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AAS e-Book

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Lab-Training.com

AAS e-Book

By

Dr. Saurabh Arora

Founder : Lab-Training.com
Director : Auriga Research Ltd.
E-mail : saurabharora@arbropharma.com

Dr. Deepak Bhanot

Vice President : Training & Development
E-mail : deepak.bhanot@aurigaresearch.com

Auriga Research Ltd.

Division of
Arbro Pharmaceuticals Ltd.
Analytical Division,

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Authors Profile
Dr Saurabh Arora is a trained pharmacist with Masters and Doctorate degrees in
pharmaceutics from reputed Indian institutes NPIER and Jamia Hamdard University
respectively.
Managing Director
Auriga Research Ltd.
He has setup 2 contract laboratories and a clinical research company along with managing and
growing the existing business. His organization has grown multiple folds and he hasbeen
fortunate to spearhead the growth initiatives backed by a team of over 250 employees..
Specialties: Formulation Development, Analytical Development, Chromatography, Mass
spectroscopy, GMP. GLP, GCP, Laboratory designing, Residue analysis, Project management,
International business and all that goes into growing and managing a business
Founder
Lab-Training.Com
Lab-Training.Com is developing and offering a series of free and paid E-Learning courses on
various analytical and laboratory techniques. He is responsible for the course concepts, course
content creation and review and course execution.
Founder
Food Safety Helpline
Food Safety Helpline has been established to help Food Business Operators implement the Food

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Safety and Standards Act

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Dr Deepak Bhanot is a seasoned professional having nearly 30 years expertise beginning from
sales and product support of analytical instruments. After completing his graduation and post
graduation from Delhi University and IIT Delhi he went on to Loughborough University of
Technology, UK for doctorate research in analytical chemistry. His mission is to develop
training programs on analytical techniques and share his experiences with broad spectrum of
users ranging from professionals engaged in analytical development and research as well as

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young enthusiasts fresh from academics who wish to embark upon a career in analytical industry.

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Knowledge grows when shared with others. Our belief in this has contributed immensely
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Knowledge does not discriminate between national boundaries color of skin, religion,
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AAS e-Book

Table of Contents
Introduction to Atomic Absorption Spectroscopy course ............................................................................. 8
Scope of Spectroscopic Analysis ................................................................................................................ 10
Advantages of spectroscopic techniques............................................................................................. 10
Application areas of spectroscopic analysis........................................................................................ 11
Types of spectroscopic analysis .......................................................................................................... 11
Evolution of Atomic Absorption Spectroscopy .......................................................................................... 13
Introduction to AAS component parts ........................................................................................................ 16
Burner system........................................................................................................................................ 17
Monochromator .................................................................................................................................... 17
Detector .................................................................................................................................................. 17
Double Beam Schematic ....................................................................................................................... 18
Types of Light Sources in AAS .................................................................................................................. 19
Hollow Cathode Lamps ........................................................................................................................... 19
Multi element Hollow Cathode Lamps ................................................................................................... 20
Limitations of Hollow Cathode Lamps .................................................................................................... 20
Electrodeless Discharge Lamps ............................................................................................................... 21
Flame Atomic Absorption Spectroscopy .................................................................................................... 22
Nebuliser ................................................................................................................................................ 22
Spray Chamber ..................................................................................................................................... 23
Burner Head .......................................................................................................................................... 24
Graphite Furnace Atomic Absorption Spectroscopy .................................................................................. 25
Limitations of flame AAS ......................................................................................................................... 25
Benefits of graphite furnace analysis...................................................................................................... 25
Graphite furnace components ................................................................................................................ 26
Limitations of graphite furnace analysis ................................................................................................. 27
Dispersion and Resolution of Light in Atomic Absorption Spectroscopy .................................................. 28

Spectral Interferences ........................................................................................................................... 32

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Non-spectral interferences ................................................................................................................... 31

Interferences in Atomic Absorption Spectroscopy ..................................................................................... 31

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Background correction in Atomic Absorption Spectroscopy ..................................................................... 34
Background Correction ........................................................................................................................... 34
Zeeman Background Correction ............................................................................................................. 36

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10 Interview questions in Atomic Absorption Spectroscopy ..................................................................... 38

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Introduction to Atomic Absorption Spectroscopy

course
It is possible to fly without motors, but not without knowledge and skill
Wilbur Wright

Atomic Absorption Spectrometer

The overwhelming response to the free e-book HPLC and GC courses has encouraged us to
move ahead with the free AAS course. We understand that everyone has busy work schedules
and today's hectic life style leaves you little or no time to refer to voluminous books to learn any
new technique. However, for sustained growth learning has to be adopted as a lifelong habit.
In an effort to make your learning task easy we have embarked upon the e-book which comprises
of 10 Chapters. Each Chapter comprising of about 300 to 400 words will provide functional
aspects of AAS and also present useful practical tips. Reading a Chapter and understanding it
will not take more than about 10 min and you'll get ample time to assimilate the contents before

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you move to the next Chapter.

