Faculty of Medicine

Department of Pharmacy
Toxicology Service CHU
ORAN
2022-2023

Second year course

post graduation

Spectroscopy
of Absorption
Atomic

Presented by: Benabdallah Imene

Supervised by: Pr Rezk-Kallh.H
By Chefirat.B
Dr. Bendjemaa A.
 Atomic Absorption Spectroscopy

Plan:

Introduction

Principle of the SAA

Instrumentation

Interferences and corrections

Applications

Conclusion

1
 Atomic Absorption Spectroscopy

I. INTRODUCTION:

Atomic Spectroscopy (AS) is a spectral analysis technique that studies

the absorptions or emissions of light by the free atom. The elements
Inorganic (metals and non-metals) present in a sample are identified.
and quantified thanks to their atomic spectrum. This is only achievable under the condition
that a significant part of the molecular sample, solid or liquid, be
transformed into atomic gas (vaporization and atomization).
The main techniques involving the SA are:
Atomic flame absorption/emission SAAF/SEAF
Electrothermal Atomic Absorption SAAE
The atomic emission in coupled plasma induced high frequency ICP-ES
The emission of arc or spark
The choice of method will be made according to the type of information one wishes to obtain.
but also depending on the type of sample to be analyzed. Due to the process
, all these methods are destructive. They allow for the analysis of
more than 70 elements in liquid, solid or gaseous phase and the detection limit
can vary between ppm and ppb. Although it is theoretically possible to process all the
elements of the periodic table, technological limitations do not allow
to achieve satisfactory results for hydrogen, nitrogen, oxygen,
halogens and noble gases.
ATOMIC ABSORPTION SPECTROSCOPY (AAS): AAS is a
technique described for the first time by Walsh in 1955. It provides a powerful
quantitative analytical instrument.

II. LASAA PRINCIPLE:

It is the analysis by the absorption of radiation of a certain wavelength (UV-
Vis), by free atoms in their ground state leading to a transition of one of its
external electrons from one electron orbit to another and a change of
energy. This absorption is specific to each element. A source of
radiation emits specific radiations corresponding to the difference
of energy between the ground state and an excited state of the sample to be analyzed.
The analyte must be transformed into free atoms in the ground state capable

2
 Atomic Absorption Spectroscopy

to absorb part of this radiation. The non-absorbed radiation passes through

a monochromator to a detector.
The absorption is then measured; it directly depends on the number of particles.
in the ground state which is linearly dependent on the concentration of the analyte
= , where k is a coefficient specific to each element).

III. INSTRUMENTATION OR EQUIPMENT:

An atomic absorption spectrophotometer generally includes a source,

an atomizer, a monochromator, a detector, and an acquisition device.
There are single beam spectrometers and double beam spectrometers.
Sensitivity is determined by the radiation source and the atomizer. Since
The lamps commonly used have the same line width, the largest.
the difference is attributable to the type of atomizer. Each of the techniques uses
different types of atomizers.

1. Source of radiation:
It is a light source that emits the characteristic spectrum of the element at
analyze. It is important that the source emits lines that are narrower than the line
of absorption, thus three types of lamps meet this requirement:
a. Hollow Cathode Lamps (HCL):
The principle of operation is always the same, the lamps are made up of:
From a glass tube about twenty centimeters long and 3 to 5 cm in diameter.
diameter, closed at the end by a quartz window transparent to UV
From a tungsten (W) anode generally;
From a cylindrical cathode in the shape of a small cup of about 1 cm
depth and 3 to 5 mm in diameter. The bottom of the cathode, which is the
the essential part of the lamp is machined into the element that one wants to measure, or
covered with the element or an alloy of it.
The air has been evacuated from the lamp and replaced with an inert gas, Neon or Argon.
under low pressure. (0.5 to 1.3 kPa).
A potential difference between 100 and 400 V is applied between
the anode and the cathode. A discharge will occur between the two electrodes and this
discharge will be concentrated in the cathode which will emit high-energy electrons.
3
 Atomic Absorption Spectroscopy

