ATOMIC

ABSORPTION
SPECTROSCOP
Y
PRANAV KHANDEPARKAR
07
M.PHARM SEM 1 PQA
MPAT
 CONTENTS

• INTRODUCTION
• PRINCIPLE
• INSTRUMENTATION
• INTERFERENCES
• APPLICATION

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 INTRODUCTION

• It was introduced by alan walsh in the mid 1950’s.

• Atomic absorption spectroscopy (AAS) is defined as the
measurement and interpretation of the absorbed radiation in
atomic level. This method is widely used for the determination
of the elements.
• It is a method of elemental analysis.
• It is the most powerful instrumental technique for the
quantitative determination of trace metals.
.
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 PRINCIPLE
• The absorption of energy by ground state atoms in the gaseous
state forms the basis of atomic absorption spectroscopy.

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 There are mainly three steps involved in the AAS:
• Desolvation by evaporating the solvent present in the sample
and leaves the sample completely dry.
• Vaporising the evaporated solid sample into the gas or vapour.
• Voltalisation by breaking the vaporised sample into the free
atoms.

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 • AAS is used to determine the absorbed light which directly gives the
concentration of the element present in the sample. The absorption is
directly proportional to the concentration which is derived from the Beer–
Lambert’s law.

• Total amount of light absorbed

(at υ) = (πe2 /mc)Nf

• where e is the charge; m is the mass of the electron; c is the speed of the
light; N is the total number of atoms; f is the oscillator strength; Π, e and
m are constants hence the equation become
• Total amount of light absorbed = K × N × f

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 INSTRUMENTATION
• Radiation Source
• Chopper
• Atomiser
• Monochromator
• Detector
• Amplifier & Recorder

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 Radiation source
1.Hollow Cathode Lamp
• It is the most common source for atomic absorption
measurements
• This type of lamp consists of a tungsten anode and a cylindrical
cathode sealed in a glass tube filled with neon or argon at a
pressure of 1 to 5 torr.
• The cathode is constructed of the metal whose spectrum is
desired or serves to support a layer of that metal.
• The efficiency of the hollow-cathode lamp depends
on its geometry and the operating voltage.

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 2.Electrodeless Discharge Lamp
• It is difficult to make stable hollow
cathodes from certain elements,
particularly those that are volatile, such as
arsenic, germanium, or selenium.
• An alternative light source has been
developed in the electrodeless discharge
lamp (EDL).
• It 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.

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 Chopper
• A rotating wheel is interposed between the hollow cathode lamp and
flame.
• It is interposed to break the steady light from the lamp into an
intermittent and pulsating light.
• This gives pulsating current in the photocell.
• There is also steady current caused by light which is emitted by the
flame
• But only the pulsating current is amplified and recorded and thus
absorption of light will be measured without interference of light from
the flame itself.

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 Atomiser
• It consists of nebulizer and atomiser which is used to convert the test solution
to gaseous atoms.
• In order to achieve absorption of atoms, it becomes necessary to reduce the
sample to the atomic state.This is done by
(a) Flame Atomisers
(b) Non-flame Atomisers

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 FLAME
• Important regions of flame are
 Primary combustion zone
 Secondary combustion zone
 Interzonal region

