I.

Title of Experiment : Analysis Fe Metal in the Water Sample

(wheel) Using AAS Instrument
II. Day, Date of Experiment : Friday, November 24th, 2017
III. Purpose of Experiment : To determine Fe level in the water
sample (wheel)
IV. Basic Theories

Atomic Absorption Spectrometry (AAS) is a technique for measuring

quantities of chemical elements present in environmental samples by
measuring the absorbed radiation by the chemical element of interest. This is
done by reading the spectra produced when the samples excited by radiation.
The atoms absorb ultraviolet or visible light and make transitions to higher
energy levels. Atomic absorption methods measure the amount of energy in
the form of photons of light that are absorbed by the sample. A detector
measures the wavelengths of light transmitted by the sample, and compares
them to the wavelengths which originally passed through the sample. A signal
processor then integrates the changes in wavelength absorbed, which appear
in the readout as peaks of energy absorption at discrete wavelengths. The
energy required for an electron to leave an atom is known as ionization energy
and is specific to each chemical element. When an electron moves from one
energy level to another within the atom, a photon is emitted with energy E.
Atoms of an element emit a characteristic spectral line. Every atom has its
own distinct pattern of wavelengths at which it will absorb energy, due to the
unique configuration of electrons in its outer shell. This enables the qualitative
analysis of a sample. The concentration is calculated based onthe Beer-
Lambert law. Absorbance is directly proportional to the concentration of the
analyte absorbed for the existing set of conditions. The concentration is
usually determined from a calibration curve, obtained using standards of
known concentration. However, applying the Beer-Lambert law directly in
AAS is difficult due to: variations in atomization efficiency from the sample
matrix, non-uniformity of concentration and path length ofanalyte atoms (in
graphite furnace AA) (Gracia and Baez, 2012).
 Well water that is suitable for consumption as drinking water must meet
the water quality standards specified in Government Regulation No. 82 of
2001. One of the standards that must be met for drinking water is the content
of metal ions such as Fe (Iron), Zn (Zinc) and Mn (Manganese) which is
mostly contained in ground water. Water is the most important requirement in
human life, so water quality requirements are important for us to know. Water
that we consume every day must meet the quality of drinking water, which
contains maximum iron 0.3 ppm (Department of Health, 2002). In surface
water rarely encountered iron levels exceeding 1 mg / L, but in groundwater
such as well water the iron content can be much higher, because in the soil
contains many minerals. The iron contained in the well water can be
determined by performing a quantitative analysis using atomic absorption
spectrophotometer (AAS) using an iron cathode hallow lamp.

Metals are natural constituents of the environment. At high

concentrations, trace metals can become toxic for living organisms and
behave as conservative pollutants. Metals enter the environment mainly by
two means: (i) natural processes (for example, erosion of rocks, volcanic
activity, forest fires) and (ii) processes due to human activities. An
anthropogenic activity may add considerable amounts of polluting
compounds, which will influence the existing natural aquatic system. Metals
are frequently released in large quantities during different processes derived
from human activities and may lead to major destruction of aquatic
ecosystems. Moreover, human beings located at places contaminated by heavy
metals could be especially sensitive to these contaminants due to
bioaccumulation. The metals can accumulate in bone, hair and in some soft
tissues, such as the liver, kidney, brain or lungs.

At low concentrations, iron plays an important role in metabolic and

fermentation processes, as an enzyme activator, stabilizer and functional
component of proteins. Above trace levels, however, iron has other roles. It
has been stated that forall trace elements that considered to be essential,
including iron, there exists afairly narrow concentration window between
the essential and toxic levels. Ironis a moderately toxic element when
compared with other transition metals. However,the toxic doses of iron and its
compounds can lead to serious problems, including depression, rapid and
shallow respiration, coma, convulsions and cardiacarrest. Iron and its
compounds have widespread industrial applications (constructional material
for drinking-water pipes, food colours, coagulants in water treatment,
pigments in paints and plastics); hence, large quantities of iron are discharged
into the environment. Thus, appropriate knowledge of the iron content of
natural water is very desirable. So, the toxicity of this element demands a fast
and accurate method for its determination in water (Tautkus et al, 2004).