energetic. These electrons will collide with the atoms of the gas of
filling and will ionize it.
The Ar+ or Ne+ ions will then be attracted to the cathode by undergoing a
significant acceleration. The acceleration will give them such kinetic energy
that, by striking the bottom of the cathode, they will tear an atom from it.
The metal vapors confined in the cathode are subjected to an intense flow.
argon ions (or neon) as well as electrons. This results in a large number of
collisions resulting in the transition to the excited state of atoms
metallic. From the excited state, they will return to the ground state by emitting
each one a photon whose energy will depend on the excited state to which the atom has been.
carried.
The light emitted by the lamp is therefore not monochromatic, but is
composed of several characteristic lines of the element. Among these, the most
intense will be the one that corresponds to the most probable transition, that is to say the one
which involves the first excited level, at the wavelengthλ0.
b. Electrode-less discharge lamp:
It consists of a sealed quartz tube containing an inert gas and a small
amount of a metallic species, or one of its salts, all under a pressure of
a few mmHg. The energy is supplied by an intense electrostatic field. The
inert gas is ionized and the ions are accelerated to an energy necessary for
to extract and excite the metal atoms. There is then, just like for the lamp at
hollow cathode, a characteristic radiative de-excitation.
The HCL and EDL lamps are perfectly complementary, the former
providing excellent results for non-volatile metals, the seconds
addressing particularly well the study of volatile metals (As, Bi, Ge, Pb, Sb,
Se, Sn).
c. Metal vapor lamps
They use the light emission produced by an electrical discharge in a
gas under pression reduced.
This lamp contains the metallic element inside. There is an anode and a
cathode; the metallic element must be volatilizable at heat (e.g.: Hg, Na).
Mercury vapor discharge lamps, for example, are used in the
atomic absorption devices that are dedicated to the measurement of this metal.

4
 Atomic Absorption Spectroscopy

2. The atomizer:
The SAA requires having the atoms in their ground state in order to observe the lines.
characteristics of the element. The atomizer must therefore provide free atoms without
heat it up. It takes heat to pass the sample usually in
solution to the state of an atomic gas. This heat can be generated by a flame
or by a graphite furnace.
The flame SAA only analyzes the solutions, while the furnace SAA of
graphite analyzes solutions, liquid sludges, and solids.
a. A flame :
The element is present in the form of chemical compounds in a solution.
(aqueous or organic) at a certain concentration. Its introduction into the
flame is created by spraying into the oxidizing gas, using, the gas passes
a narrow opening, at the exit of which it acquires a very high speed; this results in
at this place a significant decrease in pressure. If one places the end there
from a tube plunging into the solution, it is drawn at a constant flow rate, and it
broken down in the gas jet into droplets of variable diameter, but centered
around a value of the order of a few tens of microns. The jet charged with
Droplets traverse a container on the walls of which the large ones settle.
droplets (which would create instabilities in the flame), then is sent to the
burner.
The number N of atoms sent into the flame in the form of combination
chemical is proportional to the concentration of the solution, the factor of
proportionality depending on a large number of factors: speed of the gas jet,
viscosity of the solution, etc.
The flame is produced using a burner powered by a mixture
combustible/oxidizer. Ex: acetylene/air
We distinguish two types:
Turbulent flame burner = diffusion burner.
The fuel and the oxidizer are brought separately: it is essential.
when we have gas mixtures that may explode
(oxygen/hydrogen).
Disadvantage: unstable flames.
Laminar flame burner = pre-mix burner:

5
 Atomic Absorption Spectroscopy

used only in recent devices.

The gases are mixed before arriving in the flame; there is a chamber of
pre-mix.
The flame is much more stable due to the pre-mixing.

