Successfully reported this slideshow.
You’ve unlocked unlimited downloads on SlideShare!
Interferences and control measures
High sensitivity techniques
Comparison of AAS with GFAAS and ICP-MS.
The first AAS was presented by Walsh and co-workers in
Melbourne in 1954, was a double beam atomic absorption
Walsh worked with Perkin-Elmer,
the first AAS instrument developed
by the company was MODEL 303.
Atomic Absorption Spectroscopy (AAS) is a quantitative
method of analysis that is applicable to many metals and
a few non-metals.
Almost every metallic element can be determined
quantitatively by using the spectral absorption
characteristics of atoms.
It is a very common technique for detecting and
measuring concentration of metals in the samples.
It can analyze over 62 elements.
THE DETECTABLE ELEMENTS ARE THOSE
WHICH ARE COLORED IN PINK
“When a beam of monochromatic radiation is passed
through the atoms of an element, the rate of decrease of
intensity of radiation is proportional to the intensity of
incident radiation as well as the concentration of the
This technique basically uses the principle that free
atoms (gas) generated in an atomizer can absorb
radiation at specific frequency.
The atoms absorb UV or visible light and make
transitions to higher electronic levels. AAS quantifies the
absorption of ground state atoms in the gaseous state.
The analyte concentration is calculated from the amount
of absorption . 6
The basic requirements are:
(1) a light source;
(2) a sample cell; and
(3) a means of specific light measurement.
The two most common line sources used in atomic
absorption are :
1. ‘‘Hollow Cathode Lamp(HCL)’’ and
2. ‘‘Electrodeless Discharge Lamp(EDL).’’
HOLLOW CATHODE LAMP(HCL)
Its an excellent, bright line source for most of the
elements determinable by atomic absorption.
Each HCL will have a particular current for optimum
In general, higher currents will produce brighter
emission and less baseline noise. As the current
continues to increase, lamp life may shorten and spectral
line broadening may occur, resulting in a reduction in
sensitivity and linear working range.
CATHODE AND ANODE
The anode and cathode are sealed in a glass cylinder
normally filled with either neon or argon at low pressure.
At the end of the glass cylinder is a window transparent
to the emitted radiation.
The cathode of the lamp frequently is a hollowed-out
cylinder of the metal whose spectrum is to be produced.
Its constructed from a highly pure metal resulting in a
very pure emission spectrum.
The ‘‘multi-element’’ lamp may provide superior
performance for a single element or, with some
combinations, may be used as a source for all of the
elements contained in the cathode alloy.
Anode is made up of tungesten. 11
DISADVANTAGES OF HCL:
A finite lifetime – due to depletion of the analyte element
from the cathode
Adsorption of fill gas atoms onto the inner surfaces of
the lamp – the primary cause for lamp failure
Some cathode materials can slowly evolve hydrogen
when heated – a background continuum emission
contaminates the purity of the line spectrum of the
element, resulting in a reduction of atomic absorption
sensitivity and poor calibration linearity.
ELECTRODELESS DISCHARGE LAMP(EDL)
A small amount of the metal or salt of the element for which
the source is to be used is sealed inside a quartz bulb.
This bulb is placed inside a small, self-contained RF generator
or ‘‘driver’’. When power is applied to the driver, an RF field
The coupled energy will vaporize and excite the atoms inside
the bulb, causing them to emit their characteristic spectrum.
They are typically much more intense and, in some cases,
more sensitive than comparable hollow cathode lamps. Hence,
better precision and lower detection limits where an analysis
is intensity limited.
EDL are available for a wide variety of elements, including
Sb, As, Bi, Cd, Cs, Ge, Pb, Hg, P, K, Rb, Se, Te, Th, Sn and
The portion of an atomic absorption spectrometer’s
optical system which conveys the light from the source
to the monochromator is referred to as the photometer.
Types: single and double beam photometers
SINGLE BEAM PHOTOMETER:
It is called ‘‘single-beam’’ because all measurements are based
on the varying intensity of a single beam of light in a single
optical path .
The primary advantage is – it has fewer components and is
less complicated than alternative designs.
Hence, its easier to construct and less expensive.
With a single light path and a minimum number of optical
components, single-beam systems typically provide very high
The primary limitation is – it provides no means to
compensate for instrumental variations during an analysis,
such as changes in source intensity. The resulting signal
variability can limit the performance capabilities of a single-beam
DOUBLE BEAM PHOTOMETERS
It uses additional optics to divide the light from the lamp
into a ‘‘sample beam’’ (directed through the sample cell)
and a ‘‘reference beam’’ (directed around the sample
The reference beam serves as a monitor of lamp intensity
and the response characteristics of common electronic
Therefore, the observed absorbance, determined from a
ratio of sample beam and reference beam readings, is
more free of effects due to drifting lamp intensities and
other electronic anomalies which similarly affect both
sample and reference beams. 17
SINGLE BEAM PHOTOMETER
DOUBLE BEAM PHOTOMETER
OPTICS AND THE MONOCHROMATOR SYSTEM
An important factor in determining the baseline noise in an
atomic absorption instrument is the amount of light energy
reaching the photomultiplier (PMT). Lamp intensity is
optimized to be as bright as possible while avoiding line
Light from the source must be focused on the sample cell and
directed to the monochromator, where the wavelengths of
light are dispersed and the analytical line of interest is focused
onto the detector.
