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PRESENTED BY: ANSHUL SHARMA
M.PHARM ANALYSIS
1

 Introduction
 History
 Principle
 Instrumentation
 Applications
CONTENTS
2

 Atomic absorption is a very common for detecting
metals and metalloids in a samples.
 It is very reliable and simple to use.
 It can analyse over 62 elements.
 It also measure the concentration of metals in the
sample.
INTRODUCTION
3

 The first atomic absorption spectrometer was built
by the CSIRO (Commonwealth Scientific and
Industrial Research Organisation) scientist Alan
Walsh in 1954.
HISTORY
4

 The technique uses basically the principle that free
atoms generated in an atomizer can absorb
radiation at specific frequency.
 AAS quantifies the absorption of ground state
atoms in the gaseous state.
 The atoms absorb ultraviolet or visible light and
make transition to higher electronic energy levels.
The analyte concentration is determined from the
amount of absorption.
PRINCIPLE
5

 Concentration measurement are usually from a
working curve after calibrating the instrument with
standards of known concentration.
 Atomic absorption is a very common technique for
detecting metals and metalloids in environmental
samples.
6

OPERATIONAL
PRINCIPLE OF AAS
7

 Source of light
 Chopper
 Nebulizer
 Atomizer
 Monochromator
 Detectors
 Readout
INSTRUMENTATION
8

9

10

11

12

13

 Hollow Cathode Lamp (HCL)
 Electrodeless Discharge Lamps
SOURCE OF LIGHT
14

HOLLOW CATHODE LAMP
15

 HCL is the most common radiation source in
AAS.
 It contains a tungsten anode and a hollow
cylindrical cathode made of the element to
be determined.
 These are sealed in a glass tube filled with
an inert gas (neon, argon).
 Each element has its own unique lamp
which must be used for that analysis.
16

 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
atom inside the bulb causing them to emit their
characteristic spectrum.
ELECTRODELESS
DISCHARGE LAMP
17

 They are typically much more intense and, in some
cases, more sensitive than comparable HCL. Hence
better precision and lower detection limits where an
analysis is intensity limited.
 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.
18

CONSTRUCTION OF ELECTRODELESS
DISCHARGE LAMP
19

NEBULIZER
20

FUEL AND OXIDANT USED FOR
FLAME COMBUSTION
21

There is no nebulization, 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.
ELECTROTHERMAL
EVAPORATOR
22

 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.
HYDRIDE GENERATION
TECHNIQUE
23

 FLAME ATOMIZERS
 ELECTROTHERMAL ATOMIZERS
ATOMIZER
24

 Flame is used to atomize the sample.
 Sample when heated is broken into its atoms.
 High temperature of flame causes excitation.
 Electrons of the atomized sample are promoted to
higher orbitals, by absorbing certain amount of
energy.
FLAME ATOMIZER
25

26

 Graphite furnace atomic absorption spectrometry
(GFAAS) (also known as Electro thermal Atomic
Absorption spectrometry (ETAAS)) is a type
of spectrometry that uses a graphite-coated furnace
to vaporize the sample.
 Instead of employing the high temperature of a
flame to bring about the production of atoms from
the sample and it is non-flame methods involving
electrically heated graphite tubes or rods.
ELECTROTHERMAL
ATOMIZERS
27

SIMPLE SCHEMATIC
DIAGRAM
28

• 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.
29
WORKING

 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.
30
WORKING

 Sample holder:
 Graphite tube: Samples are placed directly in the
graphite furnace which is then electrically heated.
 Beam of light passes through the tube
GFAAS
31

 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
32

 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).
ADVANTAGES OF GFAAS
33

 Expensive.
 low precision.
 low sample throughput.
 requires high level of operator skill
DISADVANTAGES OF GFAAS
34

 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.
APPLICATIONS OF GFAAS
35

MONOCHROMATOR
- Wavelength
selectors
- Produces
monochromatic light
Consists of:
1) Entrance slit
2) Diffraction grating
3) Exit slit
Diffraction gratings
are mostly used
rather than prisms
36

GRATINGS AND PRISM
37

 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
DETECTOR
38

39

INTERFERENCE
40

INTERFERENCES &
CONTROL MEASURES
NON SPECTRAL
Matrix
Method of
Standard
Additions
Chemical
add an excess
of another
element or
compound
which
will form a
thermally
stable
compound
with the
interferent
using a
hotter
flame.
Ionization
adding an
excess of
an
element
which
is very
easily
ionized
SPECTRAL
Background
Absorption
Continuum
Source
Background
Correction
Zeeman
Background
Correction
41

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.
SPECTRAL INTERFERENCE
42

 Formation of compound of low volatility
 Decrease in calcium absorbance is observed with increasing
concentration of sulfate or phosphate.
HOW TO OVERCOME
 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.
CHEMICAL INTERFERENCE
43

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)
IONIZATION INTERFERENCE
44

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).
ADVANTAGES AND
DIADVANTAGES
45