Gas mixture
A fuel and an oxidizer are required. The gas mixture is chosen based on
the temperature necessary to obtain the atom in the vapor state.
Example: Na (1800 °C)⟹ butane or propane
Al (over 2000 °C)⟹ hydrogen

Fuel/oxidizer mixture Max T°C

butane/air 1700 – 1900 °C

propane/air 1915°C

hydrogen/air 2100 °C

hydrogen/oxygen 2850 °C

acetylene/air 2125 – 2400 °C

acetylene/nitrous oxide (N2O)2O) 2950°C

b. Four graphite :
Atomization occurs in a cylindrical graphite furnace, open at both ends.
ends and which contains a hole in the center for the presentation of samples.
Two flows of inert gas (argon) are used. The external flow prevents
the air entering the home and the internal current ensures that the vapors generated
in the sample matrix are quickly removed from the oven. This
atomization occurs in three stages; evaporation, decomposition, and finally

6
 Atomic Absorption Spectroscopy

the actual atomization. The heating of the sample causes evaporation

and the decomposition, thanks to the contact of the tube with electrical parts,
through which a more or less intense current is passed, which has the effect of
effect of raising the temperature (2000 to 3000 °C) and thus allows for atomization
the sample in a few milliseconds.
The temperature rise is less abrupt.
and atomization is delayed. In order to improve
reproducibility, we often introduce a
platform inside the tube in such a way
that the sample is no longer in contact with
him. Finally, the absorption spectra are
collected through an acquisition device
high-speed data, the latter
proving necessary as the products
Atomization escapes quickly from the tube.
Cooling and safety systems
inert gas flows must be
previously set up.

c. Atomization of hydrides
Certain elements: As, Pb, Se, Sb, Bi are very difficult to measure using a
four in graphite. Indeed, regardless of the matrix, it is practically impossible
to decompose the sample without losing the analyte, given its high volatility.

7
 Atomic Absorption Spectroscopy

These elements are mainly As, Ge, Se, Te, to which one must also add the
mercury, although its case is treated differently. The five elements that we
We have just mentioned, as well as Bi, Pb, Sb, and Sn, have the property of forming hydrides.
Volatiles. All these compounds are gases at room temperature.
The installation consists of two parts:
The hydride generator itself which is located outside of
spectrophotometer.
The atomization cell that is placed in the optical beam, in place of
the flame or the oven.
A sample (from 1 to 30 mL), acidified with HCl, is placed in a flask.
of reaction. Initially, the installation is purged by a gas current.
inert so as to eliminate the oxygen that would destroy the hydrides. From NaBH4(in
KOH at 2% is then introduced into the solution to be analyzed. It is found in
acidic environment and decomposes rapidly according to the reaction:
NaBH4 + 3H2O + HCl⟶H3BO3 + 8H−
As3+ → NaBH4/H+ AsH3 → 800∘CAs+3/2H2
At the time of decomposition, there is the formation of radical hydrogen.
a lot more reactive what hydrogen molecular.
It is this radical hydrogen that contributes to the formation of hydrides.
The formed hydride is carried by the gas flow towards the atomization cell.
This consists of a long quartz tube about fifteen centimeters long.
long, placed, most often, in an electrically heated oven at temperature
constant of 900 to 1,000 °C. This temperature is sufficient to decompose the
hydrides and release the atoms. The tube is illuminated by the primary source of
the element to be dosed and the absorbance signal is measured in a conventional manner.
There are currently fully automatic generation systems.
of hydrides.

8
 Atomic Absorption Spectroscopy

d. Cold steam atomization

Mercury represents a completely unique case in SAA. Indeed, during
during the reduction step, metallic mercury is formed directly in the state
atomic. The vapor pressure of mercury is sufficient, even at temperature
ambient, so that he can break free. That is why we call it
this technique absorption atomic à vapors cold.
The reducing agent can be NaBH.4or even SnCl2, this one acting according to the
reaction :
Sample: H_2SO_4/ HNO_3, Cl_2SnH2SO4/HNO3, Cl2Sn
HgCl2 + SnCl2⟶Hg+SnCl4
This method has led manufacturers to design completely new devices.
dedicated to this dosage.

