ATOMIC ABSORPTION

SPECTROSCOPY
(AAS)

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INTRODUCTION
Development of current database for mineral nutrients and
toxicants in foods
Able to measure mineral composition and trace elements in food
rapidly, accurately and precisely.
Essential Nutrient

Toxic risk

Calcium

Lead

Phosphorus

Mercury

Sodium

Nickel

Potassium

Cadmium

Chlorine

Arsenic

Magnesium
Iron
Iodine
Zinc
Copper
Selenium
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Chromium
Manganese

INTRODUCTION
AAS (Atomic Absorption Spectroscopy)
An analytical method based on the absorption of UV or visible
radiation by free atoms in the gaseous state
Atomic absoprtion spectra are produced when ground state atoms
(or ions) absorb energy from a radiation source, moving into an
excited state

AES (Atomic Emission Spectroscopy)

Measures emission of radiation from atoms excited by heat or
other means
Atomic emission spectra are produced when excited atoms emits
energy on returning to the ground state
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INTRODUCTION
Principles :
Atomic absorption uses the absorption of light to measure the
concentration of gas-phase atoms. Since samples are usually
liquids or solids, the analyte atoms or ions must be vaporized in a
flame or graphite furnace

Atomization process
All elements in foods are virtually present as compounds or
complexes and therefore must be atomized before atomic absorption
/ emission requirements can be made
Atomic absorption requires that atoms of the element of interest to be
in the atomic state (not combined with other elements in compound)
and well separated
Atomization involves separating particles into individual molecules
(vaporizaton) and breaking molecules into atoms. Accomplished by
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exposing the analyte to high temp. in a flame (or plasma)

INTRODUCTION
Atomization process (contd)
Solution containing the analyte is introduced into the flame as a fine
mist. The solvent quickly evaporates, leaving solid particles of the
analyte that vaporize and decompose to atoms that may absorb
radiation (atomic absorption) or become excited and subsequently
emit radiation (atomic emission)

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A schematic representation of the

atomization of an element in a flame.
The large circle at the bottom represents a
tiny droplet of a solution containing the
element (M) as part of a compound.

INSTRUMENTATION
1. Light / Radiation source
2. Atomizer (Nebulizer burner system or electrothermal furnace)
3. Monochromator (UV-Vis grating)
4. Detector (photomultiplier tube)
5. Readout device (analog or digital)

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Schematic of Flame AAS

INSTRUMENTATION

Sample in a solution form (usually aqueous solution)

nebulized
(dispersed into a tiny droplets)

mixed with fuel and oxidant

burned in a flame
(produced by oxidation of fuel by oxidant)

atoms and ions formed

(analyte compounds decomposed by the high temperature)

atomized sample quantity measured

(determine the attenuation of a radiation beam passing through the flame)
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INSTRUMENTATION
Light source
The light source is usually a hollow-cathode lamp of the element that
is being measured.
Lasers are also used in research instruments
The disadvantage of these narrow-band light sources is that only one
element is measureable at a time.
Hollow cathode lamp :
- Consist of a hollow tube filled with argon or neon (inert gas, low
pressure), an anode made of tungsten and a cathode made of the
metallic form of the element being measured.

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INSTRUMENTATION
Light source
-When voltage is applied across the electrodes, the lamps emits
radiation characteristic of the metal in the cathode. If the cathode
made of iron, an iron spectrum is emitted. When this radiation passes
through a flame containing the sample, iron atoms in the flame will
absorb some of it because it contains radiation of exactly the right
energy for exciting iron atoms.
-Use different lamp for each element analyzed.
Radiation reaching the monochromator comes from two sources :
a) The attenuated beam from the hollow cathode lamp, and
b) Excited atoms in the flame

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INSTRUMENTATION
Light source
Overcome problem : Instruments are designed to discriminate
between these two sources either :
a) by modulating the lamp so that the output fluctuates at a constant
frequency
b) by positioning a chopper perpendicular to the light path between
the source and the flame

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INSTRUMENTATION
Atomizer
AA spectroscopy requires that the analyte atoms be in the gas phase.
ions or atoms in a sample must undergo desolvation and vaporization
in a high temp. source such as a flame or graphite furnace
Source of energy for
atomization

Approx. Atomization
temperature (oC)

Analytical Method

Flame atomization

1700 3150

AAS, AES

Electrothermal (graphite
furnace) atomization

1200 3000

AAS (graphite furnace)

Inductively coupled
argon plasma

6000 - 8000

ICP-AES

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INSTRUMENTATION
Atomizer
Chopper a disk with segments removed. It is rotated at a constant
speed so that the light beam reaching the flame is either on or off at
regular intervals.
The radiation from the flame is continuous. Therefore, the radiation
reaching the detector consists of the sum of an alternating and a
direct signal.
Type of atomizers used in AAS :
a) Flame
b) Electrothermal
c) Cold vapour technique for mercury
d) Hydride generation
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INSTRUMENTATION
a) Flame Atomizer
Consist of a nebulizer and a burner

Schematic representation of a nebulizer-burner assembly for an AS.

