NAME: MOAKOFI

SURNAME: MATSEBE
ID: 201902992

DATE: 03/11/2022

EXPERIMANT 7: THE DETERMINATION OF IN FOOD STUFF BY AAS

TITLE: THE DETERMINATION OF IRONIN FOOD STUFF BY AAS

ABSTRACT
Atomic absorption spectrometry was used to determine the iron content in cereals, beer , coke
and liver. The concentration and percentage of iron in liver sample was determined to be 2.31
mg/L and 2.42% respectively, the concentration and percentage of iron in cereal sample was also
determined and was found to be 0.914 mg/L with a percentage of 0.20% respectively, that of
beer 2.31 mg/lwith a percentage of 1.16 and coke samples concentration was 0.025mg/l and a
percet of 0.00125%.

Introduction

Atomic Absorption Spectroscopy is an instrumental analysis technique for rapid trace metal
analysis. It is based on element specific wavelength light absorption by ground state atoms in the
flame or electro-thermal graphite furnace. It finds immense applications in the analysis for trace
metals in soils, lakes, rivers, oceans, and drinking water, pharmaceuticals, foods and beverages,
geological and mineralogical samples, petroleum products, biological fluids and specimens and
forensic analysis. It is common to get results in ppm levels and a higher sensitivity of ppb levels
when we using graphite furnace atomization. Atomic absorption spectroscopy (AAS) is based
upon the principle that free atoms in the ground state can absorb light of a certain wavelength.
These very specific wavelengths give the technique excellent specificity and detection limits in
the AAS analysis. Absorption for each element is specific, no other elements absorb this
wavelength (Ferreira 2018). These atoms will absorb electromagnetic radiation at a particular
wavelength during the atomic absorption spectroscopy procedure. This generates a discernible
signal claim that it is therefore able to ascertain the parts per million, or ppm, amounts of
specified metals in the substance being tested by taking a look at these signals. There are
electrons in an atom with different energies. The process of spectroscopy involves the absorption
of energy, which causes electrons to migrate to a more energetic level. The transition that takes
place during this phase is closely related to the radiant energy the electrons absorb. When the
atoms are energized, they can absorb light. Light at a resonant wavelength that travels through an
atom cloud is measured by atomic absorption gets taken in by them. Photons are released as
energy when the excited electrons begin to relax once more. Every element has a distinct
electrical structure of their own. Therefore, each element's distinct characteristic is represented
by the radiation that was absorbed (Farey et al2011). By measuring the amount of light absorbed
and the energy emitted during the spectroscopy process, the amount of a specific element that is
present in a substance is identified. In a readout, the variations in these light wavelengths before
and after absorption will show up as energy absorption peaks. Metal content in materials and
liquids can be determined using atomic absorption spectroscopy.
Some of the advantages of AAS include cheapness and comparatively easy and simple to
manipulate the machine; sensitivity such that many element can be determined at ppm level or
even less; high precision and accuracy obtained by the calibration curves; absorption signal
considerably free from inter-element interferences and above all, Richard et al (1993)atoms of a
metal absorb at a well-defined wavelength and over a narrow bandwidth hence isotopes of the
same element will not absorb at each other radiation. Some of the disadvantages include limited
application as only about 70 elements excluding earth metals have been detected by this method.
Another disadvantage in that AAS cannot yet detect non metals.

Iron is a mineral that is naturally present in many foods, added to some food products, and
available as a dietary supplement. Iron is an essential component of hemoglobin, an erythrocyte
(red blood cell) protein that transfers oxygen from the lungs to the tissues (Bacon et al 2011). As a
component of myoglobin, another protein that provides oxygen, iron supports muscle metabolism
and healthy connective tissue. Iron is also necessary for physical growth, neurological development,
cellular functioning, and synthesis of some hormones. Dietary iron has two main forms: heme and
nonheme. Plants and iron-fortified foods contain nonheme iron only, whereas meat, seafood, and
poultry contain both heme and nonheme iron. Heme iron, which is formed when iron combines
with protoporphyrin IX, contributes about 10% to 15% of total iron intakes in western. he richest
sources of heme iron in the diet include lean meat and seafood (Mei et al 2006). Dietary sources of
nonheme iron include nuts, beans, vegetables, and fortified grain products. In the United States,
about half of dietary iron comes from bread, cereal, and other grain products . Breast milk contains
highly bioavailable iron but in amounts that are not sufficient to meet the needs of infants older
than 4 to 6 months Organ meats are good sources of iron, and liver contains 36% of the DV
per serving. Organ meats are also rich in many other nutrients, such as selenium, vitamin A,
and choline. In other countries, wheat and other flours are fortified with iron. Infant formulas are
fortified with 12 mg iron per liter. Heme iron has higher bioavailability than nonheme iron, and
other dietary components have less effect on the bioavailability of heme than nonheme iron. The
bioavailability of iron is approximately 14% to 18% from mixed diets that include substantial
amounts of meat, seafood, and vitamin C (ascorbic acid, which enhances the bioavailability of
nonheme iron) and 5% to 12% from vegetarian diets (Fleming et al 2001). In addition to ascorbic
acid, meat, poultry, and seafood can enhance nonheme iron absorption, whereas phytate (present
in grains and beans) and certain polyphenols in some non-animal foods (such as cereals and
legumes) have the opposite effect. Unlike other inhibitors of iron absorption, calcium might
reduce the bioavailability of both nonheme and heme iron. However, the effects of enhancers and
inhibitors of iron absorption are attenuated by a typical mixed western diet, so they have little
effect on most people’s iron status