Light from the source enters the monochromator at the
entrance slit and is directed to the grating where dispersion
takes place. The diverging wavelengths of light are directed
toward the exit slit. By adjusting the angle of the grating, a
selected emission line from the source can be allowed to pass
through the exit slit and fall onto the detector.
PRE – MIX BURNER SYSTEM
In ‘‘premix’’ design, sample solution is aspirated through
a nebulizer and sprayed as a fine aerosol into the mixing
Here the sample aerosol is mixed with fuel and oxidant
gases and carried to the burner head, where combustion
and sample atomization occur.
Fuel gas is introduced into the mixing chamber through
the fuel inlet, and oxidant enters through the nebulizer
Mixing of the fuel and oxidant in the burner chamber
eliminates the need to have combustible fuel/oxidant in
the gas lines, a potential safety hazard. 21
Only a portion of the sample solution is introduced into
the burner chamber by the nebulizer.
The finest droplets of sample aerosol are carried with the
combustion gases to the burner head, where atomization
The excess sample is removed from the premix chamber
through a drain which uses a liquid trap to prevent
combustion gases from escaping through the drain line.
The inside of the burner chamber is coated with a
wettable inert plastic material to provide free drainage of
excess sample and prevent burner chamber ‘‘memory.’’
A free draining burner chamber rapidly reaches
equilibrium, usually requiring less than two seconds for
the absorbance to respond fully to sample changes. 22
Impact devices are used to reduce droplet size further
and to cause remaining larger droplets to be deflected
from the gas stream and removed from the burner
through the drain.
Classified as: impact beads and flow spoilers.
IMPACT BEAD FLOW SPOILER
Spherical bead made of glass, silica or ceramic.
Normally used to improve nebulization efficiency.
Positioned directly in the nebulizer spray as it exits the
Normally do not improve nebulization efficiency.
The primary use is to remove the remaining large
droplets from the sample aerosol.
They are used in atomic absorption burner systems
normally are placed between the nebulizer and the burner
They typically have three or more large vanes
constructed from or coated with a corrosion resistant
Smaller droplets are transported through the open areas
between the vanes while larger droplets contact the vanes
and are removed from the aerosol.
Classified as :
HIGH SENSITIVITY NEBULIZER STEEL NEBULIZER
BURNER HEADS AND MOUNTING SYSTEMS
Burner heads typically are constructed of stainless steel
or titanium. All-titanium heads are preferred as they
provide extreme resistance to heat and corrosion
A ten-centimeter single-slot burner head is recommended
for air-acetylene flames.
A special five-centimeter burner head with a narrower
slot is required when a nitrous oxide-acetylene flame is
to be used.
A ‘‘quick change’’ mount may reduce or eliminate
entirely the need for realignment of the atomizer when it
is replaced in the sample compartment.
The intensity of the light is fairly low, so a
photomultiplier tube (PMT) is used to boost the signal
A detector (a special type of transducer) is used to
generate voltage from the impingement of electrons
generated by the photomultiplier tube
Precision in Atomic Absorption Measurements:
Precision will improve with the period of
time over which each sample is read. Where analyte
concentrations are not approaching detection limits,
integration times of 1 to 3 sec will usually provide
acceptable precision. When approaching instrument
detection limits where repeatability is poor, precision can
be improved by using even longer integration times, up
to 10 sec.
CALIBRATION OF THE SPECTROMETER:
In the linear region, data on as little as one standard and a
blank may be sufficient for defining the relationship
between concentration and absorbance. However,
additional standards are usually used to verify calibration
Where the relationship becomes nonlinear, more
standards are required for which the accuracy of a
calibration depends on the number of standards and the
equations used for calibration.
By comparing the light intensity that
has passed through the sample with
that of the same light after it has
passed through a blank, the
absorbance is measured.
The absorbance of different standard
solutions of a compound of the
element are also measured and a
calibration curve is constructed.
Absorbance is plotted against
concentration. We then use the
calibration curve to determine the
add an excess
will form a
Exhaust System: AAS flames produce large amounts of heat
& the resultant fumes & vapours may be toxic.
Gas Cylinders: should be located outside of the laboratory in
a cool well-ventilated area.
Flammable Solvents: The combination of flame & solvent is
a hazardous situation. Always use a solvent with the highest
flashpoint consistent with the analysis being conducted. Use
covered containers & the smallest practical volume.
Burners: Keep burners clear & do not allow them to block.
UV Radiation: Hazardous UV radiation is emitted by flames,
hollow cathode lamps, analytical furnaces. Never look directly
at any of these. Operate the AAS with the door or flame shield
closed and wear appropriate safety glasses.
LIMITATIONS OF FLAME AAS
Determinations of analyte concentrations in the mg/L
concentration region are routine for most elements.
