Atomic Absorption Spectroscopy

Atomic absorption spectroscopy is a technique which studies absorption of

electromagnetic radiations in relationship to molecular structure. It is a technique
for measuring the concentration of various elements in the sample through their
absorption of light. It is a relatively simple and reliable technique which uses
absorption of optical radiation by free atoms for determining the contents of
different elements.
Atomic absorption spectroscopy is based on the principle that when a beam of
electromagnetic radiation is passed through a substance, the radiation may either
be absorbed or transmitted depending upon the wavelength of the radiation.

The absorption of radiation would bring about an increase in the energy of the
molecule. The energy gained by the molecule is directly proportional to the
wavelength of radiation.

The increase in the energy of the molecule leads to the electronic excitations where
electrons jump to higher energy levels. A particular wavelength that a given
molecule can absorb depends upon the changes in vibrational, or rotational or
electronic states.

When a monochromatic radiation of frequency v is incident on a molecule, the

molecule in the gaseous state E1E1 absorbs a photon of energy hνhν, it undergoes
a transition from lower energy level to higher energy level.

A detector is placed to collect the radiation after interaction with the molecule
which shows that intensity has reduced. With wide range of frequencies, the
detector shows the energy has been absorbed only from the frequency.

νν = (ΔE)h(ΔE)h

Therefore we obtain an absorption spectrum which is defined as a record of the

radiation absorbed by the given sample as a function of wavelength of radiation.

The energy difference between the levels is given as,

ΔEΔE = E2−E1E2−E1 = hνhν = hcλhcλ.

 Applications

 One of the major applications of atomic absorption spectroscopy is for the

determination of structural elucidation of various substances. This
technique is more beneficial as a very small quantity of substance is required
for analyzing.
 Atomic absorption spectroscopy can be used to analyze water for its metal
quantity if present.
 Atomic absorption spectroscopy is used in many pharmaceutical
manufacturing processes where small quantities of catalyst remain in the
final product.
 Atomic absorption spectroscopy can also be used in biological tissues such
as blood, liver, brain tissue, muscle tissue and fluids for analyzing metals.

Basic Working:-

In the atomic absorption spectrometer, the source of radiation in the spectrometer

is the tungsten filament emitting white light or hydrogen discharge lamp. The
radiation from the source is directed by some device (for example in this case a
mirror) on the sample. The radiation then passes through an analyser (the grating in
this case), which selects the frequency reaching the detector at any given time. The
signal from the detector passes then to a recorder which is attached to the analyser
so as to produce a trace of the absorbance of varying frequencies.
Hollow cathode lamps
Hollow cathode lamps (HCL) are the most common radiation source in LS AAS
Inside the sealed lamp, filled with argon or neon gas at low pressure, is a
cylindrical metal cathode containing the element of interest and an anode. A high
voltage is applied across the anode and cathode, resulting in an ionization of the fill
gas. The gas ions are accelerated towards the cathode and, upon impact on the
cathode, sputter cathode material that is excited in the glow discharge to emit the
radiation of the sputtered material, i.e., the element of interest. Most lamps will
handle a handful of elements, i.e. 5-8. A typical machine will have two lamps, one
will take care of five elements and the other will handle four elements for a total of
nine elements analyzed.
Flame atomizers