AAS e-Book
AAS has emerged as a major analytical technique in diverse fields such as environmental
monitoring, mining and geology, oceanographic studies, studies on agricultural crops and soils,
pharmaceuticals, foods and beverages, petroleum and petrochemicals, forensic investigations and
hydro geological investigations.
The free program is designed to give an insight into the technique and once your interest is
captivated you can opt for our more elaborate online certificate program which will be
announced in due course. It will provide you an opportunity to interact with various learners and

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experts across the globe.

AAS e-Book

Scope of Spectroscopic Analysis

"If you want to increase your success rate, double your failure rate"
-- Thomas Watson, Sr. founder of IBM

Spectroscopic analysis is based on an atom or compound's interaction with electromagnetic

radiation of specific wavelength. Spectroscopy provides information on chemical identity of a
compound, quantity present and structure based on the technique selected and the wavelength of
electromagnetic spectrum. Commonly used spectroscopic techniques in any laboratory are UV
VIS spectroscopy, FT IR spectroscopy, Atomic Absorption spectroscopy and ICP/ ICP MS
spectroscopy.

This topic will introduce you briefly to the different spectroscopic analysis techniques commonly
used in laboratories.

Advantages of spectroscopic techniques

Rapid analysis information is available in a matter of seconds as compared to minutes

or even hours in other conventional techniques

Nondestructive most spectroscopic methods are non-destructive in nature and there is

100%recovery of sample after analysis

Micro analysis generally the methods can be adapted to micro volume analysis when
quantity of sample is limited.

High sensitivity inherent sensitivity of spectroscopic techniques coupled with advances

in detection technology provide unparalleled sensitivity. Advancements in hyphenated
analytical techniques such as GC IR, TGA IR, GC-MS and LC MS have lowered
detection and identification to levels which were not imagined earlier.

spectroscopic techniques like FT IR and corrective action can be initiated without the
need of sample withdrawal and off-line analysis.

Spectroscopic detection has been adapted to a number of techniques such as HPLC

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Real-time monitoring manufacturing processes can be monitored real-time using certain

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AAS e-Book
Application areas of spectroscopic analysis

Scope of Spectroscopy

Application scope of spectroscopic analysis is virtually unlimited. Knowledge gained in

such analysis can contribute to

Understanding constitution of matter from atoms to complex molecules

Studies on diverse materials existing in nature from deep sea studies to space missions

Investigations of crime samples

Analysis and development of whole range of man-made materials of human consumption

Studies on environmental samples

Mineralogy

Types of spectroscopic analysis

UV VIS Spectroscopy
Radiation in the UV and visible region of electromagnetic radiation interacts with organic
molecules or atoms selectively to give information on presence of absorbing entities. Absorption
radiation results in shifts of electrons within the electron levels of atoms and molecules.

Radiation in the IR region results in changes in bonding in terms of vibration frequencies,

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rotation and vibration energies depending on the wavelength within the IR region. Such

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FT IR Spectroscopy

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information provides the basis for both qualitative and quantitative levels of IR absorbing
groups.
Atomic absorption Spectroscopy
Atomic absorption spectroscopy is based on absorption by ground state atoms of an element
present in the sample which is atomized in the flame or graphite furnace. Depending on
absorption of selected wavelength of the element the concentration is estimated. The technique
provides valuable information on concentration of required elements present in the sample.
Concentrations are possible in ppm or ppb levels depending on source of sample excitation.
ICP/ICP MS
ICP uses a plasma source of excitation of sample. The temperature of the plasma is 2 to 3 orders
of magnitude above the flame AAS methods. The technique affords sensitivity upto ppb or even
sub ppb levels. ICP MS technique further extends separation of ionized species based on
charge to mass ratio by a quadrupole mass selector. This facilitates analysis of a number of

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elements that trace levels simultaneously.