9
 Atomic Absorption Spectroscopy

3. The monochromator:
In general, it is not necessary to use a high-precision monochromator.
because the width of the line of the source is a first selection. A simple filter of
glass is often suitable for some alkali metals. However, most of the
SAA instruments are equipped with a monochromator. Its role is to choose
the most intense stripe and to eliminate all light, whatever its source, having
a wavelength different from the one we are working with (the lines of the gas of
filling in the source, any impurities or the atomizer.
4. The detector:
It measures the light intensities necessary for calculating absorbances.
The specific absorption is due to the element to be measured. The non-specific absorption is
due to the continuous absorption of the matrix. We have: Specific absorbance =
Total absorbance - Non-specific absorbance Several types of detectors are
adequate. The choice of this will be made for each selected absorption line
for the analysis, based on its frequency response. The most sensitive detector
commonly used is a photomultiplier.

IV. INTERFERENCES AND CORRECTION:

We call disturbance, interaction, or interference the influence of one or
several constituents of the analyzed medium on the dosage of an element.
The concentration of the analyte determined without considering an interaction is
called apparent concentration; this can be higher than the concentration
real, there is then exhilaration; otherwise, depression. The disturbances
in SAA are classified in three ways:
10
 Atomic Absorption Spectroscopy

Spectral interferences
Chemical interferences
Physical interferences
Ionic interferences

Spectral interferences:
These interferences affect the spectral absorbance measurement of the analyte in
causing errors in the determination of its concentration by:
Superposition of the line of the element to be measured and the lines belonging to another
element.
Superposition of absorbances from molecules
Diffusion of incoming light on small solid particles formed in
the atomizer during nebulization; the other metal salts present in a high
concentration in the sample becomes insoluble.
The detector, whatever it is, can only measure a light intensity. It is necessary to
so convert this signal to absorbance and, in addition, correct this absorbance from the
non-specific absorptions (NSA).
The different signals to be processed are as follows:
Incident intensity I0
Intensity emitted by the atomizer E
Emerging intensity I + E
To these three basic measures, essential for the determination of absorbance,
are added those allowing the correction of nonspecific absorptions (NSA):
Incident intensity of the corrector (it is also equal to I)0)

Emerging intensity after non-specific absorptions I‘

Correction of nonspecific absorptions
The role of the correctors is to automatically measure the ANS due to the
interferents of all kinds in order to subtract them from the total absorbance. For the
for the correction to be valid, it needs to be made as close as possible to the
characteristic wavelength 0λ
a) Correction by Deuterium lamp:

11
 Atomic Absorption Spectroscopy

The deuterium D2 source is a continuous source. There is an alternation of the 2.

sources (primary source of the hollow cathode lamp (HCL) and the source at D2).
When the device is equipped with a deuterium corrector, a lamp at
deuterium, producing a continuous light throughout the ultraviolet, is placed of
such that the optical paths of the lights coming from the two sources are
identical. The atomizer (flame or oven) is alternately illuminated by the two
radiations.
The monochromator selects the characteristic line of the analyte λ0 while it
let the light from the continuous source pass across the entire width of the strip
passerby.
We can assume that the luminous intensity at each point of the bandwidth
is the same.
The two integrated incident intensities are equal. One is represented by the
width of the line, the other is distributed over the bandwidth
When it is the primary source that illuminates the atomizer, the light is attenuated by
specific and non-specific absorptions. And when the deuterium lamp
illuminate the atomizer the light is attenuated by non-specific absorbances.
AS = Atotale - Ans
The use of the deuterium corrector involves the superposition of the two.
incident beams.
The deuterium corrector can only correct the ANS between 190 and 390 nm and the
Correction is only valid if the ANS are below a certain level.

b) The Smith-Hieftje effect correction

The operation of the corrector is based on a specific power supply of the
hollow cathode lamp in two phases. During the first phase, the lamp is
normally powered (low current) and the emission line is normal; then,
for a brief moment, it is overloaded (high current). In this case, the
the density of the atomic cloud at the end of the cathode increases and the atoms
can absorb the photons that come out of the cathode. We also observe a
broadening of the emission line due to the increase in current.
The center of the emission line is thus absorbed and it appears as
doubled. This phenomenon is known as strip reversal.

