ATOMIC ABSORPTION

SPECTROSCOPY

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 introduction
• 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 of
the atoms.
• It is a very common technique for detecting and
measuring concentration of metals in the
samples.
• It can analyze over 62 elements. 2
 Atomic Spectroscopy

Absorption Spectroscopy: Emission Spectroscopy:

AAS FES, ICP-AES(OES)

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Atomic Spectroscopy

Ion Emission

Atom Emission

E – energy difference between two levels;

h – Plank’s constant, 6.626068 × 10-34 m2kg/s;
c – speed of light,
λ – wavelenght, nm 4
 ATOMIC ABSORPTION SPECTROSCOPY
• Atomic absorption spectroscopy is a technique for
determining the concentration of a particular metal
element in a sample. Atomic absorption
spectroscopy can be used to analyze the
concentration of over 62 different metals in a
solution.

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 Principle:
• “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
solution.”
• 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 .
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Atomic Absorption Spectroscopy:

A= ε b C

Bear-Lambert Law

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• The equation representing the Beer’s law:
A=εbc
• Where
– A is absorbance (no units, A = log10 P0 / P ).

– ε is the molar absorbtivity (is a measure of the

amount of light absorbed per unit concentration) with
units of L mol-1 cm-1.

– blength
is the path length of the sample that is, the path
of the cuvette in which the sample is contained.
We will express this measurement in centimeters.

– c is the concentration
-1
of the compound in solution,
expressed in mol L .

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 Principle of Atomic Absorption
Spectrophotometer
Atomic Absorption Spectroscopy, AAS Excited state E1

Absorption

e
Ground state E0

e
Excited state E1
Atomic Emission Spectroscopy, AES

Emission

e
Ground state E0

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 THEORY OF ATOMIC ABSORPTION
SPECTROSCOPY

• Absorption therefore is carried out by unexcited atoms.

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• The total amount of light absorbed may be given mathematically by the
following expressions:

Total number of light absorbed = πe2/mc Nf

Where,
e= is the charge on the electron of mass
m= mass of electron
c= is the speed of light
N= is the total number of atoms that can absorb at frequency in the light
path
v= frequency
f= is the oscillator strength or ability of each atom to absorb at frequency
π= is constant

The above equation can be written as:

Total amount of light absorbed= Constant x N x f

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 Elementary Theory
Characters of the atomic absorption spectrum

ΔE = E1 – E0 = hc / 

E1 - excited state
E0 – ground state
h – Planck’s constant
c – velocity of light
 - wavelength

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 Instrumentation:
• The basic requirements are:
(1) a light source( continous source,line source)
(2) a sample cell; and
(3) a means of specific light measurement.

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 1. RADIATION SOURCE

The main sources used for atomic absorption are:

I. Hollow Cathode Lamp (HCL)
II. Electrode less Discharge Lamp (EDL)

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 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 performance.
• 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.

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 Electron discharge lamp
• 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 is created.
• The coupled energy will vaporize and excite the atoms inside the bulb,
causing them to emit their characteristic spectrum.
• 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 Zn.

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 Sample Introduction Methods:
1- Introduction of solution samples:
a) Nebulizers
b) Electrothermal (furnace) evaporator
c) Hydride generation (As, Sb, Sn, Se, Bi, Pb )
d) Cold vapor atomization ( Hg )

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18
 Electrothermal evaporator :

There is no nebulziation, etc. The sample is introduced

as a drop (usually 10-50 uL)

• The furnace goes through several steps:

a- Drying (usually just above 110 deg. C.)
b- Ashing (up to 1000 deg. C)
c- Atomization (Up to 2000-3000 C)
d- Cleanout (up to 3500 C or so). Waste is blown out with a
blast of Ar.