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Evolution of Atomic Absorption Spectroscopy

Trace metal studies on composition of materials has been the oldest branch of analytical
chemistry. Traditional gravimetric techniques still constitute the backbone of most undergraduate
educational laboratories but instrumental methods are fast replacing them due to advantages of
speed and precision. Atomic Absorption Spectroscopy to this day has maintained a top-notch in
most of the academic and industrial laboratories due to its affordability and range of applications.
Bunsen and Kirchoff studied the sodium spectrum and came to the conclusion that every element
has its own unique spectrum in the vapour phase implying that a metal in atomic state can absorb
radiation at same wavelength at which it emits it. This is the founding principle of atomic
absorption spectroscopy. In 1859 Kirchoff showed that the Fraunhofer lines in the suns
spectrum were atomic lines due to presence of various elements in the sun's atmosphere.
Spectrochemical analysis had its origins with the work of Bunsen and Kirchoff but found little
application until 1930s. Modern Atomic Absorption Spectroscopy began in 1955 by a team of
Australian scientists led by Alan Walsh at CSIRO (Commonwealth Science and Industry
Research Organization) division of chemical physics, Australia. Walsh suggested the use of
hollow cathode lamps to provide the appropriate wavelength and use of a flame to generate
neutral atoms that would absorb the incident radiation in proportion to the concentration present

Early Day Atomic Absorption Spectrometer

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in the traversed path.

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Early day instruments did not have much limitations but the technique had its own
inherent limitations such as

Flame is not an ideal atomizer because of partial atomization, loss of sensitivity due to
background interference and only a fraction of sample reaching the flame due to
nebulization and passes quickly through the light path

Sample has to be in liquid form and therefore solids will require pretreatment and
digestions

Only one element can be analyzed at a time

Modern Day Atomic Absorption Spectrometer

Modern developments and advances in electronics and automation did not eliminate such
limitations but made it possible to increase laboratory throughput through features such as:

Introduction of nitrous oxide acetylene flame by Willis in 1965. It extended the number
of elements which could be determined due to higher flame temperatures.

Introduction of techniques such as mercury hydride analysers afforded greater accuracy

High energy sources such as electrodeless discharge lamps for analysis of volatile
elements

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and precision for analysis of metals like Hg,Pb,Sn,As,etc.

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Multi element lamps for faster analysis of number of elements in a sample

Electrically heated graphite furnace analyzers for greater precision and handling of small
sample amounts for lower detection limits

Introduction of background correction techniques

Multi lamp holders to expedite warm up prior to analysis

Several manufacturers utilize the advanced features and provide their advantages in

Perkin Elmer

Agilent Technologies

Analytik Jena

Shimadzu

Aurora Biomed

Hitachi

GBC Scientific Equipment

BuckScientific

Thermo Scientific

Teledyne Leeman Labs

Skyray Instrument

PG Instruments

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competitive environment. Some of the reputed manufacturers are :

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Introduction to AAS component parts

"A day spent without learning something is a day wasted"
-- Anonymous
Atomic absorption occurs when a ground state atom absorbs light of a specific wavelength. The
amount of light absorbed is governed by Beer Lambert's law and will increase as the number of
atoms of the element in the light path increases. The component parts of Atomic Absorption
Spectrometer are similar to a UV -Vis spectrophotometer as both operate on same principle with
a basic difference that the sample cell of UV-Vis spectrophotometer is replaced by an
atomization source (flame or graphite furnace)

AAS Schematic Diagram

Light sources Hollow Cathode Lamps
The light source commonly used is a hollow cathode lamp. A different element hollow cathode
lamp is required for each element determination. Cathode is made of same metal that is to be

Lamps are made of glass with quartz windows and filled with an inert gas such as argon.

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Single element lamps are used commonly though multi-element lamps are also available.

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estimated in the sample.

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Light sources Electrodeless Discharge Lamps
Used for volatile elements such as As,Sb, Sn,Cd,Pb, etc
EDLs have greater lamp life and high energy throughput
Burner system
The burner assembly comprises of nebulizer to reduce the liquid sample to a fine aerosol, a spray
chamber and a burner head which is used to generate a flame to produce atoms of the same
elements that are present in the sample.
Monochromator
A monochromator disperses the incident light beam and permits the selected wavelength to reach
the detector.
Detector
Detector commonly used is a photomultiplier tube which produces a signal proportional to the
amount of light received by it.
In this section you shall be introduced to two working configurations, namely single beam and
double beam instruments

Single Beam Schematic Diagram

specific wavelength isolated by monochromator is led to the detector and the electrical signal
generated is proportional to the elemental concentration in the sample.