• oxygen or nitrous oxide must be used as

the oxidant for more difficult to atomize
samples

• The burning velocities are important

because flames are stable only in certain
ranges of gas flow rates. If the gas flow
rate does not exceed the burning
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velocity, the flame propagates back into
 Flame atomisers
1.total consumption burner
• The total consumption type of burner consists of
three concentric tubes.
• The sample solution is carried by a fine capillary
tube directly into the flame.
• The fuel gas and the oxidant gas are carried along
separate tubes so that they only mix at the tip of
the burner.
Advatages:
Simple to manufacture
All sample reaches the flame
Free from explosion hazard
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 2.Pre-mix or laminar flow burner
In the premixed burner, a mixture of the sample and
premixed gases is allowed to enter from the base.
In the pre-mix type of burner, the aerosol is
produced in a vaporising chamber.
The resulting fine mist is mixed with the fuel gas and
the carrier (oxidant) gas, and the mixed gases then
flow to the burner head.
The burner is a long horizontal tube with a narrow slit
along its length.
In addition to a long light path, this type of burner
has the advantages of being quiet .
Disadvantages:
• with solutions made up in mixed solvents, the more
volatile solvents are evaporated preferentially;
• a potential explosion hazard
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 Non flame atomisers
1.Electrothermal atomiser
• The graphite tube furnace consists of a hollow graphite
cylinder.
• The graphite tube is surrounded by a metal jacket
through which water is circulated and which is separated
from the graphite tube by a gas space.
• An inert gas, usually argon, is circulated in the gas
space.
• The graphite cylinder is then heated by the passage of
an electric current to a temperature which is high
enough to evaporate the solvent from the solution.
• The absorption signals produced by this method may last
for several seconds and can be recorded on a chart
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recorder.
 2.Cold vapour atomisation
• This procedure is strictly confined to the determination
of mercury.
• which in the elemental state has an appreciable
vapour pressure at room temperature so that gaseous
atoms exist without the need for any special
treatment.
• As a method for determining mercury compounds the
procedure consists in the reduction of a mercury (II)
compound with either sodium borohydride or (more
usually) tin(II) chloride to form elemental mercury.
• Hg2++Sn2+ Hg + Sn(IV)
• The mercury vapour is flushed out of the reaction
vessel by bubbling argon through the solution, into the
absorption tube.
• The determination is completed by measuring the
17 absorbance at 253.7 nm.
 3.Hydride generation technique
• Elements such as arsenic, antimony, and selenium are
difficult to analyse by AAS because it is difficult to reduce
compounds of these elements to the gaseous atomic
state.
• Compounds of the above three elements may be
converted to their volatile hydrides by the use of sodium
borohydride as reducing agent.
• The hydride can then be dissociated into an atomic
vapour by the relatively moderate temperatures of an
argon-hydrogen flame.
• The reaction sequence, in the case or arsenic, may be
represented as follows:

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 Monochromator
• In atomic absorption measurements, the most common
monochromators are prisms and gratings.
• The function of a monochromator is to select a given absorbing line
from spectral lines emitted from the hollow cathode.

Detector
• For atomic absorption spectroscopy, the photomultiplier tube is most
suitable.
• It has good stability if used with a stable power supply. It works
satisfactorily and enables to compare intense in a satisfactory
manner.

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 PHOTOMULTIPLIER TUBE
• It consists of an evacuated envelope which contains a
photocathode, a series of electrodes called dynodes, and
an anode .
• The photocathode is fixed to the terminal of the power
supply.
• As soon as a photon strikes the photocathode, an electron
is dislodged and the photon is accelerated to dynode I,
resulting in the liberation of two or more electrons from
this dynode. Similarly, the pattern continues.
• Thus, the current multiplied at each dynode and the
resultant electron current is received by the anode to
produce an EMF across R₁ which goes to the external
amplifier and read-out system.

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 Amplifier
• The electric current from the photomultiplier detector is fed to
the amplifier which amplifies the electric current many times.
• Generally, "Lock-in" amplifiers are preferred which provide a
very narrow frequency band pass and help to achieve an
excellent signal-to-noise ratio.

Read-out Device
• In most of the atomic absorption measurements, chart recorders
are used as read-out devices.
• The displacement is directly proportional to the input voltage.
• In some atomic absorption measurements, digital read-out
devices are the used.

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 Single beam atomic absorption
spectrophotometer

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 Double beam atomic absorption
spectrophotometer

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 Interferences
• They are mainly of 3 types
Spectral interferences
Chemical interferences
Ionisation interferences

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 Spectral Interferences
• This type of interference normally takes place when the
absorption of an interfering species either overlaps or lies very
near to the analyte absorption, with the result that resolution by
the monochromator almost becomes impossible.
(a) Spectral interferences caused either by the combustion
products which show broad-band absorption or the particulate
products which scatter radiation. In fact, both these products
distinctly lower the power of the transmitted beam of light and
ultimately give rise to positive analytical errors.
Remedy
(a) When the source of the combustion or particulate products is
the fuel and oxidant mixture alone, then a blank is aspirated into
the flame and the necessary corrections are effected from the
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observed absorbances.
 (b)For instance : Manganese triplet (at 4031°, 4033° and 4035° A) :
potassium doublet (at 4044° and 4047° A) and the gallium line (at 4033°
A).
Remedy
• The overlapping of this nature may be eliminated either by prior
chemical separation or by selection other spectral lines.

(c) Sample Matrix: it occurs when the source of scattering originates

right in the sample matrix itself. In such a situation, it has been noticed
that the power of the transmitted beam-designated as P, is reduced by
the non-analyte components, whereas the incident beam power-
designated as Po, is not ; thereby resulting in a positive error in
absorbance and hence in concentration.
Example : Determination of Barium in alkaline-earth mixtures affords a
potential matrix interference due to absorption. It has been observed
that an intense and useful absorption line for barium atoms, occurring at
553.6 nm, lies in the centre of a broad absorption band for Ca (OH) 2, that
26 extends from 540 to 560 nm.
 Remedy :
(1) The effect due to sample matrix is quickly and effectively eliminated by
replacing nitrous oxide for air as the oxidant for the acetylene, whereby the higher
temperature completely decomposes the Ca (OH)2 and eliminates the absorption
band.
(2) If the source of interference is known, an excess of the interfering substance
may be added to the sample as well as the standards ; provided the ‘excess’ is
sufficient enough with respect to the concentration from the sample matrix, the
concentration of the latter will thus become insignificant. Such an added substance
is sometimes referred to as a radiation buffer.