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 Atomic Absorption Spectroscopy

When the hollow cathode is normally powered, absorbance is measured.

total.
When the lamp is overdriven, if the line reversal is sufficient, the two
components will be evenly distributed on either side of the absorption line
atomic. We then only measure the background. By subtraction, we obtain
the specific absorbance.
Corrected = Total (normal line) - Specific (inverted lines).

c) The correction by Zeeman effect:

It is the phenomenon of the decomposition of spectral lines that we observe.
when a light source is placed in a magnetic field. The lines
absorption spectrum are divided into multiplets with symmetrical distribution by
regarding the frequency of the line for which the magnetic field is zero.

The Zeeman effect can be demonstrated from a light source placed

in the air gap of a magnet or an electromagnet with polar parts that are
breakthroughs along their axis to allow observation parallel to the lines of
magnetic field strength (which should be in the range of a few thousand)
Gauss).
a) If the field is null (H = 0), we observe the line of wavelength λ.
b) If the observation is made in the direction of the magnetic field (axis of the pieces
polar), the primordial radiation λ has disappeared and has been replaced by a doublet whose
The components are circularly polarized, one to the left, the other to the right.
c) If the observation is made in the direction perpendicular to that of the field
magnetic, we observe the presence of a triplet whose three components are
linearly polarized:
the central component, called π, is located at the wavelength λ; it
transport the vibrations parallel to the direction of the field;

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 Atomic Absorption Spectroscopy

The symmetric components called Ω and Ω' are found at the wavelengths
λ-∆λetλ+∆λ; they carry vibrations perpendicular to the direction of
champ.
The sum of the light intensities π and Ω is equal to the intensity of the line.
primary (for which H = O).
This is generally in this direction of observation that the Zeeman effect is applied.
profit for analysis by atomic absorption because of specific absorption (that due to
(to the dosed element) occurs only on π, while the unwanted absorptions
intervene both on Ω and Ω'.

The magnetic field can be applied either directly on the source of emission
(direct Zeeman effect), either on the atomization source (inverse Zeeman effect). The
spectrometry that uses the direct Zeeman effect employs a magnetic field
perpendicular to the emitted flux (transverse effect); for reasons that are both
theoretical and practical, it seems that the longitudinal Zeeman effect is not
valid in association with the direct Zeeman effect. The inverse Zeeman effect can, in
revenge, to be exploited with a transverse or longitudinal effect. Finally, one last
possibility that captures the authors' attention is the modulation of the field
magnetic at a given frequency.
Normal Zeeman effect: TRIPLET
Ba 553.6 nm
Cd 228.8 nm
Pb 283.3 nm 217.0 nm
Zn 213.8 nm
Anomalous Zeeman effect: MULTIPLET

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 Atomic Absorption Spectroscopy

As 193.7 nm
Bi 223 nm
With 324.8 nm
Mn 279.5 nm
Se 196, nm
Hg 435.8 nm
At 589.9 nm
2. Chemical Interferences
They are due to the formation in the flame of refractory oxides or of little salts.
volatiles. We also speak of matrix effect. They result from modifications in the
source of atomization, processes of dissociation, oxidation-reduction or
deionization. This is due to the alteration of the number or the rate of formation
of the analyte atoms, alteration resulting from the disturbance in the place
of atomization.
They are due to the formation of refractory oxides or low-volatile salts in the
flame.
Formation of oxides
Example: Al_2O_3, AlOH.Al2O3, AlOH.
To avoid this phenomenon, a so-called reducing flame is used:
Low oxygen content in the gas.
use nitrous oxide (N2O) as an oxidizer
Formation of refractory salt
Ca3(PO4)3 + 3LaCl2⟶La3(PO4)3 + 3CaCl2
To eliminate these refractory salts such as Ca_3(PO_4)_3, a salt (buffer) is added.
spectral) whose cation is capable of forming even more insoluble salts and
susceptible of giving more volatile salts with the element to be measured.
The most commonly used salts are:
Strontium chloride: SrCl_2
Lanthanum chloride: LaCl2

3. Physical Interferences
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 Atomic Absorption Spectroscopy

They are generally related to the physical properties of the studied solutions.
The type of interference is more noticeable in SAAF where the introduction of the sample
is ensured by a nebulization system; a change, for example, in the
Viscosity between the standards and the samples can introduce errors.
In SAAE, transport interferences will be limited only to the errors caused.
by the differences in viscosity between the standards and the samples, but in a
to a lesser extent than in SAAF.