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This helps isolate and remove sample cell emissions from light emitted by the source. The

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Light from the source is modulated electronically or chopped mechanically by rotating chopper.

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Double Beam Schematic

Double Beam AAS Schematic Diagram

Light beam from source is split into two teams by the chopper. One beam passes directly
through the flame and the other beam passes round the flame. Detector response represents the
ratio of sample and reference beams. Fluctuations in light intensity are eliminated electronically
to get greater reliability of results.
The sensitivity is lower than single beam instruments but the popular acceptance of double beam

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configuration is due to advantage of elimination of background changes in the atomizer.

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Types of Light Sources in AAS

"A man is but the product of his thought, what he thinks, he becomes"
-- M.K. Gandhi
Light sources are generally of two types. You'd be familiar with continuum light sources such
as Sun or a light bulb which emit electromagnetic radiation in the wavelength range from about
250 to 700 nm in the visible region which we see as normal white light. The white light
comprises of several different wavelengths which constitute the colours of the rainbow. The
other type of light sources are line sources which emit light of a specific wavelength and it is
such light sources which are used in Atomic Absorption Spectroscopy. Now you shall be
introduced to such light sources
Hollow Cathode Lamps
A hollow cathode lamp gives a high intensity, narrow line wavelength of element to be
determined

The hollow cathode lamp consists of a glass cylinder filled with an inert gas usually Argon or

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Hollow Cathode Lamp

Neon at low pressure. The cathode is made from metal which is to be determined.. The emission

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Hollow Cathode Lamp Schematic

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line of the lamp corresponds with the absorption wavelength of the analyte. The end window of
the lamp is usually made of Quartz or Pyrex that transmits the spectral lines of the element to be
determined.
Following stages are involved in light emission from Hollow cathode lamp:

Sputtering filled gas is ionized when potential difference is applied between the anode
and the cathode. Positively charged inert gas ions strike the negatively charged cathode
and dislodge metal atoms.

Excitation sputtered metal atoms are excited to impact with the ionized gas

Emission light of wavelength specific to the element comprising the cathode is emitted
when the atom decays from the excited state to the normal state

Hollow cathode lamps have a shelf life as well as usage lifetime defined in milliampere hours.
Increasing current increases lamp intensity but excessive current reduces lamp life and also
results in self absorption broadening ,i.e, atoms in the hollow cathode lamp begin to absorb light
emitted from the hollow cathode lamp itself. This leads to lower absorbance and reduction in the
linear range of calibration curve.
Multi element Hollow Cathode Lamps
The cathode of mult ielement lamps is made from alloying compatible elements without
overlapping line spectra. Examples of such lengths are Ca-Mg,Cu-Fe-Ni, Cu-Fe-Mn-Zn, etc. All
elements of multi element hollow cathode lamps can be determined sequentially without need for
change of lamps in between. Multi element lamps provide advantages of cost, speed of analysis
but the sensitivity is lower in comparison to individual element determination by single element
lamp

Hollow cathode lamps have a shelf life

With the exception of multi element lamps the lamp needs to be changed for
determination of different elements

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Limitations of Hollow Cathode Lamps

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Sputtering deposits metal atoms on sides and end windows which affects lamp life and
more so for volatile elements

Some cathode materials liberate hydrogen on heating which contributes to continuum

background emission

Electrodeless Discharge Lamps

For most elements hollow cathode lamp is a satisfactory light source. In case of volatile elements
reduced lamp life and low intensity can be overcome by use of high energy throughput
electrodeless discharge lamps. Electrodeless discharge lamps are commonly available for Sb, As,
Bi, Cd, Cs, Pb, Hg, K, Rb, Sn, Te, etc.

Electrodeless Discharge Lamp Schematic

Electrodeless Discharge Lamp

An EDL consists of a quartz bulb filled with an inert gas containing the element or a salt of the
element for which the lamp is to be used. The bulb is placed inside a ceramic cylinder on which
antenna for a RF generator is coiled. When an RF field is applied to the bulb, the inert gas is
ionised and the coupled energy excites the vaporized atoms inside the bulb and causes emission
of characteristic light. EDLs offer advantage of lower detection limits. The useful life of an

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EDL is considerably longer than that of a hollow cathode lamp of same element.