It is widely accepted that the presence of Fe (III), alone or in

combination has beneficial or deleterious effects on the properties of many
substances and the nature of various biological systems. It provides a
fundamental structure of hemoglobin, myoglobin, hem-enzymes and many co-
factors involved in enzyme activities. Trace amounts of Fe (III) in various
substances may be vital and can promote rancidity. It plays a central role in
the biosphere and serves as the active center of proteins responsible for O 2
transport and electron transfer mechanisms and of metallo-enzymes such as
oxidases, reductases, and dehydrases. Fe (III) may control the mobility and
toxicity of other metals, whilst Fe (III) may also be a limited nutrient for
phytoplankton growth in the open ocean, Fe (II) is probably the preferred
nutrient. The concentration of Fe3+. ions in biological fluids is usually found at
trace level, requiring sensitive instrumental techniques. Frequently a pre-
concentration step is required when a complex matrix has to be analyzed.
Flame atomic absorption spectrometry (FAAS) has been widely used for the
determination of trace metal ions, a relatively simple and precise technique.
However, direct determination of metal ions at trace levels by FAAS is limited
not only due to insufficient sensitivity, but also by matrix interference. For
this reason, preliminary separation and pre-concentration of trace elements
from the matrix is frequently necessary to improve the detection limit and the
selectivity (Ghaedi et al, 2008).This analytical technique is remarkable for its
selectivity, speed and fairly low operational cost (Tautkus et al, 2004).

Source : web.nmsu.edu

Basic Principle of AAS

The selectivity in AAS is very important, since each element has a

different set of energy levels and gives rise to very narrow absorption lines.
Hence, the selection of the monochromator is vital to obtain a linear
calibration curve (Beers' Law), the bandwidth of the absorbing species must
be broader than that of the light source; which is difficult to achieve with
ordinary monochromators. The monochromator is a very important part of an
AA spectrometer because it is used to separate the thousands of lines
generated by all of the elements in a sample. Without a good monochromator,
detection limits are severely compromised. A monochromator is used to select
the specific wavelength of light that is absorbed by the sample and to exclude
other wavelengths. The selection of the specific wavelength of light allows for
the determination of the specific element of interest when it is in the presence
of other elements. The light selected by the monochromator is directed onto a
detector, typically a photomultiplier tube, whose function is to convert the
light signal into an electrical signal proportional to the light intensity. The
challenge of requiring the bandwidth of the absorbing species to be broader
than that of the light source is solved with radiation sources with very narrow
lines(Gracia and Baez, 2012).

The light source in AAS is the light source of the cathode lamp
coming from the element being measured and then passed into a flame
containing the atomized sample, then the radiation is passed to the detector
through the monochromator. Chopper is used to distinguish radiation from a
flame. The detector will reject the direction of the current (DC) of the flame
emission and mnegukur only alternating current from the source of radiation
or sample. The atoms of an element in the basic state will be subjected to
radiation then the atom will absorb energy and cause the electrons in the outer
shell to rise to a higher energy level or excited. The atoms of the sample will
absorb some of the light emitted by the light source. The absorption of light
energy occurs at a certain wavelength corresponding to the energy required by
the atom (Basset, 1994).

If light of a certain wavelength is passed to a cell containing the

corresponding free atoms then some of the light will be absorbed and the
absorption intensity will be proportional to the number of free metal atoms in
the cell. The relationship between absorbance and concentration is derived
from:

1. Lambert's Law:
When a monochromatic light source passes through a transparent
medium, then the intensity of the transmitted light decreases with the
increasing thickness of the absorbing medium.
2. Beer's Law:
 The intensity of the transmitted light decreases exponentially with
the increasing concentration of the species that absorbs it.

The quantitative relationship between the intensity of the absorbed

radiation and the concentration of the elements present in the sample solution
is the basis of SSA usage for the analysis of the metal elements. To form a
neutral atomic vapor in a state / level of basic energy ready to absorb radiation
requires a certain amount of energy. This energy usually comes from the
burning flame of a mixture of acetylene-air or acetylene-N 2 O gas, depending
on the temperature required to make the analyte element into free atomic
vapor at the ground state level. Here is the relationship known as Lambert-
Beer's law which became the basis of SSA's quantitative analysis. The
relationship is formulated in the following equation (Ristina, 2006).