To correct these interferences, we:

Make the solution iso-viscous: add albumin or wetting agents.
Use the method of calibration by standard additions (known additions).

4. Ionization Interferences:
Ionization interferences occur when the analyte is an element.
easily ionizable, because any atom that ionizes can no longer be measured. One chooses
so temperature conditions that prevent ionization. However,
It cannot always be avoided: the presence of another element more easily.
Ionisable modifies the ionization equilibrium of the analyte. It can be added deliberately.
to reduce the ionization of the analyte (buffer effect) and thus increase
absorbance.
Correction:
If we want to measure alkaline earth metals (e.g. Ca), to avoid ionization, we add
in the solution to dose elements that ionize more (e.g.: an alkali)
The Ca is protected. - To measure the alkalines, there is an element that ionizes more.
more easily than them: a tantalum salt. There is protection of the alkali, because this salt
supports ionization

V. APPLICATIONS:

The SAA allows the analysis of almost all metals and metalloids (Cu, Zn, Pb,
Cr, Fe, Cd, etc.…) in biological, metallurgical samples,
archaeological, pharmaceutical, and atmospheric. It therefore covers a vast
range of applications :
Zinc dosage in insulin or zinc oxide preparations.
Cobalt dosage in Vitamin B12.

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 Atomic Absorption Spectroscopy

Dosage of mercury in organo-mercurial antiseptics.

Measurement of Al and Mg in gastric dressings.
Dosage of Mg in nutritional supplements.
Dosage of Ca in Ca-based preparations.
The search for Cd, Zn in injectable preparations {plastic adjuvants}.
Search for impurities.
Analysis of drinks.
Dosage of trace elements and toxic residues in food.
Analysis of drinking water.
Analysis of plant and animal tissues, biological fluids.
Dosage of Ca, Sr, Zn in bones.

Sample preparation:
Atomic absorption spectrophotometry only requires two qualities of
samples: fluidity and homogeneity. Preliminary manipulations tend to
communicate, if necessary, these properties to the test samples
Case of liquid samples (blood, urine, cerebrospinal fluid,
punctuation fluids, aqueous humor,). The humors whose viscosity and
surface tension remains little different from that of water can be
nebulized directly: this is the case of urine for evaluation, for example.
from the copper industry, the value of which is usually known to be very low (lower than
at 0.1 mg/l.
A simple dilution may be necessary: dosing of magnesium in
the urine; in this case a possible precipitation of ammonium phosphate
Magnesian will be avoided by the addition of a few milliliters of hydrochloric acid.
for analysis
Plasma or serum blood have viscosity and content
organic levels too high to be used directly. a dilution of 1/5 otherwise.
on 1/10, it proved necessary to bring the viscosity back to an acceptable value,
close enough to that of the calibration solutions. This dilution leads to
a corresponding decrease in the concentration of the element to be measured,
desirable when the metal exists in significant amounts (calcium, magnesium,
for example), but undesirable in the case of trace element dosing. One can