 Quantitative analysis.
 Qualitative analysis.
 Simultaneous multicomponent analysis.
 Determination of metallic element in biological
materials.
 Determination of metallic element in food industry.
 Determination of Ca, Mg, Na, K in blood.
APPLICATIONS
46

THANK YOU
47

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Atomic absorption spectroscopy

  • 1. PRESENTED BY: ANSHUL SHARMA M.PHARM ANALYSIS 1
  • 2.   Introduction  History  Principle  Instrumentation  Applications CONTENTS 2
  • 3.   Atomic absorption is a very common for detecting metals and metalloids in a samples.  It is very reliable and simple to use.  It can analyse over 62 elements.  It also measure the concentration of metals in the sample. INTRODUCTION 3
  • 4.   The first atomic absorption spectrometer was built by the CSIRO (Commonwealth Scientific and Industrial Research Organisation) scientist Alan Walsh in 1954. HISTORY 4
  • 5.   The technique uses basically the principle that free atoms generated in an atomizer can absorb radiation at specific frequency.  AAS quantifies the absorption of ground state atoms in the gaseous state.  The atoms absorb ultraviolet or visible light and make transition to higher electronic energy levels. The analyte concentration is determined from the amount of absorption. PRINCIPLE 5
  • 6.   Concentration measurement are usually from a working curve after calibrating the instrument with standards of known concentration.  Atomic absorption is a very common technique for detecting metals and metalloids in environmental samples. 6
  • 8.   Source of light  Chopper  Nebulizer  Atomizer  Monochromator  Detectors  Readout INSTRUMENTATION 8
  • 14.   Hollow Cathode Lamp (HCL)  Electrodeless Discharge Lamps SOURCE OF LIGHT 14
  • 16.   HCL is the most common radiation source in AAS.  It contains a tungsten anode and a hollow cylindrical cathode made of the element to be determined.  These are sealed in a glass tube filled with an inert gas (neon, argon).  Each element has its own unique lamp which must be used for that analysis. 16
  • 17.   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 atom inside the bulb causing them to emit their characteristic spectrum. ELECTRODELESS DISCHARGE LAMP 17
  • 18.   They are typically much more intense and, in some cases, more sensitive than comparable HCL. Hence better precision and lower detection limits where an analysis is intensity limited.  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. 18
  • 21.  FUEL AND OXIDANT USED FOR FLAME COMBUSTION 21
  • 22.  There is no nebulization, 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. ELECTROTHERMAL EVAPORATOR 22
  • 23.   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. HYDRIDE GENERATION TECHNIQUE 23
  • 24.   FLAME ATOMIZERS  ELECTROTHERMAL ATOMIZERS ATOMIZER 24
  • 25.   Flame is used to atomize the sample.  Sample when heated is broken into its atoms.  High temperature of flame causes excitation.  Electrons of the atomized sample are promoted to higher orbitals, by absorbing certain amount of energy. FLAME ATOMIZER 25
  • 27.   Graphite furnace atomic absorption spectrometry (GFAAS) (also known as Electro thermal Atomic Absorption spectrometry (ETAAS)) is a type of spectrometry that uses a graphite-coated furnace to vaporize the sample.  Instead of employing the high temperature of a flame to bring about the production of atoms from the sample and it is non-flame methods involving electrically heated graphite tubes or rods. ELECTROTHERMAL ATOMIZERS 27
  • 29.  • 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. 29 WORKING
  • 30.   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. 30 WORKING
  • 31.   Sample holder:  Graphite tube: Samples are placed directly in the graphite furnace which is then electrically heated.  Beam of light passes through the tube GFAAS 31
  • 32.   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 32
  • 33.   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). ADVANTAGES OF GFAAS 33
  • 34.   Expensive.  low precision.  low sample throughput.  requires high level of operator skill DISADVANTAGES OF GFAAS 34
  • 35.   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. APPLICATIONS OF GFAAS 35
  • 36.  MONOCHROMATOR - Wavelength selectors - Produces monochromatic light Consists of: 1) Entrance slit 2) Diffraction grating 3) Exit slit Diffraction gratings are mostly used rather than prisms 36
  • 38.   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 DETECTOR 38
  • 41.  INTERFERENCES & CONTROL MEASURES NON SPECTRAL Matrix Method of Standard Additions Chemical add an excess of another element or compound which will form a thermally stable compound with the interferent using a hotter flame. Ionization adding an excess of an element which is very easily ionized SPECTRAL Background Absorption Continuum Source Background Correction Zeeman Background Correction 41
  • 42.  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. SPECTRAL INTERFERENCE 42
  • 43.   Formation of compound of low volatility  Decrease in calcium absorbance is observed with increasing concentration of sulfate or phosphate. HOW TO OVERCOME  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. CHEMICAL INTERFERENCE 43
  • 44.  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) IONIZATION INTERFERENCE 44
  • 45.  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). ADVANTAGES AND DIADVANTAGES 45
  • 46.   Quantitative analysis.  Qualitative analysis.  Simultaneous multicomponent analysis.  Determination of metallic element in biological materials.  Determination of metallic element in food industry.  Determination of Ca, Mg, Na, K in blood. APPLICATIONS 46