I = Io . a.b.c
Or
Log I/Io = a.b.c
A = a.b.c
with,
A = absorbance, without dimension
a = absorption coefficient,
b = the length of the light trace in the medium containing the absorbing atom,
c = concentration,
Io = the intensity of the first light
I = the intensity of the transmitted beam

In the above equation it is shown that the magnitude of absorbance is

directly proportional to the concentration of atoms at the elementary level in
the flame medium. The amount of concentration of atoms in the flame is
proportional to the element concentration in the sample solution. Thus, from
the absorption plot and the element concentration in the standard solution the
calibration curve is obtained. By placing the absorbance of a sample on the
standard curve a concentration is obtained in the sample solution.
The way to determine the concentration of the sample solution is to
compare the absorbance (A) of the sample solution with the absorbance of the
known standard solution of concentration. Next is made a calibration curve is
a graph of the relationship between absorbance (A) to the concentration of
standard solution in the form of a straight line. The sample solution is
measured for absorbance, then plotted on the calibration curve. Thus the
concentration of the sample can be determined.

Lambert Beer's Legal Curve (calibration curve)

The parts of AAS are as follows (Day, 1986).

1. Resonance radiation source
Resonance radiation source used is hollow cathode lamp (Hollow
Cathode Lamp) or Electrodeless Discharge Tube (EDT). Hollow cathode
lamp electrodes are usually composed of tungsten and hollow cathodes
coated with pure elements or mixtures of desired pure elements. Lamps
and window shutters are made of silica or quartz, filled with a fill gas that
can produce ionization processes. The normally used gas is Ne, Ar or He.
The emission of resonant radiation occurs when both electrodes are given
a voltage, the electric current generating the ionization of the fill gas.
These positively charged gas ions fired at the atoms at the cathode causing
the excitation of the atoms. These excited atoms are unstable and will
 return to the ground level by releasing their exit energy in the form of
radiation. This radiation is passed through the atoms in the flame.
2. Gas cylinders
The gas tube in the AAS used is a gas tube containing acetylene
gas. Acetylene gas in AAS has a temperature range of 20000K, and there
is also a gas tube containing N2O gas hotter than acetylene gas, with a
temperature range of 30000K. the regulator on the acetylene gas tube
serves to regulate the amount of gas to be released, and the gas inside the
tube. Spedometer on the right side of the regulator. Is a pressure regulator
inside the tube. Testing for the detection of leaking or not the gas cylinder,
namely by closing the ear to the near gas regulator and given a little water,
to check. When a sound or air is heard, it is suspended that the gas
cylinder is leaking, and there is a gas coming out. Another thing that can
be done is to provide a little soap water at the top of the regulator and see
if there is a bubble of air that is formed. If there is, then the gas cylinder is
positively leaked.
We recommend checking the leak, do not use oil, because the oil
will be able to cause clogged gas. The gas inside the tube can come out
because it is caused inside the tube at the bottom of the tube containing
acetone that can make the gas will easily come out, in addition to gas also
has pressure.
3. Ducting
Ducting is part of the chimney to suck smoke or burning residue
on the AAS, which is directly connected to the exterior chimney on the
roof of the building, so that the smoke produced by AAS, not harmful to
the environment. Smoke generated from combustion in AAS, is processed
in such a way in ducting, so that the resulting ppolution is harmless.
How to maintain ducting, ie by closing the ducting horizontally, so
that the top can be tightly closed, so there will be no insects or other
animals that can enter into the ducting. Because if there are insects or
other animals that enter into the ducting, it can cause ducting congestion.
 The use of ducting is, pressing the small part of the ducting
towards the incline, because when horizontally straight, indicating ducting
closed. Ducting serves to suck the results of the culprit that occurs in the
AAS, and remove it through a chimney that is connected with ducting.
4. Compressor
The compressor is a separate device with the main unit, as it serves
to supply the air requirements to be used by AAS, at the time of atomic
combustion. The compressor has three pressure control knobs, whereas in
the black box the ON-OFF button, the center spedo is the size of the air to
be removed, or the pressure regulator, while the right button is the
adjustment key to adjust the amount of air will be sprayed onto the burner.
The sections on the back of the compressor are used as air storage
after the use of AAS. This tool serves to filter the air from the outside, so
clean.position to the right, is the open position, and the position to the left
meerupakan closed position. The water vapor released will splash and can
cause the surrounding floor to become wet, so it is advisable to press on to
the right of this section, preferably accommodated with a cloth, so that the
floor does not get wet, and moisture will be absorbed into the cloth.
5. Atomizer
Atomizer consists of Nebulizer (spray system), spray chamber and
burner (burner system). The nebulizer serves to convert the solution into
an aerosol (fog grains with particle size 15 - 20 m) by pulling the
solution through the capillaries (due to the effects of airflow) by
suctioning the fuel gas and oxidants, sprayed into the atomizer chamber.
The fine mist particles then together the gas fuel mixture flow into the
flame, while the large mist point is drained through the drain. Spray
chamber serves to create a homogeneous mixture of oxidant gas, fuel and
aerosol containing the sample before entering the burner. Burner is an
exact system of atomization occurring that is changing the fog / vapor of
the elemental salt to be analyzed into normal atoms in flame. . Chopper is
 used to distinguish radiation from a source of radiation, and radiation from
a flame.
6. Monochromator
After the resonant radiation from the hollow cathode lamp through
the atomic population inside the flame, this radiation energy is partly
absorbed and partly passed on. The radiation fraction passed is separated
from other radiation. The choice or separation of radiation is performed by
a monochromator.
The monochromator serves to separate the radiation of the
resonance that has undergone such absorption from other radiations. Other
radiation comes from hollow cathode lamps, hollow cathode gas filler gas
or impurity metal in hollow cathode lamps. Monochromator consists of
optical system that is gap, mirror and lattice.
7. Detector
The detector measures the radiation transmitted by the sample and
measures the intensity of the radiation in the form of electrical energy.
8. Recorder
The electrical signal coming out of the detector is received by a
device that can automatically represent the absorption curve.