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 Chemical interferences
• They are more common.
(i) Chemical Interferences due to Anion (PO4 3–) : Phosphate ions
have been found to interfere with determination of Mg and Ca by
AAS. The absorption due to Mg and Ca are appreciably weaker in the
presence of PO4 3– ions than in their absence. This is evidently on
account of the formation of fairly stable phosphates of Mg and Ca
which do not readily split-up into the respective atoms in the mantle
of a flame.
Remedy :
The addition of an excess of strontium (Sr), or lanthanum (La), or thorium
(Th) ion remarkably minimizes the interference of PO4 3– ion in the
determination of Mg, and Ca by replacing the analyte in the analyte in
the compound formed with the respective interfering species. In short,
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these ions do combine preferentially with PO4 3– ions.
 (ii) Chemical Interference due to Cations : In certain specific cases
cations also interfere in atomic absorption measurements, for instance :
Boron interferes with Mg and Ca ; whereas aluminium interferes with
alkaline earth elements.
• Protective Agents : These agents are found to inhibit the interferences
by virture of their ability to form relatively stable but volatile species
with the respective analyte. There are three reagents that are employed
commonly for this purpose, namely :
(a) Ethylenediamine tetra-acetic acid (EDTA).
(b) 8-Hydroxyquinoline,
(c) Ammonium salt of 1-pyrrolidinecarbodithioic acid (APDC). EDTA helps
to eliminate the interferences of Al3, Si4+, PO4 3– and SO4 2– in the
determination of Ca

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 Ionization interferences
• Ionization interference, arises in atomic absorption if the flame temperature is
too high. When this occurs, a number of the vaporized atoms become ionized
by the flame. The resulting ions absorb at a different wavelength than the
vaporized atoms; the new wavelength will not be selected by the
monochromator, and low values result.
• Any attempt to alleviate the ionization interference by lowering the flame
temperature might result in incomplete dissociation and more chemical
interferences.

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 • When potassium compound is added to the sample it is vaporized and
ionized
K K+

.
Remedy :The interference is usually minimized by the addition of a more
easily ionizable element. For example, ionization interference of calcium may
be corrected for by the addition of large quantities of sodium or potassium
salts to the solution. Since each of these elements possesses a lower
ionization potential than does calcium, their electrons will be more easily
removed and calcium will remain as the vaporized metal .

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 Applications
• Qualitative analysis- In AAS different hollow cathode lamp is
used for each element to be tested. It means element which is
used in the construction of lamp can be detected. eg. Na, K, Ca
etc
• Quantitative analysis-It is based on the determination of the
amount of radiation absorbed by the sample.
• Simultaneous multicomponent analysis-If multi element
source is available one can do simultaneous multicomponent
analysis. Mitchell described this system and detected 8
elements using multielement source and detection system.
• Determination of metallic elements in biological
materials- AAS, is becoming important tool for the
determination of trace metals in biological materials.
Decomposition of biological material is done by treating with
32 50% hydrogen peroxide, sulphuric acid or nitric acid.
 • Determination of metallic elements in food industry-
Copper, zinc, cadmium and nickel are most common toxic
elements of interest found in food. These metals are analyzed by
digesting food products either with 50% hydrogen peroxide or
sulphuric acid or nitric acid.
• Determination of calcium,magnesium, sodium and
potassium in blood serum-The determination of these
elements in blood serum is of vital importance in diagnosing
pathological conditions and is done by AAS.
• Determination of lead in petrol

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 Referances
• Skoog DA, Holler FJ, Crouch SR. Principles of Instrumental
Analysis. 7th ed. Boston: Cengage Learning; 2018.
• Chatwal GR, Anand SK. Instrumental Methods of Chemical
Analysis. 5th ed. Mumbai: Himalaya Publishing House; 2014.
• Christian GD. Analytical Chemistry. 6th ed. University of
Washington: John Wiley & Sons, Inc.; 2003.
• Vogel AI, Mendham J, Denney RC, Barnes JD, Thomas MJK.
Vogel's Textbook of Quantitative Chemical Analysis. 5th ed.
London: Longman Scientific & Technical; 1989..
• Kar A. Pharmaceutical Drug Analysis. 2nd ed. New Delhi: New
Age International Publishers; 2005.

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