INTRODUCTION

• Atomic Absorption Spectroscopy is a very common technique for detecting

metals and metalloids in samples.
• It is very reliable and simple to use.
• It also measures the concentration of metals in the sample.
• Atomic Absorption Spectroscopy is an analytical technique that measures
the concentration of an element by measuring the amount of light that is
absorbed at a characteristic wavelength when it passes through cloud of
atoms
• As the number of atoms in the light path increases, the amount of light
absorbed increases.

HISTORY OF ATOMICABSORPTION SPECTROSCOPY

• The Atomic Absorption Spectroscopy was first used as analytical technique

in the second half of 19th century by Robert Bunsen and Robert Kirchhoff.
• The modern form of Atomic Absorption Spectroscopy was largely
developed during the 1950s by a team of Australian chemists.
• They were led by Sir Alan Walsh at the Common wealth Scientific and
Industrial Research Organization (CSIRO)

PRINCIPLE OF ATOMICABSORPTION SPECTROSCOPY

• The technique uses basically the principle that free atoms generated in an
atomizer can absorb radiation at specific frequency.
• Atomic Absorption Spectroscopy quantifies the absorption of ground state
atoms in the gaseous state.
• The atoms absorb UV or visible light & make transition to higher electronic
energy level. The analyte concentration is determined from the amount of
absorption.
• Concentration measurements are usually determined from a working curve
after the instrument with standards of known concentration.

INSTRUMENTATION

• Parts of Atomic Absorption Spectrophotometer :

• Light source
• Nebulizer
• Atomizer
• Monochromator
• Detector and amplifier
• Read out system

LIGHT SOURCE

• Hollow cathode lamp are the most common radiation source in AAS.
• It contains a tungsten anode and a hollow cylindrical cathode .
• These are sealed in a glass tube filled with an inert gas. (mainly neon or
argon)
• Each elements has its own unique lamp which must be used for that analysis

NEBULIZER

• Nebulizer suck up liquid samples at controlled rate.

• Create a fine aerosol spray for introduction into the flame.
• Mix the aerosol and fuel and oxidant thoroughly for introduction into flame.
ATOMIZER

• Elements to be analysed needs to in atomic state and this is done by means

of atomizer.
• Atomization is separation of particles into individual molecules and breaking
molecules into atoms.
• This is done by exposing the analyte to high temperature in a flame or
graphite furnace.
• The atomizers most commonly used nowadays are (spectroscopic) flames
and electrothermal (graphite tube) atomizers.

FLAME ATOMIZATION

• Nebulizer suck up liquid sample at controlled rate and creates a fine aerosol
spray for introduction into flame.
• To create flame, we need to mix an oxidant gas and a fuel gas.
• In most of the cases air – acetylene flame or nitrous oxide acetylene flame is
used.
• Liquids or dissolved samples are typically used with flame atomizer.
• Steps in flame atomization :

ELECTRO THERMAL ATOMIZATION

• It uses a graphite coated furnace to vaporize the sample.

• Samples are deposited in a small graphite coated tube which then heated to
vaporize and atomize the analyte.
• The graphite tubes are heated using a high current power supply.
• Steps in electro thermal atomization : Drying Pyrolysis Atomization
Cleaning

MONOCHROMATOR

• This isvery important part in an AAS.

• It is used to separate out all of the thousand of lines.
• A monochromator is used to select the specific wavelength of light which is
absorbed by the sample and to remove other wavelengths.
• The selection of the specific light allows the determination of the selected
element in the presence of others.
DETECTOR AND AMPLIFIER

• The light selected by the monochromator is directed onto a detector whose

function is convert the light signal into an electrical signal.
• Photomultiplier tube detector is mainly used.
• The processing of electrical signal is fulfilled by a signal amplifier.
• The amplified signal is then displayed on read out system or fed into a data
station for printout by the requested format.

CALIBRATION CURVE

• A calibration curve is used to determine the unknown concentration of an

element in a sample.
• The instrument is calibrated using several solutions of known
concentrations.
• The absorbance of each known solution is measured & then a calibration
curve of concentrations vs absorbance is plotted.
• The sample solution is fed into instrument & the absorbance of the element
in the solution is measured.
• The unknown concentration of element is then calculated from the
calibration curve.