Interference in AAS

Interference is defined as an effect of the matrix component on the

results of the analysis. Disturbance leads to differences in behavior in the
sample and calibration solution. Disorders can be divided into two classes:
spectral disorders and non-spectra disorders.

1. Spectral Interference,

causing an increase in absorption:

 a) Background Spectral Caused by scattering particles in atomization or
molecular absorption, partly due to the difficulty of breaking the
oxides, hydroxides or halides. Can be overcome using D2 lights.

b) The presence of of other elements very close to the analytes as

follows:

Cd 288,802 nm disturbed As 288,812 nm

Mg 285.213 nm disturbed Fe 285,179 nm

Zn 213,856 nm disturbed Fe 213, 859 nm and Cu 213,850 nm

The disturbance is difficult to remove, the way to overcome it is to

make measurements on other , although usually will give less sensitive
results.

2. Nonspectral (Nonspectral interference)

causing an increase or decrease in absorption.

a) Transport disturbance

Also commonly referred to as a physical disorder, because the

causes of this type of disorder are physical properties (surface tension,
viscosity and specific gravity). These properties influence from the
suction process on the capillary pipe, the formation of aerosol and its
drainage into the flame. Organic solvents give a positive effect (greater
yield) due to its specific gravity, surface tension and viscosity than
water. The lower surface tension will form the finer aerosol grains,
allowing more into the flame. Inorganic salts, inorganic acids and
macro organic molecules (proteins, sugars) will form larger grains so
that the part entering into the flame will be less, this will reduce the
sensitivity and cause negative effects (smaller yield).

b) Ionic disturbances

The presence of atoms from easily ionized elements at the

flame temperature, will cause an equilibrium disorder of the formation
of ions and atoms of the element that is being set, especially when the
concentration of the intruder element is quite large, eg Na. M M+ +
e (example) Na Na+ + e (disturber). Electrons from Na will shift the
first equilibrium to the left. Thus the number of atoms that formed as
bigger, causing the absorption of light will rise and positive error
occurs. To overcome this problem, we use radiation buffer solution
such as CsCl and SrCl2 solution.

c) Emissions Disorder

The free atom can be excited when it absorbs a certain amount

of energy both light energy and thermal energy (flamefotometry), so

that when it returns to its ground state, it releases the emission light.