Atomic absorption spectroscopy has limitations or is disadvantageous to a certain extent. It uses hollow
cathode lamps during analysis of samples. Lamps are costly and they need to replacement after a certain
period of time. AAS also is affected by chemical interferences since because of its low temperature, some
chemical bonds of metals cannot be broken.
Beer-Lambert's law for absorption spectroscopy is a linear relationship between the absorbance and the
concentration of an absorbing species. The states imply that type, as well as the concentration of the
molecules, are necessary. Beer’s law, also called Lambert-Beer law or Beer-Lambert law, in
spectroscopy, a relation concerning the absorption of radiant energy by an absorbing medium. It states
that the absorptive capacity of a dissolved substance is directly proportional to its concentration in a
solution (welz 2016). The relationship can be expressed as;

A = εbc

Where: A is absorbance

- Ε is the molar absorptivity (depends on the nature of chemical and wavelength of light)

-b is the length of the path light must travel (in the solution in centimeters)

-c is the concentration of a given solution

The aim of this experiment is to use Atomic Spectroscopy to determine iron in liver, cornflakes,
coka cola and beer.
LIST OF REAGENTS

TABLE A: LIST OF REAGENTS AND APPARATUS USED IN THE EXPERIMENT

LIST OF REAGENTS APPARATUS

 H2SO4  Beakers
 FeCl2.6H20  volumetric flaks
 HNO3  Pipettes
 nitric acid  Funnels
 high purity iron granules  Hot plate
 Concentrated HCL  Filter papers
 Distilled water  Conical flasks

PROCEDURE

All needed apparatus were collected. Exactly 0.50 g of high purity iron was weighed and placed
into a 100ml beaker. It was then dissolve in a minimum quantity of 1:1 HCL/water, covered with
a watch class and was then heated to aid dissolution. The solution was cooled then transferred
into a 500.0 ml volumetric flask and diluted to the mark with distilled water. An iron standard of
2 ppm and 0.10 M with respect to HCL was prepared. Then a series of five more standard
(1,2,4.6 and 8ppm) was also prepared. To 20 ml of beer in 100 ml measuring cylinder , 20 ml of
distilled water was added and then the content of the measuring cylinder was poured into a 100
ml beaker. The solution was stirred well to assist the expulsion of carbon dioxide. To 20 ml of
coke in 100 ml measuring cylinder, 80ml of distilled water was added and then the content of the
measuring cylinder was also poured into a 100 ml beaker. The solution was stirred well to assist
the expulsion of carbon dioxide. To 1g of cereal 20 ml of concentrated HNO3 acid was added
and then heated in a fume hood to help digest the cereal, then 0.5 ml of the solution was diluted
in a 10 ml volumetric flask. To 10g of liver, 20ml of concentrated HNO3 acid and 4ml of H2SO4
was added and then heated in a fume hood to help digest the liver until a white smoke of sulfuric
acid appeared, then 0.5 ml of the solution was diluted in a 10 ml volumetric flask . Atomic
absorption spectrometer was adjusted according to the manufactures instruction for the
determination of iron, the hollow cathode lamp emission line at 248.3 nm was selected. Distilled
water was aspirated into the flame to zero the absorbance scale. Then each standard solution and
sample solutions were aspirates in turns, they were rised in between with distilled water and the
absorbance was recorded in each case.
RESULTS AND ANALYSIS
TABLE1: MASS OF SAMPLE SOLUTIONS
SAMPLE MASS
Liver 10.0020grams
Corn flakes 1.0003gram
Coke 20g
Beer 20g