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Flame Atomic Absorption Spectroscopy

"Formal education will make you a living, self education will make you a fortune"
-- Jin Rohn
Sample atomisation produces ground state atoms that are necessary for atomic absorption to take
place. This involves application of thermal energy to break the bonds that hold the atoms
together.
The complete atomisation assembly comprises of:

Nebuliser

Spray chamber

Burner Head

Each of the components of the atomisation assembly are discussed in detail below:
Nebuliser

AAS Nebulizer Schematic

AAS Nebulizer

Nebuliser converts the liquid sample into a fine spray or aerosol. In order to provide efficient
nebulisation for different sample solutions (aqueous or organic, acids or bases, etc) the nebuliser
should be adjustable and corrosion resistant. Stainless steel is commonly used but for corrosive

nebulisation efficiency for lowest detection limits.

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used. High sensitivity in combination with inert ceramic bead can be used to enhance

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solutions other corrosion resistant materials such as inert plastic, Pt/Ir or Pt/Rh alloy are also

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Spray Chamber

Spray Chamber

Aerosol from the nebuliser is led to the mixing or spray chamber. In this chamber the aerosol is
mixed with fuel and oxidant gases and carried to the burner head. Only a fraction of the sample
introduced by the nebuliser is used for analysis. An impact device prevents larger droplets from
reaching the burner as these would delay sample vaporisation and atomisation through short
transit through the flame. Only fine sized droplets are carried to the burner head

An impact device such as a flow spoiler or an impact bead is aligned at the exit of the aerosol
stream of the nebulizer. A flow spoiler is more efficient at removing large droplets whereas the
impact bead removes fewer large droplets and exhibits better sensitivity since more sample is led
to the burner. However, the increased number of large droplets may have undesirable effects and
increase interference.

Glass and ceramic impact beads can cause memory and contamination problems compared to the
chemically inert flow spoiler and for this reason flow spoiler is preferred for routine work and for
greater sensitivity impact bead is preferable. The excess sample is removed from the pre-mix
chamber through a drain. The drain uses a liquid trap to prevent combustion gases from escaping
through the drain line. The inside of the spray chamber is coated with wettable plastic material to

sample changes.

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burner chamber rapidly reaches equilibrium typically in less than two seconds for response to

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provide free drainage of excess sample and prevent burner chamber memory. A freely draining

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Burner Head

Burner Head

Burner heads are constructed of titanium to provide extreme resistance to heat and corrosion. A
10 cm single slot burner is recommended for air- acetylene flames. Its long length provides best
sensitivity. A special 5 cm burner head is recommended for nitrous oxide acetylene flame
applications. The flame can be rotated to provide reduced sensitivity.

Single slot 5 cm air-acetylene burner head is available when reduced sensitivity is required. It
can be rotated to provide further sensitivity redaction and it has a wide slot to prevent clogging

A 3- slot burner head is designed for analysis of samples having high concentration of dissolved
solids.

Majority of elements can be an analysed using air acetylene flames which have high
temperature range of 2150 C 2300 C. Nitrous oxide acetylene flames attain temperatures of
2600C- 2800 C and can be used for analysing refractory elements which form stable oxides at

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lower temperatures.

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Graphite Furnace Atomic Absorption Spectroscopy

"The only source of knowledge is experience"-- Albert Einstein
Flame atomic absorption spectroscopy is a well established and precise method for elemental
analysis giving concentration results in mg/L (ppm) levels. However, better sensitivity is
achievable using electro- thermal atomisation with a graphite furnace.
Limitations of flame AAS

Burner nebuliser is a rather inefficient sampling device. Majority of the sample gets
drained and the small fraction reaching the flame has a short residence in the light path

High sample consumption of the order of 3-5ml/min

Matrix interferences limit applications particularly in analysis of biological and

geological samples

Analysis limited to ppm concentration ranges

Benefits of graphite furnace analysis

Entire sample is atomised and the atoms are retained in the atomisation graphite tube for
extended user controlled time periods

Microlitre quantities of sample are sufficient and the quantity can be increased to 50
100 l to enhance sensitivity

Temperature programming steps help remove the solvent and major matrix interferences

Detection limits typically 100 - 1000 times better than flame techniques are achievable

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thereby giving routinely analysis in g/l(ppb levels)

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Graphite furnace components

Graphite Tube with Lvov Platform

Graphite tube serves as a sample cell as well as a heating element

Electrical contact cylinders provide electrical cconnection
onnection to the tube.Current flow provides
heating of tube and sample
Water cooling housing serves to cool the assembly
Inert gas protects heated tube from atmospheric oxidation. External gas stream surrounds the
outside of the tube and internal gas fflow
low purges the tube. Flow is reduced or completely stopped
during atomisation to increase sample residence time and improve signal output.
Quartz windows at each end of the tube help to seal the tube and allow light to pass through

