iron was calculated in
it.All the solutions with analytical purity were prepared.

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REACTIONS:

OBSERVATION:
Concentratio Volume to Solution Transmittances Absorbance

n of Iron pipet 0.00 mg iron 100% 0%

0.00 mg iron 0.00mL 0.05 mg iron X X

0.05 mg iron 4.00mL 0.10 mg iron X X

0.10 mg iron 8.00mL 0.15 mg iron X X

0.20 mg iron X X
0.15 mg iron 12.00mL
0.25 mg iron
0.20 mg iron 16.00mL Unknown 1

0.25 mg iron 20.00mL Unknown 2

CALCULATIONS:
Calculating the unknown concentration with help of the beer law:

AR= AU

Where R is termed as the reference and U is the unknown

cU=(IR,cR)/IU

cU3 = (4.8cm ×1.0mg/L)= 0.68mg/L

0.8% U = U3

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 U = 0.68mg/L / 0.8% = 85mgFe3+/L
A3,4 = 0.209 C3,4 – 0.005
C3,4 = (0.135 + 0.005) / 0.209
C3,4 = 0.67mg/L
0.8% U = U3,4
U = 0.68mg/L / 0.8% = 84mgFe3+/L

Unknown iron sample = 110.8mg unknown/50mL = 2216mg/L

U = 85mg Fe3+/L
%Fe3+ = 85/2216 = 3.8% Fe3+

Unknown iron sample = 110.8mg unknown/50mL = 2216mg/L

U = 84mg Fe3+/L
%Fe3+ = 84/2216 = 3.8% Fe3+
mg fe 1000 mL 100 mg
0.10× ×50(dilution factor)× =
50 ml L L
Experimental error−accepted Va ;ue
Relative error= × 100
Accepted Value

Fig 2: Calibration Curve

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RESULT:
A colored complex ion is formed by reaction of iron with o-phenanthroline. Spectroptometer was
used to measure the intensity of colored species. Absorbance versus the concentration curve, a
calibration curve, is obtained. The concentration of unknown amount of iron in the sample was
determined.

CONCLUSION:
The analysis that we used help us to determine the amount of iron in a particular sample. That
calculations were performed accordingly. The iron determination is quite important and it is done
with help of a blank. This is what we use. We do various comparisons and other techniques. A
calibration curve is obtained which is plotted with help of values that is sketched accordingly.
Calibration method is quite important in this regard.

REFERENCES:
Skoog, West, Holler: Fundamentals of Analytical Chemistry (1996) pp. 614-638.
Sawyer, Heineman, & Beebe: Chemistry Experiments for Instrumental Methods (1984)
pp. 242-253.
"General Information" section of manufacturer's Analytical Methods book.
Dietary Reference Intakes: Elements. National Academy of Sciences, Institute of
Medicine,Food and Nutrition Board.
Hussain F, Arayne MS, Sultana NAJMA. Spectrophotometric method for quantitative
determination of iron (III) from iron polymaltose complex in pharmaceutical
formulations. Pak J Pharma Sci 2006;19:299-303.
Thabit Al-Ghabsha S, Amal Saeed M, Usra Al-Neaimy I. Spectrophotometric assay of
iron (II) in pharmaceutical formulation using Alizarin Red Sulphonate reagent. J Edu Sci
2012;25:1-10.
Merli D, Profumo A, Dossi C. An analytical method for Fe (II) and Fe (III) determination
in pharmaceutical grade iron sucrose complex and sodium ferric gluconate complex. J
Pharm Anal 2012;2:450-3.