17
 Atomic Absorption Spectroscopy

So resorting to three processes to adapt the physical characteristics and the

concentration of the test sampling to the requirements of the equipment used.
Mineralization:
This technique generally recommends a dry calcination in an oven at
5000 or 7000 C. This process is actually delicate to implement, because a
partial volatilization of the metal to be measured can occur (Mg and Zn for example) and
lead to default errors or, on the contrary, pollution from dust
The atmosphere will taint the result of error by excess.
Wet mineralization: A volume of reagent is used (some
acids, such as H2SO4, HNO3, H3PO4HCl, aqua regia (mixture of
HNO3and HCl in a ratio of 1:3), HF and/or oxidants, such as H2O2,
Hypochlorous acid4, H2SO4, HNO3in excess compared to the test dose, by heating to
a relatively low temperature (from 100 to 200°C) for quite some time
short (a few hours at most).
Dry mineralization: Heating is done at well-defined temperatures.
higher (around 450 to 550 °C)∘C, sometimes more), for a while
much longer, on the order of a day or more, using a low
amount of reagent, generally an oxidant (H2SO4HNO3, K2SO4, KNO3,
Mg(NO3)2, MgO ...) taken as a mineralization aid.
Deproteinization:
Calling upon the swift action of trichloroacetic acid to eliminate the
proteins whose presence causes interferences either of a chemical nature
(calcium), either of a physical nature (increase in viscosity).

Extraction :
An extraction step is necessary in some cases, such as for
the dosage in SAA-ET of total "non-food" or of interest arsenic
toxicological in the urine. Extraction in this case allows for the isolation of the forms
organic and inorganic substances of toxicological interest. It is a simple method and
rapid process that takes place in an acidic medium in the presence of potassium iodide to reduce the
arsenic species in corresponding forms of arsenic iodide complexes.
These are extracted by toluene and then taken up in aqueous media before
to be dosed in SAA-ET. Only inorganic arsenic and its methylated metabolites are
extracts, and dietary arsenic mainly in the forms of arsenobetaine or
d'arsenocholine is eliminated.

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 Atomic Absorption Spectroscopy

Case of organs: Metals often combine with

the organic substances making up the framework of animal tissue and lose,
this fact, their usual analytical traits. In the majority of cases, the
conventional techniques of inorganic chemistry cannot therefore
apply only to the product of the complete destruction of organic matter,
either by dry process (calcination) or by wet process, depending on the techniques
of toxicology.
In order to fully benefit from the advantages offered by the technique
of atomic absorption, it is important to seek simple procedures for implementation
quick execution solution, requiring only the use of volatile reagents. In
Indeed, to avoid an additional burden on fixed assets, it is desirable
that the extraction methods do not use mineral salts such as
nitrate, the acid sulfate or potassium permanganate used in the
sulfonitric mineralization techniques of Pouchet, Meillère, and Deniges. The
connective tissues, scleroproteins, are often difficult to solubilize,
it also appears necessary to finely divide the test sample so that the reagents
may reach the entire parenchyma. The fresh organs will therefore undergo a
careful grinding, but it often seems more convenient to use organs
dried, easy to spray.
Two simple acid extraction processes generally yield satisfactory results:
Digestive nitro-hydrochloric acid:
Crushed or powdered organs are placed in a 100 ml conical flask. We
Add 1 ml of nitric acid and 1 ml of concentrated hydrochloric acid and 6 ml of water.
We heat it up, then a simple centrifugation allows us to eliminate the debris.
tissues that settle and possibly the fats that float. The dosage
is then carried out on the aqueous phase, possibly diluted if the concentration
The body to be dosed appears to be higher than that of the usual calibration range.
In the case of elements existing only in trace amounts, it is possible to concentrate
by evaporation the previous solution. The nitro-chlorhydric digestion applies
to metals weakly bound to proteins (alkalis and alkaline-earths, by
example).

Digestion nitro-perchloric.
At the test sample, precisely weighed in a conical flask of 125 or 250 ml, one
Add 1.5 ml of nitric acid, 10 ml of distilled water, and 1.5 ml of perchloric acid.