TABLE 2: CONCENTRATION AND ABSORBANCE OF SANDARD SOLUTION Fe

STANDARD CONCENTRATION %RELATIVE MEAN
(mg/L) STARNDARD ABSORBANCE
DEVIATION
Cal zero 0.000 >100 -0.0005
Standard 1 2.000 1.4 0.0209
Standard 2 4.000 0.7 0.0429
Standard 3 6.000 0.5 0.0675
Standard 4 8.000 0.3 0.0813

TABLE 2: CONCENTRATION AND ABSORBANCE OF SAMPLES

Sample Concentration(mg/L) Mean absorbance
Cereal 0.912 0.0096
Liver 2.374 0.0255
Coke 0.025 0.0003
Beer 2.264 0.0243

SAMPLE CALCULATION
(A) Volumes of Ca Standard series
i)For 2ppm
C1V1=C2V2
V1= C2V2/ C1
=2×100ML/100ppm
=2ml
(B)Volumes of Fe Standard series
i)For 4ppm
C1V1=C2V2
V1= C2V2/ C1
=4ppm×100ML/100ppm
=4ml
(C)Volumes of Mn Standard series
i)For 6ppm
C1V1=C2V2
V1= C2V2/ C1
= 6ppm×100ML/100ppm
 =6ml

i)For 8ppm
C1V1=C2V2
V1= C2V2/ C1
=8ppm×100ML/100ppm
=8ml
ACTUAL CONCENTARTION CALCULATION FOR SAMPLES
From the calibration curve
Y=mx+c
Y=0.0105x + 0.0004
Y=absorbance
X=concentration
m=molar absorptivity
Beer Lambert`s Law
A=Ɛbc
A=Ɛ(1)c
Y=mx
Calculation of fe in liver
Y=mx
0.0255=0.0105x
x=0.0255/0.0105
=2.42mg/L
Calculation of fe in cereal
Y=mx
0.0096=0.0105x
x=0.0096/0.0105
=0.914mg/L
Calculation of fe in coke
Y=mx
0.0003=0.0105x
x=0.0003/0.0105
=0.0029mg/L
Calculation of fe in beer
Y=mx
0.0243=0.0105x
x=0.0243/0.0105
=2.31mg/L

Mass of fe in liver
Mass= Concentration × Volume
=2.42×0.1L
0.24g
% of Ca=(Mass of Ca/Mass of sample)×100
=0.24/10.0g*100
=2.42 %
Mass of fe in cereal
Mass= Concentration × Volume
=0.914×0.1L*100
0.0914g
% of fe in cereal=(Mass of fe/Mass of sample)×100
=0.0914/1g*/100
=9.14%
Mass of Fe in Coke
Mass= Concentration × Volume
=0.025×0.1L
0.0025g
% of Fe=(Mass of Fe/Mass of sample)×100
=0.0025/20g*100
 =0.0125%

Mass of Fe in beer
Mass= Concentration × Volume
=2.31×0.1L
=0.231g
% of Fe=(Mass of Fe/Mass of sample)×100
=0.231/20g*100
=1.16%
 GRAPH OF ABSORBANCE OF Fe STANDARDS AGAINST
CONCENTRATION (mg/L)
0.09

f(x) = 0.01051 x + 0.000379999999999998

R² = 0.993764462021786
0.08

0.07

0.06

0.05
Absorbance

0.04

0.03

0.02

0.01

0
0 1 2 3 4 5 6 7 8 9

-0.01

Concentration
DI
SCUSSION
Atomic absorption spectrometry (AAS) well detected elements in liquid samples through its
capability which is the application of characteristic wavelengths of electromagnetic radiation
from a light source. Individual elements absorbed wavelengths differently, and these absorbance
got measured against standards (Welz 2016). In effect, AAS takes advantage of the different
radiation wavelengths that are absorbed by different atoms. In AAS, analytes are first atomized
so that their characteristic wavelengths are emitted and recorded. Then, during excitation,
electrons move up one energy level in their respective atoms when those atoms absorb a specific
energy. This energy corresponds to a specific wavelength that is characteristic of the element.
Depending on the light wavelength and its intensity, specific elements can be detected and their
concentrations measured. AAS determined iron in liver, cereal, coke and beer. Atomic
absorption spectrometry was used to determine the iron content in cereals, beer , coke and liver.
The concentration and percentage of iron in liver sample was determined to be 2.31 mg/L and
2.42% respectively, the concentration and percentage of iron in cereal sample was also
determined and was found to be 0.914 mg/L with a percentage of 9.14% respectively, that of
beer 2.31 mg/l with a percentage of 1.16 and coke samples concentration was 0.025mg/l and a
percet of 0.00125%. According to the data read by atomic absorption spectrometer, the graph plotted
gave the equation of a straight-line y= 0.0105x + 0.0004. Since absorbance is directly proportional
to concentration, then an increase in concentration increases absorbance because if the
concentration of solution is increased, then there are more molecules that can be atomized when it passes
through the flame to the detector. This was proved with a positive gradient of the graph. R squared known
to be the coefficient of determination tells one how many points fall on the regression line. From the
calibration curve above r-squared was found to be 0.9938 so this means that 99.3% of the variation of y-
values around the mean are explained by the x-values. The regression coefficient (R 2) of the linear
regression should be 0.995 to prove linearity. The value obtained in this experiment was R 2=0.9938 and
according to literature, this indicated that there was a strong linear relationship between the two
parameters and this proves an excellent linearity and this indicates that this experiment was carried out
according to Beer-Lambert’s equation