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 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 spectrometer.
• 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 electricity
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A hydride generation and atomization
system

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 Advantages
• For As, Bi ,Ge ,Pb ,Sb ,Se
,Sn ,Te the detection limits
well below the mg/L
range are achievable.
• 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 interferences.
limitations:
 Its limited to As, Bi ,Ge, Pb,
Sb, Se, Sn, Te elements only.
 Results depend on the
valence state of the analyte,
reaction time,concentration,
and cell temperature.
 The formation of the analyte
hydrides is suppressed by a
number of common matrix
components, leaving the
technique subject to
chemical interference. 22
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.
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Advantages: limitations:
• Improved sensitivity is  Its limited to Hg, since no
achieved through a 100% other element offers the
sampling efficiency (which possibility of chemical
can be further increased
by using very large sample reduction to a volatile free
volumes) atomic state at room
• All of the mercury in the temperature.
sample solution placed in  The theoretical limit to this
the reaction flask is technique would be that
chemically atomized and
transported to the sample imposed by background or
cell for measurement. contamination levels of
• The detection limit for Hg mercury in the reagents or
approximately 0.02 mg/L system hardware. 24
2. ATOMIZERS

There are two types of atomizers which

are used in Atomic Absorption
Spectroscopy.
• Flame Atomizers
• Electrothermal Atomizers

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 SAMPLE NEBULIZER
ASSEMBLY
FLAME
(ATOMIZATION
OCCURS)

CONVERSION
INTO FINE MIST
AEROSOL
& SMALL
MIXES WITH
DROPLETS OF
COMBUSTION
SOLUTION
GASES
ASPIRATED
INTO SPRAY
CHAMBER
(MIXING
CHAMBER) 26
Flame atomization
Processes occurring during atomization

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Sample to be Nebulizer:
nebulized is
sample+fuel+
taken
(aspirated) oxidant
via a capillary fine mist or
tube aerosol

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Natural Gas Air 1700-1900 39-43

Natural Gas Oxygen 2700-2800 370-390

Hydrogen Air 2000-2100 300-440

Hydrogen Oxygen 2550-2700 900-1400

Acetylene Air 2100-2400 158-266

Acetylene Oxygen 3050-3150 1100-2480

Acetylene Nitrous Oxide 2600-2800 285

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30
 GFAAS ( ELECTROTHERMAL
DISCHARGE)
Graphite furnace atomic Graphite tube
absorption spectrometry (GFAAS)
(also known as Electro thermal Enclosed
Atomic Absorption spectrometry water cooled
(ETAAS)) is a type of spectrometry housing
that uses a graphite-coated furnace to
vaporize the sample. Transparent
windows
Instead of employing the high
temperature of a flame to bring about
the production of atoms from the Inert purge gas
sample and it is non-flame methods control
involving electrically heated graphite
tubes or rods. Electrical
contact
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Simple Schematic
Working

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 Working of GFAAS
•Aqueous samples should be acidified (typically with nitric acid, HNO3) to a
pH of 2.0 or less. Discoloration in a sample may indicate that metals are
present in the sample. For example, a greenish color may indicate a high
nickel content, or a bluish color may indicate a high copper content. A good
rule to follow is to analyze clear (relatively dilute) samples first, and then
analyze colored (relatively concentrated) samples. It may be necessary to
dilute highly colored samples before they are analyzed.
•After the instrument has warmed up and been calibrated, a small aliquot
(usually less than 100 microliters (µL) and typically 20 µL) is placed, either
manually or through an automated sampler, into the opening in the graphite
tube.
•The graphite furnace is an electrothermal 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 and produce free
ground-state atoms. Ground-state atoms then are capable of absorbing
energy, in the form of light, and are elevated to an excited state.
Contd....
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 Graphite furnace AAS
Sample holder: graphite tube
Samples are placed directly in the
graphite furnace which is then
electrically heated.
Beam of light passes through the tube
Three stages:
1. drying of sample
2. ashing of organic matter (to burn off
organic species that would
interfere with the elemental analysis.
3. vaporization of analyte atoms

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Advantages of GFAAS
Greater sensitivity and detection limits
(hundred- or thousand fold improvements in the
detection limit compared with flame AAS) than
other methods.
Direct analysis of some types of liquid
samples.
Some solid sample do not require prior
dissolution.
Low spectral interference.
Very small sample size (as low as 0.5µL).

