Atomic Absorption Spectroscopy

SUMMER 2020

Professor Dr. Hasina Yasmin

Department of Pharmacy
 Atomic absorption spectroscopy
• Atomic absorption spectroscopy is a technique for determining the
concentration of a particular metal element within the sample.

• The technique was introduced in 1955 by Walsh in Australia. Atomic absorption

spectroscopy can be used to analyze over 60-70 different metals including most
of the common rare earth metals in a concentrations that range from trace to
micro quantities. Each element absorbs only radiation of well defined
wavelength, characteristic of the particular element.
• The basic idea of this technique is based on energy absorption during
transition of electrons from one energy level to another energy level. The
lowest energy electronic configuration of an atom is known as ground state.
When energy is provided, outer shell electrons are promoted to higher energy
excited state.
 Atomic Spectroscopy
Energy
n=1
DE
• Electron excitation n=2
• The excitation can occur at different degrees n = 3,
etc.
• low E tends to excite the outmost e-’s first
• when excited with a high E (photon of
high v) an e- can jump more than one
levels
• even higher E can tear inner e-’s away
from nuclei
• An e- at its excited state is not stable and 4f
tends to return its ground state 4d
n=4
4p
• If an jumped more than one energy levels
e- 3d
because of absorption of a high E, the 4s

Energy
process of the e- returning to its ground state n=3 3p
may take several steps, - i.e. to the nearest 3s
low energy level first then down to next … n=2 2p
2s
n=1 1s
 Atomic Spectroscopy
• The level and quantities of energy supplied to excite e-’s can be measured & studied in terms
of the frequency and the intensity of an e.m.r. - the absorption spectroscopy
• The level and quantities of energy emitted by excited e-’s, as they return to their ground state,
can be measured & studied by means of the emission spectroscopy
• The level & quantities of energy absorbed or emitted (v & intensity of e.m.r.) are specific for a
substance
• Atomic spectra are mostly in visible regions
 Atomic spectroscopy

• Atomic absorption
• Bright background (noise)
• Measure intensity change
• More signal than emission
• Trace detection
• Atomic emission
• Zero background (noise)
• Since each electron can show various
transitions between different energy
levels, hence there are many lines for
one electron system.
Atomic spectrum of hydrogen
Principle of Atomic absorption spectroscopy

• If light of just the right wavelength impinges on a ground state atom, the atom
may absorb light and enters into the excited state by a process called atomic
absorption.

• At the same time there will be a reduction in light intensity which is measured
and directly correlated with the concentration of the element. Light absorption of
unknown sample is then compared with well calibrated standard.
 Atomic Absorption Spectrometer
∆ ∆ ∆ ∆
Sample (aq. solution) molecules atoms atoms*
X ions

Detector

Recorder
Atomic Absorption Spectrometer……….
• Components of atomic absorption spectrometer are-

1. Radiation/light source
2. Atomizer
3. Monochromator
4. Detector
5. Readout device
Images courtesy:
• Chemguide
• PubChem
• Wikipedia
• jsanalytical
1. Radiation/light source
• Laser
• Hollow cathode lamp (HCL)
The common light source is hollow cathode lamp. It consists of a tungsten anode and a cylindrical
hollow cathode made up of the element to be determined. These are sealed in a glass tube, filled
with an inert gas, example: neon or argon.

The cathode contains the element that will be

analysed.
• Light emitted by hallow-cathode lamp has the
same wavelength as the light absorbed by the
analyte element.
• Different lamp required for each element (some
are multi- element)
 Wide range of WL

Narrow WL

Na
Steps of emitting radiation by HCL:
• Ionization: The inert gas atoms are ionized by applying a potential difference of
300-400V between anode and cathode.

• Sputtering: The gaseous ions bombard with cathode and eject metal atoms from
cathode in a process called sputtering
• Excitation: Collision of sputtered metal atoms with the gaseous ions results in
excited metal atoms

• Emission: The excited metal atoms then emit radiation characteristics of the
metal as they fall back to ground state.
2. Atomizer
• The function of atomizer is to convert the molecules of the liquid sample into
free gaseous atoms.
• High-temperature source such as a flame or graphite furnace are used as
atomizer
• Flame atomic absorption spectroscopy
• Graphite furnace atomic absorption spectroscopy
• The most common atomizer is Flame.

• The flame breaks the liquid sample into droplets which are then evaporated and
leave a solid residue. The residue is decomposed to give free atoms. These free
atoms then absorb radiation characteristic for that particular metal.
Flame Atomization

Nebulization - Conversion of the liquid

sample to a fine spray.

Desolvation - evaporation of the

solvent

Volatilization - Solid atoms are

converted to a vapor in
the flame.

• There are three types of particles that

exist in the flame:
1) Atoms
2) Ions
3) Molecules
 Laminar Flow Burners

• Sample is “pulled” into the

nebulization chamber by the flow
of fuel and oxidant.

• Contains spoilers (baffles) to allow

only the finest droplets to reach the
burner head.

• Burner head has a long path length

and is ideal for atomic absorption
spectroscopy.
Laminar Flow Burners

Nebulization
❑ Sucks up the liquid sample. (Aspiration)
❑ Creates a fine aerosol (fine spray) for introduction into the flame
❑ Mixes aerosol, fuel and oxidant thoroughly, creates a heterogeneous mixture
❑ the smaller the size of the droplets produced, the higher the element sensitivity
Types of Flames

Fuel / Oxidant Temperature

H-CC-H acetylene / air 2100 °C – 2400 °C (most common)
acetylene / N2O 2600 °C – 2800 °C
acetylene / O2 3050 °C – 3150 °C
• Selection of flame type depends on the volatilization temperature of the atom of interest.