Because emission light is equal to transmitted light (from HCl),

this type of disturbance can not be removed by the monochromator. To

overcome this disturbance is used modulator. There are 2 types of

modulation systems: a. Electronic modulation By the light modulator

of HCl is made to blink at a certain frequency, so that when received

the detector will produce the current drawing as in Figure 23a, which

is identical with the curve of alternating current. While emission rays

 originating from flame, are continuous rays so that when received the

detector will produce a fixed current strength as in Figure 23b, which

is identical with the direct current curve. By filtration using a device,

the entry into the readout system is simply an alternating current (It),

while direct current (emission light) is eliminated. Electronic

modulation is used in a single beam SSA. By chopper, the light

entering into the flame will be made light dark (retained propeller).

Thus the light transmission (It) becomes dark light. When received by

the detector an alternating current curve will be generated. While that

comes from the emission light generated direct current curve. That is

passed only that.

(Gracia and Baez, 2012)

The advantages and disadvantages of SSA

1. Some advantages of SSA

a. Sensitivity (sensitivity): this way is very concentrated, many elements can
be determined at ppm levels, even some elements with certain techniques
can be determined in order ppb.
b. Selectivity: this way is very selective, so it can determine several elements
at once in a sample trailer without the need for separation.
c. Research and Accuracy: the accuracy of SSA is relatively good because
the disturbances in measurement turn out to be less than other instruments.
The accuracy of the SSA is quite good, because it is simply the cue and
the result of the measurement that is the basis of making the calibration
curve.
2. Some weaknesses of SSA
 a. Some elements do not readily produce atomic vapors in the ground state
when they reach a flame, such as the dissociation of a stable compound
thus blocking detection and determination eg Al, Mo, Si and Ti
b. Some flame is more appropriate for certain elements, so increasing the
sample to be determined requires not only one exchange of light sources
and settings, but also the exchange of flame, burning and gas source.
(Khopkar, 1990)

V. Materials and Equipment

Materials
- Fe Standard solution
- Aquadest
- HNO3 1%
- Well water sample
Equipment
- Beaker glass
- Pipette
- AAS instrument
VI. Procedures of Experiment

1. Standard Solution

Fe Standard Solution 50 ppm

Diluted with aquadest until get 1,2,4 and 6 ppm concentration

Fe Standard Solution 1, 2, 4, 6 ppm

Read absorbance using AAS at 248.3 nm wavelength

Absorbance
2. Blank Solution

Aquadest

Read absorbance using AAS at 248.3 nm wavelength

Absorbance

3. Sample

Water sample

If turbid the solution is added HNO3 1%

Read absorbance using AAS at 248.3 nm wavelength

Absorbance

Calculated Fe concentration

Fe level of sample
 References
Basset, J, et al. 1994. Buku Ajar Vogel; Kimia Analisis Kuantitatif Anorganik.
Penerbit buku kedokteran EGC. Jakarta.

Day, R.A. 1986. Analisa Kimia Kuantitatif. Erlangga: Jakarta.

Departemen Kesehatan (2002). Keputusan Menteri Kesehatan RI No.

907/MENKES/SK/VII/2002 tentang Syarat-syarat dan Pengawasan
Kualitas Air Minum. Jakarta.

Ghaedi, Mehrorang et al. 2008. Combination of Cloud Point Extraction and

Flame Atomic Absorption Spectrometry for Preconcentration and
Determination of Trace Iron in Environmental and Biological
Samples. Central European Journal of Chemistry. Turkey : University
of Erciyes.

Gracia, R; Baez, A. P. 2012. Atomic Absorption Spectrometry (AAS). Mexico

City: University Campus STeP Ri.

Khopkar, S.M. 1990. Konsep Dasar Kimia Analitik. Penerbit Universitas

Indonesia. Jakarta.

Pemerintah Republik Indonesia, 2001. Peraturan Pemerintah Nomor 82

tahun 2001 Tentang Pengelolaan Kualitas Air dan Pengendalian
Pencemaran Air. Jakarta
Ristina, maria. 2006. Petunjuk Praktikum Instrumen Kimia. STTN Batan:
Yogyakarta

Tautkus, Stasys et al. 2004. Determination of Iron in Natural and Mineral

Waters by Flame Atomic Absorption Spectrometry. J.Serb.Chem.Soc.
Naugarduko