INTERFERENCES IN ATOMICABSORPTION SPECTROSCOPY

• Interference is a phenomenon that leads to change in intensity of analyte

signal in spectroscopy.
• Interferences in AAS fall into two basic categories :

1. Non-Spectral Interferences affect the formation of analyte items.

2. Spectral Interferences :

high light absorption due to presence of absorbing species

• Matrix interference
• Chemical interference
• Ionization interference
NON-SPECTRAL INTERFERENCES

• Matrix interferences :
• When a sample is more viscous or has different surface tension than the
standard it results in difference in sample uptake rate due to changes in
nebulization efficiency.
• Such interferences are minimized by matching the matrix composition of
standard and sample
• Chemical interferences :
• If a sample contains a species which forms a thermally stable compound
with analyte that is not completely decomposed by the flame energy then
chemical interferences exist.
• Such interferences are minimized by using higher flame temp. to provide
higher dissociation energy.

IONIZATION INTERFERENCE

• Itis more common in hot flames.

• The dissociation process doesn’t stop at formation of ground state atoms.
• Excess energy of the flame lead to excitation of ground state atoms to ionic
state by loss of electrons thereby resulting in depletion of ground state
atoms.
• Ionization interference is eliminated by an excess of an element which is
easily ionized thereby creating a large number of electrons in the flame &
suppressing the ionization of the analyte.

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 of these interferences are due to molecular emission from oxides of
other element is a sample.

APPLICATIONS OF ATOMICABSORPTION SPECTROSCOPY

 • Determination of small amount of metals (lead, mercury, calcium,
magnesium)
• AAS is widely used in metallurgy, alloys and in inorganic analysis.
• Biochemical Analysis : A number of elements present in biological samples
can be analysed by AAS. These include estimated of sodium, calcium,
potassium, zinc, iron, lead, mercury, etc.
• Pharmaceutical Analysis : Estimation of zinc in insulin preparation, calcium
in calcium salt is done by using AAS.

• Sodium, potassium, calcium in saline and ringer solution are estimated by

AAS.
• Analysis of ash for determining the content of sodium, potassium, calcium,
magnesium and iron is done by AAS.
• Atomic absorption spectroscopy is used in assay of
a) Intraperitoneal dialysis of fluid for calcium & magnesium.
b) Activated charcoal for zinc.
c) Cisplastin for liver.

Advantages of Atomic Absorption Spectrophotometer

• It has high sensitivity and can detect very small amounts of metals.
• It can detect about 70 elements.
• Only a small sample is required for analysis.
• The technique is rapid and gives quick results.
• It gives good accuracy for quantitative estimation.
• It is useful in many fields such as clinical, pharmaceutical, environmental,
and metal analysis.

Disadvantages of Atomic Absorption Spectrophotometer

• A trained person is required to operate it properly.
• The instrument is costly.
• A separate hollow cathode lamp is needed for each element, so the scope is
limited.
• It is not suitable for elements that form oxides on heating such as Al, W,
Mo, Ti, Si, and V.
• The sample usually must be in liquid form.
• Common anions like chloride, sulphate, and carbonate can affect the
analysis.
1) Atomic absorption spectrophotometer vs spectrophotometer

Atomic absorption
Spectrophotometer
spectrophotometer

Measures absorption by molecules or

Measures absorption by free atoms.
solutions.

Uses a hollow cathode lamp as Uses continuous light source like tungsten
light source. or deuterium lamp.

Needs atomizer to convert sample

Does not need atomization.
into atoms.

Used mainly for metal analysis. Used for general chemical analysis.

Very element-specific. Less element-specific.

Example: determination of lead, Example: UV-visible analysis of colored

iron, zinc. compounds.

2) Atomic absorption spectroscopy vs atomic emission spectroscopy

Atomic absorption spectroscopy Atomic emission spectroscopy

Measures absorption of radiation by Measures emission of radiation by excited

ground-state atoms. atoms.

Requires external light source. Does not require external light source.

Atoms absorb energy and move to Atoms are excited first, then emit light while
higher level. returning to lower level.

Usually more sensitive for trace

Often used for multi-element analysis.
metals.

Signal decreases with absorption. Signal increases with emission.

 Atomic absorption spectroscopy Atomic emission spectroscopy

Example: flame AAS. Example: flame photometry, ICP-OES.

3) Classical method vs instrumental method

Classical method Instrumental method

Based on instrumental measurement of physical

Based on chemical reactions.
properties.

Usually slower. Usually faster.

Less sensitive. More sensitive.

Requires more sample and

Requires small sample.
reagents.

More human error. Less human error.

Example: titration, gravimetry. Example: AAS, UV-Visible, HPLC.