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Power supply programmer controls current supplied to tube as covered by user program

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Graphite Furnace Components

Transverse heating provides uniform heating of graphite tube across its length. In end to end
heating there can be temperature gradient along the tube length. Lvov platforms delay the
vaporization and atomisation of the sample until furnace atmosphere has reached equilibrium
conditions.
Stabilised temperature platform furnace (STPF) was pioneered in 1970s by Perkin Elmer. It is a
combination of graphite tube quality, design and operational parameters to improve atomisation
and detection. Tube lifetime improvement is provided by using high-quality graphite for the
tubes., Platforms maximise power heating to virtually eliminate interferences and internal gas
stop increases sensitivity.
Limitations of graphite furnace analysis

Longer analysis time in comparison to flame analysis

Lesser number of elements analysed by furnace technique around 40 as compared to

about 70 in flame technique

Higher cost of graphite furnace assembly but it is also available as a switching option
with flame operation in most commercial instruments
Higher and more complex background levels require expensive background correction

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options

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Dispersion and Resolution of Light in Atomic

Absorption Spectroscopy
"Learn something new. Try something different. Convince yourself that you have no
limits"
-- Brian Tracy
Monochromator
A monochromator is a device that isolates and transmits a band of wavelength from a wider
range of wavelengths available at the inlet slit. The dispersion of light can be obtained by means
of a prism or diffraction grating. The Czerny- Turner monochromator using a pair of concave
mirrors and a plane grating is most widely used in atomic absorption spectroscopy.

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Czerny Turner Monochromator

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Broadband beam reaches the entrance slit positioned at the focal length of the collimating mirror
and the parallel beam is diffracted by the plane grating and after reflection from the second
mirror is focused on the exit slit. As each color (wavelength) arrives at a separate point in the
exit plane, a series of images are focused on the exit slit. As the exit slit has a finite width, parts
of nearby images overlap. The light leaving the exit slit contains the image of entrance slit along
with images of nearby colors Rotation of dispersion grating causes the band of colors to move
relative to fixed exit slit so that the desired entrance slit image can be centered on the exit slit.
Thus the range of colors leaving the exit slit is a function of slit width. Slit size is variable,
though usually not continuously.
Mirrors
Mirrors used in the monochromator must be highly reflecting in the wavelength range of interest.
This can be achieved by polishing the front surface with aluminium, silver or gold. The metal
layer is covered with a protective coating that prevents the metal from tarnishing.
Grating
The dispersion of light takes place on the grating. Parallel beam striking the grating leaves the
grating at slightly different wavelengths. The angle of dispersion at the grating is controlled by
the density of lines on the grating, i.e. number of lines/mm. High dispersion is achieved by
increasing the line density. In order to isolate desired line from nearby lines narrower exit slit is
used. The use of a wider slit width allows more light thereby enhancing sensitivity but at the cost
of resolution.
Blaze angle governs the efficiency of the grating. The slope of the triangular groove in a ruled
grating is adjusted to enhance the brightness of a particular diffraction order. The further
removed a given wavelength of light is from the wavelength for which the grating is blazed the
greater will be the extent of light loss at that wavelength.

900 nm. With a grating blazed somewhere in the middle of this range significant energy fallof

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occurs at the wavelength extremities due to the energy inefficiencies of the diffraction process.

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The wavelength range normally used in atomic absorption spectroscopy is from 185nm to about

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Dual blazed gratings with a blaze in both UV and visible regions offer better energy efficiency
over the entire wavelength range.
Monochromator Parameters
Slit Width
Slit width is the width in millimetres of the entrance and exit slist of the monochromator. Narrow
slit width gives better resolution. In standard monochromator design both entrance and exit slits
have equal width. Wider the slit widths more wavelengths passes through the monochromator.
Research grade instruments have user controlled slit widths.
Monochromator Focal Length
Greater the focal length of collimating mirrors the larger their resolution. The resolving power of
a monochromator is governed by both focal length and slit width.
Dispersion
The dispersion of a monochromator is characterized as the width of band of wavelengths per unit
of slit width, i.e, nm of wavelengths per mm of slit width
Spectral Bandwidth
Spectral bandwidth is the width of the triangle at the points where the light has reached half the
maximum value defined as Full Width at Half Maximum (FWHM)
Stray Light

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Stray light is light other than selected wavelength reaching the detector.

