Experiment 3: Determination of iron in cereal by atomic absorption spectrophotometry

Objective
To determine the amount of iron in cereal samples by flame atomic absorption spectroscopy
using standard addition method.

Introduction
The importance of iron in cereal is to prevent anemia from iron deficiency. This is because the
role of iron in human body are transport oxygen and electrons in respiration process. Red blood
cells consist of iron that enable adequate amount of oxygen to be transported through the whole
body from the lung. It is also increases our human immunity towards certain diseases. If human
do not have a balance diet with foods containing iron, red blood cells will not be able to transport
oxygen efficiently.

The 4 components in atomic absorption spectrophotometry are hollow cathode lamp, burner
system, monochromator and detector. Hollow cathode lamp refers to the light source needed in
order to determine the specific element with different element of hollow cathode lamp. Same
metal was used to make cathode to identified the sample. The materials of making a lamp is a
glass with quartz windows and inert gas(argon) was filled inside of the lamp. The use of
nebulizer in burner system is to reduce the sample in liquid form to a fine aerosol, whereas a
spray chamber and a burned head were used to initiate a flame to produce atoms of the same
element from the sample. Monochromator spreads the incident of the light beam and allows
certain wavelength to reach the detector. Photomultiplier tube was normally used in the detector
to release a signal proportional to the amount of light received by it. Diagram below shows how
the flame-atomic absorption spectroscopy works on its function.

Apparatus and Material

Iron(II) ammonium sulfate hexahydrate, cereal, nitric acid, perchloric acid, flame-AAS

Methodology
0.01g(10mg) of iron(II) ammonium sulfate hexahydrate was dissolved in 100mL of water. 2g of
cereal was ground into powder by using mortar and pestle. 0.5g of cereal was taken and
transferred into a 50 mL beaker. Then, the beaker was placed on a hot plate. 9mL of nitric acid
and 3mL of perchloric acid were mixed and 10mL of this mixture was added into the beaker. The
beaker was gently warmed until the sample appear colourless. After the digestion completed, the
solution is transferred to a 25mL volumetric flask and diluted to the mark with distilled water.
The solution was then filtered.
5 test tubes were prepared, there were blank, 1, 2, 3 and 4. 4mL of the digested sample solution
was added to each test tube. 0.20mL, 0.30mL, 0.40mL and 0.60ml of stock iron(II) solution were
added to test tubes 1, 2, 3 and 4 respectively. Next, 1.0mL of distilled water was added to the
blank test tube and the balance were diluted with distilled water until 5mL of final solution were
formed respectively. All the standard solution were used for flame-AAS analysis.

Results and Calculation

10mg in 100mL 100mg in 1000mL(1L)
Hence, original stock concentration is 100mg/L(=100g/mL)(=100ppm)
Exact concentration of iron:
100ppm x

55.847 g mol1
392.14 g mol1

= 14.2416ppm

Dilution:
M1V1 = M2V2
Where M1 is concentration of original stock concentration, V1 is volume of original stock
concentration
M2 is concentration of iron after diluted, V2 is volume of iron after diluted
Test tube 1: (14.2416ppm)(0.2mL) = M2(5mL)
M2 = 0.569664ppm
Test tube 2: (14.2416ppm)(0.3mL) = M2(5mL)
M2 = 0.854496ppm
Test tube 3: (14.2416ppm)(0.4mL) = M2(5mL)
M2 = 1.139328ppm
Test tube 4: (14.2416ppm)(0.6mL) = M2(5mL)
M2 = 1.708992ppm

Flame-AAS results:
Test tube

Iron concentration, ppm

Absorbance, A

1
2
3
4

0.569664
0.854496
1.139328
1.708992

0.113
0.124
0.148
0.154

Graph of absorbance, A against Iron concentration, ppm

0.18

0.16
f(x) = 0.04x + 0.1

0.14

0.12

0.1

Absorbance, A
0.08

0.06

0.04

0.02

0
0.4

0.6

0.8

1.2

Concentration, ppm

1.4

1.6

1.8

y = 0.0372x + 0.095
Find x-intercept, y = 0
0 = 0.0372x + 0.095
x = -2.5538ppm
Because it is a distance from origin, the negative can be ignored.
x = 2.5538ppm
1mL 2.5538g
25mL 63.845g
Hence, 63.845g Fe in 0.50g of cereal
63.845g x

100 g
0.5 g

= 12769g Fe

= 12.769mg Fe in 100g cereal

Based on information on the cereal box, there is 12mg Fe in 100g cereal.

Discussion
The method of standard addition is a type of quantitative analysis approach often used in
analytical chemistry whereby the standard is added directly to the aliquots of analyzed sample. Is
a technique used to minimize matrix effects that interfere with analyte measurement signals. The
unknown component concentrations are often elucidated through a range of analytical
techniques, such as light spectroscopy, mass spectrometry, and electrochemistry. However, the
measurement can be affected by other components in the sample, called the matrix, and cause the
inadvertent reduction or enhancement of the signal, called matrix effects. Thus making it
impossible to compare the analytical signal between sample and standard using the traditional
calibration curve. Standard addition is frequently used in chemical instrumental analysis such as
atomic absorption spectroscopy and gas chromatography.

In order to determine the percent recovery, it was necessary to calculate the theoretical amount of
iron expected in the cereal sample. From the nutritional information on the package, a 100g
serving of cereal contains 12 mg of Fe. The mass of cereal used for the experimental data was
0.5g which should contain 0.06mg Fe. The Fe contain calculated based on the data given in 0.5g
of cereal is 0.063845mg which means 12.769mg Fe in 100g cereal. When the experimental
amount of Fe (12.769mg) is compared to the theoretical amount (12mg), there are 0.0769mg
(6.4%) of Fe is more than the theoretical amount.