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Disadvantages of GFAAS
expensive
low precision
low sample throughput
requires high level of
operator skill

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Applications of GFAAS
GFAA has been used primarily for analysis of low
concentrations of metals in samples of water. The
more sophisticated GFAAs have a number of lamps
and therefore are capable of simultaneous and
automatic determinations for more than one element.
for the quantification of beryllium in blood and
serum.

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 - Wavelength selectors
- Produces
monochromatic light

Consists of:
1) Entrance slit
2) Diffraction grating
3) Exit slit

Diffraction gratings are

mostly used rather than
prisms

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Grating and Prism

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 Detector
• The intensity of the light is fairly low, so a
photomultiplier tube (PMT) is used to boost the
signal intensity
• A detector (a special type of transducer) is used to
generate voltage from the impingement of
electrons generated by the photomultiplier tube

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• Atomic Absorption spectrophotometric measurements are
done extensively by using;
 Single-Beam AA Spectrophotometer
 Double-Beam AA Spectrophotometer

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SINGLE BEAM PHOTOMETER

DOUBLE BEAM PHOTOMETER

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 Sensitivity and detection limit
• Sensitivity
number of ppm of an element to give 1%
absorption.
• Limit of detection: dependent upon
signal:noise ratio.

• S/N is directly proportional to the

Light intensity reaching detector

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 INTERFERENCES &
CONTROL MEASURES

NON SPECTRAL SPECTRAL

Matrix Background
Chemical Ionization
Absorption

Method of add an excess

of another using a adding an Continuum
Standard Zeeman
element or hotter excess of Source
Background
Additions Background
compound flame. an element Correction
Correction
which which
will form a is very
thermally
stable easily
compound ionized
with the
interferent 44
 Spectral interference
This may be caused by direct overlap of the
analytical line with the absorption line of the
matrix element.
HOW TO OVERCOME ?
 By choosing an alternate analytical wavelength
 By removing the interfering element from the
sample.

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 Chemical interference
Formation of compound of low
volatility

Decrease in calcium absorbance is

observed with increasing
concentration of sulfate or
phosphate

 By increasing flame temperature

Use of releasing agents (La 3+ )
Cations react with the interferent releasing the analyte
Use of protective agents:
They form stable but volatile compounds with analyte.

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 Ionization interference
Ionization of ground state gaseous atom with in a flame will reduce extent of
absorption in AAS.
M ↔ M+ + e

HOW TO MINIMIZE:

Low temperature of the flame

Addition of an excess of ionization suppressant e.g. the alkali metals (K, Na,
Rb, and Cs)

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 Background correction
• Background interferences are usually caused by absorption by
the polyatomic species or scattering of radiation within the
cell.
• Background interferences is usually greater at shorter
wavelength.
• Correction :
Background interferences are corrected by using double beam
spectrophotometer, and continous radiative source.

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 Characters of the atomic absorption spectrum

Natural broadening
Determined by the lifetime of the excited state a（10-5 nm）

Doppler Broadening （10-3 nm）

Results from the rapid motion of atoms as they emit or
absorb radiation

Collisional Broadening
collisions between atoms and molecules in the gas phase
lead to deactivation of the excited state and thus
broadening the spectral lines 49
Flame Emission Spectroscopy (FES)

 Propane-butane flame ( 2000 – 3000 º C);

 Optical filter is used to monitor for the selected emission wavelength
produced by the analyte;
 Suitable for elements with low excitation energy (Na, K, Li, Rb and Ca).

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 Principle Process
Solution containing a metallic salt is aspirated
into a flame
Evaporating solvent leaving the solid,
Dissociating solid by vaporization into
gaseous atom
Raising atoms of the metal to higher
energy level by heat of the flame,
Emitting energy in the form of radiation.