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INSTRUMENTATION
a) Flame Atomizer
Nebulizer :
- Designed to convert the sample solution into a fine mist or
aerosol
- Accomplished by aspirating sample through a capillary into a
chamber where oxidant and fuel are flowing
- Chamber contains baffles which remove large-droplets, leaving a
very fine mist that is carried into the flame by the oxidant-fuel
mixture. Large droplets fall to the bottom of the mixing chamber
and are collected as waste
- Burner head contains a long, narrow slot that produces a flame
(5 10 cm in length). A long path-length increases the sensitivity
of the measurement.
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INSTRUMENTATION
a) Flame Atomizer
The flame serves as the sample compartment
Need to choose a flame temperature that will maximize atomization
and minimize ionization
The importance of flame temp. :
- Affect efficiency of converting compounds to atoms and ions
- Influences the distribution between atoms and ions in the flame.
Flame characteristics manipulated by :
i) Choice of oxidant and fuel
ii) Adjustment of oxidant-fuel ratio
Most common oxidant-fuel combination :
i) Air-acetylene
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ii) Nitrous oxide-acetylene

INSTRUMENTATION
a) Flame Atomizer
Types of flame :
i) Stoichiometric produced from stoichiometric amounts of
oxidant and fuel so that the fuel is completely burned and
oxidant is completely consumed. Yellow fringes.
ii) Oxiding produced by fuel-lean mixture. The hottest flame, has
a clear blue appearance.
iii) Reducing produced from a fuel-rich mixture. Relatively cool
flame, has a yellow colour

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INSTRUMENTATION
a) Flame Atomizer
Advantages :
i) Stable and easy to use but sensitivity is relatively low because
much of the sample never reaches the flame and the residence
time of the sample in the flame is short
ii) Uses slot type burner to increase the path length, and therefore
to increase the total absorbance (refer Beer-Lambert Law)
iii) Sample solutions are usually aspirated with the gas flow into a
nebulizing / mixing chamber to form small droplets before
entering the flame
iv) Flame AA can only analyze solutions, while graphite furnace AA
can accept solutions, slurries or solid samples.

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INSTRUMENTATION
b) Electrothermal Atomizer
Has several advantages over a flame
Much more efficient atomizer than a flame and it can directly accept
very small absolute quantities of sample
Also provides a reducing envt. for easily oxidized elements.
- Samples are placed directly in the graphite furnace and the
furnace is electrically heated in several steps to dry the sample,
ash organic matter, and vaporize the analyte atoms
Typically cylindrical graphite tubes connected to an electrical power
supply

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INSTRUMENTATION
b) Electrothermal Atomizer
Sample introduced into the tube through a small hole using microliter
syringe (sample volume : 5 10 L). During operation, system is
flushed with an inert gas to prevent the tube from burning and to
exclude air from the sample compartment. Tube is heated electrically.
Temperature increase causing sample solvent evaporation then
ashed. Temp. rapidly increases to 2000 3000oC will rapidly vaporize
and atomize the sample.

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INSTRUMENTATION
b) Electrothermal Atomizer
Advantage :
a) can accommodate smaller sample that are required for the flame
AS
b) detections lower limit
Disadvantage :
a) added expense of electrothermal furnace (more expensive)
b) lower sample throughput
c) more difficult operation
d) lower precision

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INSTRUMENTATION
c) Cold Vapour Technique (only for mercury)
Mercury is the only element that can exist as free atoms in the gaseous
state at room temp.
Mercury compounds in sample reduced to mercury element by action
of strong reducing agent, then carried into a stream of air / argon into
an absorption cell. Atomic absorption measured the same way as flame
and electrothermal instruments.
Advantage : very high sensitivity because all mercury in the sample can
be transferred to absorption cell and measured.

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INSTRUMENTATION
d) Hydride Generation Technique
Elements of volatile hydrides (e.g. As, Pb, Sn, Bi, Sb, Te, Ge and Se)
formed by reacting samples with sodium borohydride; then carried into
absorption cell and heated to decompose them into free atoms.
Advantage : sensitivity high because very little sample loss
Disadvantage : limited to realtively few elements that are cpable of
forming volatile hydrides.

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INSTRUMENTATION
Monochromator
Positioned in the optical path between the flame / furnace and the
detector
Purpose : to isolate the resonance line of interest from the rest of the
radiation coming from the flame / furnace and the lamp, so that only
radiation of desired wavelength reaches the detector

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INSTRUMENTATION
Detector / Readout
Detector : a photomultiplier tube (PMT) that converts the radiant
energy reaching it into an electrical signal
Instrument electronic subtract the direct signal and send only the
alternating signal to the readout. Eliminates contribution of emission
from elements in the flame to the final signal.
Signal is processed to produce an analog or digital readout.

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INTRODUCTION
Practical Considerations
a) Reagent
Mineral elements concentration in foods are at trace level, thus it
is essential to use highly pure chemical reagent and water for
preparation of samples and standard solutions.
Water purified by distillation, deionization, or combination of the
two.
Only reagent grade chemicals should be used.
Reagent blanks must always be carried through the analysis.

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INTRODUCTION
Practical Considerations
b) Standards
Quantitative AS depends on the comparison of the sample
measurement with appropriate standards.
Standard solution contain the analyte metal in known
concentration in a solution that closely approximates to the sample
solution in composition and physical properties.
A series of standards varying concentration should be run to
generate calibration curve.
Standards have to be prepared with extreme care. Accuracy to
analyte determination depends on the accuracy of the standard.
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INTRODUCTION
Practical Considerations
c) Labware
Vessels used for the sample preparation and storage must be
clean and free of element of interest.
Preferably use plastic containers. Glass containers has great
tendency to adsorb metal ions.

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