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Interferences in Atomic Absorption Spectroscopy

Interference is a phenomena that leads to changes in intensity of the analyte signal in
spectroscopy. Interferences in atomic absorption spectroscopy fall into two basic categories,
namely, non-spectral and spectral.
Non-spectral interferences affect the formation of analyte items and spectral interferences result
in higher light absorption due to presence of absorbing species other than the analyte element.

Interference in Atomic Absorption Spectroscopy

Non-spectral interferences
Matrix interference
When a sample is more viscous or has different surface tension than the standard it can result in
differences in sample uptake rate due to changes in nebulization efficiency. Such interferences

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are minimized by matching as closely as possible the matrix composition of standard and sample

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Chemical interference
If a sample contains a species which forms a thermally stable compound with the analyte that is
not completely decomposed by the energy available in the flame then chemical interference
exists. Refractory elements such as Ti, W, Zr, Mo and Al may combine with oxygen to form
thermally stable oxides. Analysis of such elements can be carried out at higher flame
temperatures using nitrous oxide acetylene flame instead of air-acetylene to provide higher
dissociation energy. Alternately an excess of another element or compound can be added e.g. Ca
in presence of phosphate produces stable calcium phosphate which reduces absorption due to Ca
ion. If an excess of lanthanum is added it forms a thermally stable compound with phosphate and
calcium absorption is not affected.
Ionization interference
Ionization interference is more common in hot flames. The dissociation process does not stop at
formation of ground state atoms. Excess energy of the flame can lead to excitation of ground
state atoms to ionic state by loss of electrons thereby resulting in depletion of ground state atoms.
In cooler flames such interference is encountered with easily ionized elements such as alkali
metals and alkaline earths. Ionisation interference is eliminated by adding an excess of an
element which is easily ionized thereby creating a large number of free electrons in the flame and
suppressing ionization of the analyte. Salts of such elements as K, Rb and Cs are commonly used
as ionization suppressants.
Spectral Interferences
Spectral interferences are caused by presence of another atomic absorption line or a molecular
absorbance band close to the spectral line of element of interest. Most common spectral
interferences are due to molecular emissions from oxides of other elements in the sample.
The main cause of background absorption is presence of undissociated molecules of matrix that

molecular species in the flame which may scatter light over a wide wavelength region. When this

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type of non-specific adsorption overlaps the atomic absorption of the analyte, background

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have broad band absorption spectra and tiny solid particles, unvaporized solvent droplets or

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absorption occurs. The problem is overcome by measuring and subtracting the background

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absorption from the total measured absorption to determine the true atomic absorption.

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Background correction in Atomic Absorption

Spectroscopy
"Tell me and I'll forget : show me and I may remember; involve me and i'll understand"
-- Chinese Proverb
Before you move to background correction it is necessary to understand what is background
absorption. The main reason for background absorption is presence of undissociated molecules
of matrix that have broadband absorption spectra and tiny solid particles in the flame which may
scatter light over a wide wavelength region. When this type of non-specific adsorption overlaps
the atomic absorption wavelength of the analyte the ground state absorption is cut. The problem
is overcome by measuring and subtracting the background absorption from the total of measured
absorption to determine true atomic absorption component
Background Correction
A number of background correction approaches have been proposed but we shall limit our
discussion to two main approaches which have found widespread application in commercial
instruments
Continuuum Deuterium Source Background Correction
Continuum background correction measures and compensates for any background component

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present in atomic absorption measurements

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D2 Lamp Background Correction Schematic

The broadband contimuum source emits light over a broad spectrum of wavelengths instead of
specific line.Spectral interferences which are caused by atoms with absorption lines very close to
the analyte absorption line or by fine structure in the molecular absorption profile can result in
either positive or negative errors in the measurement of the concentration of the analyte.
Fortunately spectral interferences are rare in flame atomic absorption which is a distinct
advantage over emission techniques
A common inexpensive technique for background correction in flame Atomic Absorption
Spectroscopy is deuterium background correction. The correction is effective over the
wavelength range of 180nm -420nm. Background level becomes significant at lower wavelength
range.
The cathode lamp and the deuterium lamp are sequentially pulsed with a chopper or
electronically with delay of about 2ms. When hollow cathode lamp is on and deuterium lamp off
total absorbance (AA + BG) is measured. When the HCL is off and the deuterium lamp on the
continuum energy recorded is (BG). The atomic signal is automatically calculated by subtracting
background from total absorbance.