Reagent contamination, analyst contamination, and absorption by container materials are

possible sources of errors. By using high purity research grade reagents for sample preparation,
disposable gloves, clean lab coats, and clean glassware for creating samples and standards, these
errors can be reduced to a minimum.
Atomic absorption spectroscopy is well known for good accuracy, determination in low concentration
The FAAS machine is relatively cheap and easy to operate. Many elements can be determined at
ppm level or even lower. High precision and accuracy are obtained by the calibration curves. The
absorption signal is largely free from interferences from other elements. And, most importantly,
metal atoms absorb at a well-defined wavelength and over a narrow bandwidth. As a result,
isotopes of the same element will not absorb at each other ra. Limited application as only
roughly 70 people use it is one of the drawbacks.

Iron standards must be prepared in weak acid to ensure solubility and accurate measurement. In
close proximity to the most delicate resonance line, iron has non-absorbed lines. The ratio of the
concentration of atoms to the amount of light absorbed is not constant. This is particularly clear
at greater concentrations, because the resonance emission line's strength is low in comparison to
the intensity of the unabsorbed line. The effect is severe curvature of the calibration graph
towards the concentration axis. It is necessary to expel carbon dioxide from samples to reduce
the influence of dissolved carbon dioxide gas on nebulization and transport. Coke has high iron
content than beer because it had a relative standard deviation which was greater than 100,
indicating too much content of it curve. Liver has low percent because the mass of the sample
was much graeter than cereal that’s why the percentage of fe in cereal is more than in liver. Coke
had least amount of iron.

CONCLUSION
Atomic absorption spectrometry was found to be an efficient technique for determining metal trace
amounts in samples.Atomic Spectroscopy was used to determine iron in liver, cereal, coke and
beer. The concentration and percentage of iron in liver sample was determined to be 2.31 mg/L
and 2.42% respectively, the concentration and percentage of iron in cereal sample was also
determined and was found to be 0.914 mg/L with a percentage of 0.20% respectively, that of
beer 2.31 mg/lwith a percentage of 1.16 and coke samples concentration was 0.025mg/l and a
percet of 0.00125%.
REFERENCES

Bacon BR, Adams PC, Kowdley KV, Powell LW, Tavill AS. Diagnosis and management of
hemochromatosis: 2011 practice guideline by the American Association for the Study of Liver
Diseases. Hepatology 2011;54:328-43. [PubMed abstract]

Farey, H. J. and Nelson, L. A. (2011) ‘Atomic Absorption Spectrometry’, Techniques and

Instrumentation in Analytical Chemistry, 5, pp. 67–94.

Ferreira, S. L. et al., 2018. Atomic Absorption Spectrometry - A Multi-element Technique.

Trends in Analytical Chemistry, Volume 100, pp. 1-6.

Fleming DJ, Jacques PF, Tucker KL, Massaro JM, D’Agostino RB, Sr., Wilson PW, et al. Iron
status of the free-living, elderly Framingham Heart Study cohort: an iron-replete population with
a high prevalence of elevated iron stores. Am J Clin Nutr 2001;73:638-46. [PubMed abstract]

Mei Z, Cogswell ME, Looker AC, Pfeiffer CM, Cusick SE, Lacher DA, et al. Assessment of iron
status in US pregnant women from the National Health and Nutrition Examination Survey
(NHANES), 1999-2006. Am J Clin Nutr 2011;93:1312-20. [PubMed abstract]

Welz, B (2016) Speciation analysis, The future of atomic absorption spectrometry, J. Anal. At.
Spectrom. (13) 413 – 417