The oldest and most commonly used atomizers in AAS are flames, principally the
air-acetylene flame with a temperature of about 2300 °C and the nitrous
oxide[3] system (N2O)-acetylene flame with a temperature of about 2700 °C. The
latter flame, in addition, offers a more reducing environment, being ideally suited
for analytes with high affinity to oxygen.
Liquid or dissolved samples are typically used with flame atomizers. The sample
solution is aspirated by a pneumatic analytical nebulizer, transformed into
an aerosol, which is introduced into a spray chamber, where it is mixed with the
flame gases and conditioned in a way that only the finest aerosol droplets (<
10 μm) enter the flame. This conditioning process is responsible that only about
5% of the aspirated sample solution reaches the flame, but it also guarantees a
relatively high freedom from interference.
On top of the spray chamber is a burner head that produces a flame that is laterally
long (usually 5–10 cm) and only a few mm deep. The radiation beam passes
through this flame at its longest axis, and the flame gas flow-rates may be adjusted
to produce the highest concentration of free atoms. The burner height may also be
adjusted, so that the radiation beam passes through the zone of highest atom cloud
density in the flame, resulting in the highest sensitivity.
The processes in a flame include the stages of desolvation (drying) in which the
solvent is evaporated and the dry sample nano-particles
remain, vaporization (transfer to the gaseous phase) in which the solid particles are
converted into gaseous molecule, atomization in which the molecules are
dissociated into free atoms, and ionization where (depending on the ionization
potential of the analyte atoms and the energy available in a particular flame) atoms
may be in part converted to gaseous ions.
Background absorption and background correction

The relatively small number of atomic absorption lines (compared to atomic

emission lines) and their narrow width (a few pm) make spectral overlap rare; there
are only few examples known that an absorption line from one element will
overlap with another ] Molecular absorption, in contrast, is much broader, so that it
is more likely that some molecular absorption band will overlap with an atomic
line. This kind of absorption might be caused by un-dissociated molecules of
concomitant elements of the sample or by flame gases. We have to distinguish
between the spectra of di-atomic molecules, which exhibit a pronounced fine
structure, and those of larger (usually tri-atomic) molecules that don’t show such
fine structure. Another source of background absorption, particularly in ET AAS,
is scattering of the primary radiation at particles that are generated in the
atomization stage, when the matrix could not be removed sufficiently in the
pyrolysis stage.
All these phenomena, molecular absorption and radiation scattering, can result in
artificially high absorption and an improperly high (erroneous) calculation for the
concentration or mass of the analyte in the sample. There are several techniques
available to correct for background absorption, and they are significantly different
for LS AAS and HR-CS AAS.

Absorption spectroscopy refers to spectroscopic techniques that measure the

absorption of radiation, as a function of frequency or wavelength, due to its interaction
with a sample. The sample absorbs energy, i.e., photons, from the radiating field. The
transmitted energy can be used to calculate the absorption.

Atomic absorption is so sensitive that it can measure down to parts per billion of a
gram (µgdm–3) in a sample. The technique makes use of the wavelengths of light
specifically absorbed by an element.

CALCULATIONS
Calculation - Direct determination of liquid samples: Read the metal value in mg/l
from the calibration curve or directly from the readout system of the instrument.
 (1) If dilution of sample was required:
C + B
mg/l metal in sample = A x -------
B

A = mg/l of metal undiluted aliquot

from calibration curve
B = ml of deionized distilled water
used for dilution
C = ml of sample aliquot

(2) For samples containing particulate:

V
mg/l metal in sample = A x ---
C

A = mg/l of metal in processed sample

from calibration curve
V = final volume of the processed sample in ml
C = ml of sample aliquot processed

(3) For solid samples: report all concentrations as mg/kg dry weight

Dry sample:
A x V
mg metal/kg sample = -------
D

A = mg/l of metal in processed sample

from calibration curve
V = final volume of the processed sample
in ml
D = weight of dry sample in grams

Wet sample:

A x V
mg metal/kg sample = -------
W x P

A = mg/l of metal in processed sample

from calibration curve
 V = final volume of the processed sample
in ml
W = weight of wet sample in grams
P = % solids

Metals that can be Detected by Atomic Absorption Spectroscopy

Aluminium Al Copper Cu Mercury Hg
Antimony Sb Gallium Ga Molybdenum MoNickel Ni
Arsenic As Hafnium Hf Niobium Nb
Beryllium Be Indium In Ruthenium Ru
Barium Ba Iron Fe Tin Sn
Calcium Ca Lead Pb Tungsten W
Chromium Cr Lithium Li Vanadium V
Cadmium Cd Magnesium Mg Zinc Zn
Cobalt Co Manganese Mn Zirconium Zr

FOR COMPLETE PROCEEDURE , SEE THE MANUAL OF AAS IN THE

LAB