The D2 lamp has a finite lifetime and requires periodic replacement

The D2 lamp and HCL light may not view the same portion of atom cloud in the flame
due to time lag. This could become significant at high background levels.

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Limitations of Continuum D2 background correction :

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Proper alignment of both light sources is required for right background correction

Correction is limited to the specific wavelength range of the two lamps

Intensities of both lamps if not similar will result in errors

Zeeman Background Correction

Zeeman Background Correction Schematic

Zeeman Background Correction is used mainly in graphite furnace atomic absorption systems.
When an atom is placed in a magnetic field and its absorption of observed in polarised light, the
normal single line is split into three components -, and +displaced symmetrically about
the normal position

Magnetic Field Off

Magnetic Field on

Free atoms show Zeeman splitting in a magnetic field but molecules, liquid droplets or solid

component is linearly polarised parallel to the magnetic field while the components are

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circularly polarised perpendicular to the magnetic field. A polariser is positioned in the optical

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particles show no Zeeman splitting and so advantage can be taken of polarised light. The

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system to remove the components of the transmitted radiation. This affords background
measurement at the exact analyte wavelength when magnetic field is applied. Since the
background is measured at the analyte wavelength and not averaged as in D2 system structural
molecular background and spectral interferences are easily corrected.
In AC modulated Zeeman system the combined atomic and background absorption is measured
while magnetic field is off. When the magnetic field is on, the detector measures only the
background absorption as the component is not detected. The difference between the two is the
Zeeman corrected atomic absorption signal
The magnetic field may be applied to the graphite tube with either transversally( perpendicular)
or longitudinal ( parallel) to the optical axis. A major advantage of longitudinal Zeeman
background correction is that the polariser is not needed to eliminate the component, thereby
providing higher light throughput by eliminating polarizer absorption
Advantages of Zeeman background correction :

Corrects high levels of background

Corrects at exact analytical absorption line

Requires only a single standard light source. Alignment problems of multiple light
sources are not encountered

Limitations of Zeeman background correction :

More expensive than continuum background correction

Loss in sensitivity for some elements due to splitting of and components which may
overlap

So far you have been introduced to the basic concepts and principles of Atomic Absorption
Spectorscopy. The next Chapter is a set of questions which you may face at time of interview

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Spectrometer.

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when you apply for a suitable opening in a laboratory equipped with an Atomic Absorption

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10 Interview questions in Atomic Absorption

Spectroscopy
In this Chapter we have posed before you some typical questions which you could come across
during interview sessions when you apply for a job involving use of atomic absorption
spectrometers. We have deliberately not provided you with tailored made answers and the
objective is to motivate you to look for the answers. We are sure that if you review the earlier
Chapters on the topic you'll find all your answers. This exercise will encourage you to search for
solutions that you are faced with not only in atomic absorption spectroscopy but in other areas of
your activity as well.
Q1. Explain briefly the principle of operation of an Atomic Absorption Spectrometer?
Q2. What is the difference in operation principle of AAS and AES?
Q3. Why is nitrous oxide used as oxidant in some applications?
Q4. What are the advantages of using Electrodeless discharge lamps over Hollow cathode lamps
as light sources for analysing some elements?
Q5. What are the advantages of double beam over single beam operation?
Q6. What is the role of a flow spoiler or an impact bead in the spray chamber? Explain the
benefits of each over the other.
Q7. What is the role of a monochromator in the atomic absorption spectrometer?
Q8. What do you understand by deuterium background correction and what are its limitations?

,etc What alternate sample treatment option is commonly used for such analysis?

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Q10. What are the limitations in analysis of analysis of volatile elements such as As, Sn, Pb,Sb

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Q9. Explain the benefits of graphite furnace analysis over flame analysis?

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Conclusion
We believe that you enjoyed the free e- course on AAS. The course provided you an insight into
the components of AAS system and their individual contribution towards the overall accuracy
and precision of your results. Apart from a general introduction the course was designed keeping
in mind the requirements of the AAS user. Without going into mathematical treatment of the
subject an attempt has been made to convey the basics concepts and offer practical tips on
effective utilization of the AAS system.

In case the e- course has awakened your desire to go deeper into the subject you are welcome to
join the Certificate Course on AAS which shall be available round the year after its launch. For
more details on this advanced programme please await our announcement on the site.

Once again we take the opportunity to thank you for your interest. Please feel free to participate
actively by contributing articles in areas off your interest and offer your valuable comments and

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suggestions.

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