Flame Structure

• Interzonal region is the hottest part of the flame and

best for atomic absorption.
• Fuel rich flames are best for atoms because the
likelihood of oxidation of the atoms is reduced.
• Oxidation of the atoms occurs in the secondary
combustion zone where the atoms will form molecular
oxides and are dispersed into the surroundings.
Graphite furnace atomic absorption spectroscopy (GFAAS)
➢ Sample holder: graphite tube
➢ Samples are placed directly in the graphite furnace
which is then electrically heated.
➢ Beam of light passes through the tube.
➢ Three stages:
1. Drying – for evaporation of solvent/moisture of
sample
2. Ashing – for removal of volatile hydroxides, sulfates,
carbonates
3. vaporization- atomization of analyte

Typical conditions for Fe in GFAAS:

Drying: 125º C for 20 sec
Ashing: 1200º C for 60 sec
Vaporization: 2700º C for 10 sec
Advantages of GFAAS over FAAS:
• Solutions, slurries and solid samples can be directly analyzed.
• Much more efficient atomization
• Greater sensitivity (ppb)
• Smaller quantities of sample (typically 5 – 50 µL) is needed

Disadvantages
• Expensive
• Requires high level of operator skill
• Not very reproducible (precision = 5-10% RSD)
• Requires detailed optimization procedures for every sample to obtain accurate results
• Frequent calibration is required
3. Monochromator
• Isolation of the absorption line from background light and from molecular
emissions originating in the flame, i.e. tuned to a specific wavelength
• Multi-element lamps: large number of emitted lines; isolation of the line of
interest

4. Detector
The detector used is a photomultiplier
tube. Its function is to measure the
intensity of radiation before and after
absorption by the sample. From this, we
can calculate how much radiation has
been absorbed from the intense beam.
The detection limit is the smallest amount of an element that can be reliably measured. Smaller
limits of detection (LODs) are better. Some common light metals have a lower LOD using flame
atomic emission. Most transition elements have a significantly lower LOD using AAS.

Detection limits (ppm = µg/mL)

Element Flame AAS Flame AES
Ag 3 20
Ca 1 0.1
Cd 1 2000
Fe 6 50
K 2 3
Mg 0.2 5
Mn 2 15
Na 0.2 0.1
Ni 3 600
Pb 5 200
Zn 1 200
Determination of Fe

1.00 mL pipette

1 mL 2 mL 3 mL 4 mL 5 mL

Fe:
0.05
mg mL-1
50.00 mL volumetric flasks
Determination of Fe
Calibration curve for absorbance of Fe

1.5
Absorbance

1 y = 0.2093x + 0.001

0.5

0
0 1 2 3 4 5 6

Fe concentration / ppm
 Applications of atomic absorption spectroscopy

1. Clinical analysis: Analyzing metals in biological fluids like blood and

urine etc.
2. Environmental analysis: Finding out the levels of various elements in
rivers, sea water, drinking water, air, petrol etc.
3. Pharmaceuticals: In pharmaceutical manufacturing process minute
quantities of a catalyst is often required which may be present in the
final product. Besides, there are pharmaceuticals containing metals
(e.g.: antacids, TPN etc.)
4. Industry: Raw materials are examined by atomic absorption
spectroscopy to check the presence of major elements and often toxic
impurities.
5. Mining: By using atomic absorption spectroscopy, the amount of gold
in rocks for example can be done.
 Advantages of atomic absorption spectroscopy

1. Simplicity: The principle of measurement is simple

2. Wide spread application: Atomic absorption spectroscopy is used widely for quantitative
determination of most of the metals
3. Sensitivity: Metals can be analyzed at a level of ppm (mg/L) and even ppb
4. Interference: It is possible to determine a metal in the presence of other metals without any
interference
5. The technique is applicable over a wide range of concentrations for most elements
6. The technique is relatively inexpensive
7. The equipment is easy to use
Interferences
-Spectral – absorption by other species than the analyte atoms
-non-spectral – affect the formation of analyte atoms
Non-spectral interference
1. Chemical Interference: Formation of stable or refractory compounds
refractory: elements that form stable compounds that are not completely atomized at the
temperature of the flame or graphite furnace

For example:
Calcium in the presence of phosphate forms stable calcium phosphate
3 Ca2+ + 2 PO43- Ca3(PO4)2
Solutions

•higher flame temperature (nitrous oxide/ acetylene instead of air/ acetylene)

•chelating agent or release agents can be added to the sample to protect or
free the analyte:
Addition of a chelating agent for the analysis of calcium:

Ca3(PO4)2 + 3 EDTA 3 Ca(EDTA) + 2 PO43-

Addition of a release agent for the determination of calcium:

for example: addition of 1000 ppm LaCl3

Ca3(PO4)2 + 2 LaCl3 3 CaCl2 + 2 LaPO4

2. Ionization Interference
If additional energy is applied, the ground state atoms can be transferred to the excited state or
ionized due to removal of an electron.
Problem
Alkali metals and alkaline earth metals have lowest ionization energies and are therefore most
easily ionized in flames.
Example: at 2450 K, p = 0.1 Pa, 5 % Na is ionized and 33 % K is ionized
Ionization leads to reduced signal intensity, as energy levels of ions are different from those of the
parent ions.
Solution
Ionization of the analyte element can be suppressed by adding an element that is more easily
ionized. Ionization of the added element results in a high concentration of electrons in the flame.
Example:
Addition of 1000 ppm CsCl when analyzing for Na or K
Images courtesy:
• Chemguide
• PubChem
• Wikipedia
• jsanalytical