• For e.g.:-
Orange color is imparted to the flame by
calcium compounds.
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Atomic Emission Spectrometer

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 Advantages of plasma:
1- more complete atomization
2- fewer chemical interferences
3- low ionization interference effects
4- atomization occurs in a chemically inert
environment
5- temperature cross section of plasma is
relatively uniform
6- wider linear range
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ICP-AES(EMISSION SPECTROSCOPY)
Basics
Atomic emission spectroscopy measures the intensity of light
emitted by atoms or ions of the elements of interest at specific
wavelengths;
Inductively Coupled Plasma spectrometers use emission
spectroscopy to detect and quantify elements in a sample;
 ICP-AES uses the argon plasma (6000-10000º C) for atomization
and excitation of the sample atoms;
 ICP-AES determines approximately all of the elements except
gases and some non-metals (C, N, F, O, H). 54
Schematic diagram of the processes in the ICP

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 ICP spectrometer

Main Systems

Sample Introduction System: to deliver the sample solution to the plasma.

Consists of pump, nebulizer and spray chamber.

 Plasma: to generate the signal. Plasma is forming in the torch from gas argon.

 Optics: to measure the signal.

Computer with appropriate software: for controlling the instrument and

measuring process.
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 ICP-AES: Plasma
Inductively Coupled Plasma Source
A plasma is a hot, partially ionized
gas. It contains relatively high
concentrations of ions and electrons.

Argon ions, once formed in a plasma, are

capable of absorbing sufficient power from
an external source to maintain the
temperature at a level at which further
ionization sustains the plasma indefinitely.
The plasma temperature is about 10 000 K.

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ICP-AES: Plasma
Inductively Coupled Plasma Source

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 ICP-AES: Radial (SOP) and axial (EOP)

SOP: Side-on-Plasma EOP: End-on-Plasma

 more suitable for hard matrices (concentrated samples);  more suitable for light matrices;
 alkali metals (Na, K, Li) calibration is more linear;  alkali metals (Na, K, Li) calibration is less linear;
 less spectral interferences;  more spectral interferences;
lower sensitivity (Limit-of-Detection is higher); higher sensitivity (Limit-of-Detection is lower);

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 Characteristic of the ICP:
• High temperature
• Long residence time
• few ionization interferences
• Free atoms formed in nearly chemically inert
environment
• Molecular species absent or present in very low
levels
• No electrodes
• No explosive gases

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 ICP –AES/OES
• Offer several advantages over
flame/electrothermal
Lower inter-element interference (higher temperatures)

With a single set of conditions signals for no. of elements can be recorded
simultaneously

Lower LOD for elements resistant to decomposition

Permit determination of non-metals (Cl, Br, I, S)

• Disadvantages:
• More complicated and expensive to run
• Require higher degree of operator skill

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 SPIKING
Spiking means deliberately adding analyte to an
unknown sample
Involves:
preparation of sample and measurement of absorbance

Addition of standard with known concentration,

measurement of absorbance

From difference in absorbance, calculate amount of

analyte
(use Beer’s law for calculations)
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 USES OF SPIKING
• Add spike at beginning of sample preparation

• Process sample with and without spike

• Difference should correspond to amount

spiked

• Deviation allows to calculate recovery factor

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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
Elements applicable 68+ 50+ 82
to
Sample throughput 10-15 sec per 3-4 min per element All elements <1
element minute
Sample volume Large Very small Very small to medium
required
Isotopic analysis No No Yes
typical consumable acetylene/nitrous argon gas argon gas
items and utilities oxide gases hollow cathode lamps
required (compressed air graphite tubes
source)
hollow cathode lamps

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 AAS Advantages and Disadvantages
Advantages
1. High selectivity and sensitivity
2. Fast and simple working
3. Doesn’t need metals separation
4. Specific because the atom of a particular element can
only absorb radiation of their own characteristic
wavelength

Disadvantages
1. Analysis doesn’t simultaneous
2. Can’t used for elements that give rise to oxides in
flames
3. Limit types of cathode lamp (expensive)
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 APPLICATION

• A) Quantitative analysis
• B) Qualitative analysis
• C) Simultaneous multicomponent analysis
• D) Determination of metallic element in
biological materials
• E) Determination of metallic element in food
industry
• F) Determination of Ca, Mg, Na, K in blood
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