Introduction

 Absorption

spectrophotometer is used to measure
concentration by detecting absorption of electromagnetic
radiation by atom rather than by molecules.
 It is a very common technique for detecting metals and
metalloids in sample.
 Widely used in clinical laboratories to measure elements
such as aluminum, calcium, copper, lead, lithium,
magnesium, zinc, and other metals
 Introduced in 1955 by Alan Walsh in Australia
 First commercial atomic absorption spectrometer was
introduced in 1959
 Principle

 Atomic absorption is an absorption spectrophotometric
technique in which a metallic atom in the sample absorbs
light of a specific wavelength.
 The element is not appreciably excited in the flame, but is
merely dissociated from its chemical bonds (atomized) and
placed in an unexcited or ground state (neutral atom).
 This ground state atom absorbs radiation at a very narrow
bandwidth corresponding to its own line spectrum
• A hollow cathode lamp with the cathode made of
the material to be analysed is used to produce a
wavelength of light specific for the atom.
• Thus, if the cathode were made of sodium, sodium
light at predominantly 589 nm would be emitted by
the lamp.
• When the light from the hollow cathode lamp enters
the flame, some of it is absorbed by the ground-
state atoms in the flame, resulting in a net decrease
in the intensity of the beam from the lamp.
• This process is referred to as atomic absorption
• Concentration measurements are usually
determined from a working curve after calibrating
the instrument with standards of known
concentration.
 Instrumentation

1. Light source
2. Chopper
3. Atomizer
4. Monochromators
5. Detector
7. Read out device
 Light source in AAS


Cathode Lamp
• Cathode--- in the form of a cylinder, made of the element being studied in the
flame
• Tungsten Anode
• Filled with an inert gas (neon or argon) sealed in a glass tube
• Quartz or special glass that allows transmission of the proper wavelength, is
used as a window
• Applying a potential difference(300-500V) between the
anode and the cathode leads to the ionization of some
gas atoms.
• These gaseous ions bombard the cathode and eject
metal atoms from the cathode in a process called
sputtering.
• Some sputtered atoms are in excited states and emit
radiation characteristic of the metal as they fall back to
the ground state.

Electrodeless Discharge Lamp

• Consists of an evacuated tube in which the metal of
interest is placed.
• The tube is filled with argon at low pressure and sealed
off.
 Chopper

• A rotating wheel is interposed between the hollow cathode
lamp and flame .
• It is interposed to break the steady light coming from the
lamp into pulsating light which is used to measure the
intensity of light absorbed by elements without
interference by radiation from the flame itself.
• Pulsating light gives pulsating current in photocell.
• There is also steady current caused by light which is
emitted by flame.
• But only pulsating current is amplified and recorded
 Atomizer

• Atomization is separation of particles into
individual molecules and breaking molecules into
atoms .
• This is done by exposing the analyte to high
temperatures in a flame or graphite furnace
• Atomiser converts the liquid into small droplets
which are easily vaporised.
• Types of Atomisers :-
1.Flame atomizer:-
2.Non-flame atomizer (Electrothermal atomizer)
 Flame Atomizer

 Nebulization • Conversion of the liquid sample to a
fine spray
 Desolvation • Solid atoms are mixed with the
gaseous fuel.
 Volatilization • Solid atoms are converted to a
vapour in the flame.
 There are three types of particles that exist in the
flame: 1) Atoms 2) Ions 3) Molecules
 Flame Atomizer

Advantages Limitations
 Short analysis time  Requires flammable gases
possible  Unattended operation is not
possible because of
 Good precision flammable gases
 Easy to use  Must not contain excessive
 Cheap amounts of dissolved solids
 Sensitivity
 Non- flame Atomizer

 aka Electro Thermal Atomizer
 The graphite furnace is an electro thermal atomizer
system that can produce temperatures as high as
3,000°C.
 The heated graphite furnace provides the thermal energy
to break chemical bonds within the sample held in a
graphite tube, and produce free ground state atoms.
 The ground-state atoms are capable of absorbing energy,
in the form of light, and are elevated to an excited state.
 The amount of light energy absorbed increases as the
concentration of the selected element increases
 Graphite Furnace AAS
Atomizer

Advantages Limitations
 Small sample sizes ( as low  Very slow
as 0.5 uL)
 Very little or no sample
 Fewer elements can be
preparation is needed analyzed
 High sensitivity due to entire  Poorer precision
sample is atomized at one  More chemical
time
 free atoms remain in the
interferences
optical path longer  Method development
 Reduced sample volume requires skill
 Ultra trace analysis possible  Expensive consumables
(graphite tubes)
 Monochromator

 It is used to separate out all of the thousands of lines.
 Without a good monochromator, detection limits are
severely compromised.
 A monochromator is used to select the specific
wavelength of light which is absorbed by the sample, and
to exclude other wavelengths.
 The selection of the specific light allows the
determination of the selected element in the presence of
others.
 They are of two types: 1) Prism 2) Diffraction Grating
 Detector

The light selected by the monochromator is
directed onto a detector that is typically a
photomultiplier tube , whose function is to convert
the light signal into an electrical signal
proportional to the light intensity.
The processing of electrical signal is fulfilled by a
signal amplifier.
The signal could be displayed for readout , or
further fed into a data station for printout by the
requested format
 Read-out Device

 The output from the detector is suitably amplified and
displayed on a readout device like a meter or a digital
display.
 It is capable of displaying the absorption spectrum as
well absorbance at specific wavelength
 Nowadays the instruments have microprocessor
controlled electronics that provides outputs
compatible with the printers and computers
 Thereby minimizing the possibility of operator error in
transferring data.
 Applıcatıons

 Clinical analysis - Analyzing metals in biological fluids
such as blood and urine.
 Environmental analysis - Monitoring our environment – eg
finding out the levels of various elements in rivers,
seawater, drinking water, air, and petrol.
 Pharmaceuticals- In some pharmaceutical manufacturing
processes, minute quantities of a catalyst used in the
process (usually a metal) are sometimes present in the
final product. By using AAS the amount of catalyst
present can be determined.
 Industry- Many raw materials are examined and AAS is
widely used to check that the major elements are present
and that toxic impurities are lower than specified .Ex. in
concrete, where calcium is a major constituent, the lead
level should be low because it is toxic.
 Applications

 Mining- By using AAS the amount of metals such as
gold in rocks can be determined to see whether it is
worth mining the rocks to extract the gold .
 Trace elements in food analysis
 Trace element analysis of cosmetics
 Trace element analysis of hair