The absorbance depends on the number of atoms in the optical
path of the spectrometer at a given instant.
The nebulization process draws sample into the burner
chamber at approximately 3-8 ml/min limiting the sample
introduction rate, and, therefore, the amount of sample
available for transport to the flame.
Only a small fraction of the sample nebulized ever reaches the
flame, with the remainder being directed to the drain.
Sample which is introduced into the flame is resident in the
light path for only a fleeting moment as it is propelled
upwards through the flame.
HIGH SENSITIVITY TECHNIQUES:
All these limitations in the flame AAS lead to the invent
of high sensitivity techniques.
1. The cold vapor mercury technique
2. Hydride generation technique
3. Graphite furnace atomic absorption
THE COLD VAPOR MERCURY TECHNIQUE
Hg is chemically reduced to the free atomic state by
reacting the sample with a strong reducing agent like
SnCl2 or NaBH4 in a closed reaction system.
The volatile free Hg is then driven from the reaction
flask by bubbling air or Ar through the solution.
Hg atoms are carried in the gas stream through tubing
connected to an absorption cell, which is placed in the
light path of the AAS.
Sometimes the cell is heated slightly to avoid water
condensation but otherwise the cell is completely
Improved sensitivity is
achieved through a 100%
sampling efficiency (which
can be further increased by
using very large sample
All of the mercury in the
sample solution placed in
the reaction flask is
chemically atomized and
transported to the sample
cell for measurement.
The detection limit for Hg
approximately 0.02 mg/L
Its limited to Hg, since no
other element offers the
possibility of chemical
reduction to a volatile free
atomic state at room
The theoretical limit to this
technique would be that
imposed by background or
contamination levels of
mercury in the reagents or
system hardware. 42
HYDRIDE GENERATION TECHNIQUE:
Samples are reacted in an external system with a
reducing agent, usually NaBH4.
Gaseous reaction products(volatile hydrides) are then
carried to a sampling cell in the light path of the AA
To dissociate the hydride gas into free atoms, the sample
cell must be heated.
The cell is either heated by an air-acetylene flame or by
For As, Bi ,Ge ,Pb ,Sb ,Se
,Sn ,Te the detection limits
well below the mg/L range
The extremely low detection
limits result from a much
higher sampling efficiency.
Separation of the analyte
element from the matrix by
hydride generation is used
to eliminate matrix-related
Its limited to As, Bi ,Ge, Pb,
Sb, Se, Sn, Te elements only.
Results depend on the
valence state of the analyte,
reaction time, gas pressures,
acid concentration, and cell
The formation of the analyte
hydrides is suppressed by a
number of common matrix
components, leaving the
technique subject to
GRAPHITE FURNACE ATOMIC ABSORPTION:
In this technique, a tube of graphite is located in the
sample compartment of the AAS, with the light path
passing through it.
A small volume of sample solution is quantitatively
placed into the tube, normally through a sample injection
hole located in the center of the tube wall.
The tube is heated through a programmed temperature
sequence until finally the analyte present in the sample is
dissociated into atoms and atomic absorption occurs.
Detection limits fall in the ng/L range for most elements.
The sample is atomized in a very short period of time,
concentrating the available atoms in the heated cell and
resulting in the observed increased sensitivity.
This uses only micro litre sample volumes, which is
compensated by long atom residence time in the light
Much more automated than the other techniques.
It can determine most elements measurable by AA in a
wide variety of matrices. 46
GRAPHITE FURNACE TECHNIQUE
It’s a non flame technique
1. The atomizer
2. The power supply and
3. The programmer
• Electrical contacts
Enclosed water cooled housing
• Inert purge gas controls
The atomizer is located in the sampling compartment of
the atomic absorption spectrometer, where sample
atomization and light absorption occur. The power
supply controls power and gas flows to the atomizer
under the direction of the programmer, which is usually
built into the power supply or spectrometer
A GRAPHITE TUBE FOR A TRANSVERSELY-HEATED
The power supply and programmer perform the
1. Electrical power control
2. Temperature program control
3. Gas flow control
4. Spectrometer function control
STEPS MAKE UP THE TYPICAL GRAPHITE
3) Cool down (optional)
5) Clean out
6) Cool down
1) Temperature: final temperature during step
2) Ramp time: time for temperature increase
3) Hold time: time for maintaining final
4) Internal gas: gas type and flow rate 51
Parameters AAS GFAAS ICP – MS
Temperature 2300°C – 2700°C 3000°C 6000°C
Radiation used UV ,VISIBLE UV, VISIBLE ----
Detection limit Ppm Ppb Ppt
68+ 50+ 82
Sample throughput 10-15 sec per element 3-4 min per element All elements <1
Large Very small Very small to medium
Isotopic analysis No No Yes
items and utilities
hollow cathode lamps
hollow cathode lamps
graphite tubes and
Shriver and Atkins’ Inorganic Chemistry, Fifth Edition
Concepts, Instrumentation and Techniques in Atomic Absorption
Spectrophotometry by Richard D. Beaty and Jack D. Kerber
An elementary overview of elemental analysis by thermo elemental