19
 Atomic Absorption Spectroscopy

Let it macerate for a few hours. The mixture is brought to a boil while stirring.
constant. A part of the liqueur is centrifuged to the exact volume before
to proceed with the analysis or, possibly, a new dilution. For the
elements with very rich connections to the organic matrix (Cu and Fe by
for example), it seems preferable this technique
2. Measures :
Atomic absorption spectrometry is a comparative analytical method;
it involves calibration.
External calibration: The most common calibration is obtained by measuring
the absorbance of solutions with progressive and known concentrations of analyte.
The concentration of the unknown solution is then directly deduced in
reporting its absorbance value on a previously prepared calibration curve
established. This method, called direct calibration or external calibration, applies
in relatively simple environments, where the matrix is sufficiently constant. It
One must always check that the range of measurements is linear.
Calibration by dose additions: If the matrix is unknown or too variable in the
series of analyzed samples, we have the method of standard additions. It
consists of adding increasing and known quantities of the element in question
Qito the studied solution. The volume of these successive additions must be sufficient
small enough that we can neglect the variations in the volume of the solution
of the sample, On obtains of concentrations croissants C i ’.
We then measure the absorbance of the obtained solutions. By plotting the curve
Absorbance = f(concentration), we get a line whose intersection with the axis
The x-axis represents the concentration of the analyte in the solution without addition.
Currently, modern devices equipped with a sample dispenser
can perform these operations fully automatically.

VI. ADVANTAGES AND DISADVANTAGES:

The SAA is a simple, rapid, sensitive quantitative method.
Very selective and relative (therefore a calibration curve must be made).
It requires a small amount of sample.
Standard solutions are easy to prepare.

However, one can note a number of limitations:

necessity to use a characteristic source for each element to be dosed,

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 Atomic Absorption Spectroscopy

Destructive analysis technique,

limited field of application almost exclusively to metals (Cu, Zn,
Pb, Cr, Fe, Cd, etc.…
The existence sometimes of severe chemical interferences.
The non-qualitative aspect of the technique imposes the knowledge of the elements.
to dose in order to choose the appropriate source.
Does not allow simultaneous analysis of elements.
Need to have sufficiently low concentrations (to comply with the
linear domain of the Beer-Lambert law.
VII. CONCLUSION:
Atomic absorption spectrometry is a very widespread method, thanks to its
low price, and considering its versatility and ease of use,
considered the reference method for quantitative elementary analysis
she has given rise to many fields (Biochemistry, Toxicology,
Environment … ).
Due to the limitations of the SAA, the fact that it requires for each element to
measure a characteristic light source, the complexity of matrices, as well as the
lack of control over potential disturbances that may influence sensitivity and
the accuracy of the analysis; the SAA is completed or replaced by another
spectral technique that does not require a dedicated lamp for each element and
which allows simultaneous multi-element analysis without prior selection called
atomic emission spectroscopy (AES).

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VIII. BIBLIOGRAPHIC REFERENCES:

Anne LE BOUIL et AL. Chronic arsenic poisoning (online document)
CHU Angers, Annals of Analytical Toxicology vol. XIII, n° 3, 2001,
[Link]
Analytical Chemistry, Atomic Absorption Spectrophotometry (online)
05/06/19: [Link] absorption spectrophotometry — Chemistry
Analytical, (consulted on 18/11/22).
Dr Bouklif.S, Atomic Absorption Spectroscopy, postgraduate course
2019/2020.
Elhajji, Spectroscopic Techniques, Chapter 4: Spectroscopy
atomic absorption (online document)Invalid input, please provide a text for translation.
Fundamentals of Atomic Spectroscopy, (electronic document), 2016,
[Link]
▸Chemical analysis methods, Chapter 1; Absorption spectrometry
atomic (online)[Link] on
18/11/22
▸Maurice Pinta et AL. Atomic Absorption Spectrometry: Application to
Chemical analysis. Chapter 3: Sources of atomization, Volume I, Paris, 1971,
p 181, 192.
Maurice Pinta et al. Atomic Absorption Spectrometry: Application to
Chemical analysis. Chapter 10: Sample preparation, Volume II,
Paris, 1971, p 541-545.
M. DE KERSABIEC and M. L. RICHARD, Possibilities of exploitation
the Zeeman effect for the correction of nonspecific absorptions in
atomic absorption. Practical applications, Analysis v. 10, no 5, p 207 to
215, 1982 (online)Invalid input. Please provide text for translation.(consulted on
18/11/22)
Yves Pegon, Measurement of Aluminium in biological fluids by
flame-free atomic absorption, Elsevier Scientific Publishing Company
(online), 1978,Invalid request. Please provide text for translation., (consulted
on 11/20/22.

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