SUBJECT FORENSIC SCIENCE

Paper No. and Title PAPER No.4: Instrumental Methods and Analysis

Module No. and Title MODULE No.19: Atomic Absorption Spectroscopy

Module Tag FSC_P4_M19

FORENSIC SCIENCE PAPER No.4: Instrumental Methods and Analysis

MODULE No.19: Atomic Absorption Spectroscopy
 TABLE OF CONTENTS
1. Learning Outcomes

2. Introduction

3. Principle of Atomic Absorption Spectroscopy

4. Methodology

5. Instrumentation of Atomic Absorption Spectrophotometer

6. Functioning of the Atomic Absorption Spectrophotometer

7. Applications of Atomic Absorption Spectrophotometry

8. Sensitivity and Detection Limits of AAS

9. Summary

FORENSIC SCIENCE PAPER No.4: Instrumental Methods and Analysis

MODULE No.19: Atomic Absorption Spectroscopy
 1. Learning Outcomes
After studying this module, you shall be able to know about-

 The significance of Atomic Absorption Spectroscopy

 The basic principle and methodology of Atomic Absorption Spectroscopy
 The types and application of Atomic Absorption Spectroscopy

2. Introduction
Atomic absorption is a process involving the absorption of light by free atoms of an element
at a wavelength specific to that element. It is a means by which the concentration of metals
can be measured in their atomic state. In various phenomena of Atomic Spectroscopy
(emission, absorption and fluorescence), energy is supplied to the atoms in the form of
thermal, electromagnetic, chemical or electrical energy which is converted to light energy by
various atomic and electronic processes before measurement. Atomic Absorption
Spectrometry is useful not only for the identification but also for quantitative determination
of many elements present in samples. The technique is specific and sensitive in that
individual elements in each sample can be dependably identified in small amounts,

This technique was introduced for analytical purpose by Walsh and Alkemade under the
designation Atomic absorption spectroscopy. Walsh used the hollow cathode lamps as a line
source, greatly reducing the resolution required for successful analysis. As photomultipliers
had become accessible, the complications associated with measuring absorption with a
photographic plate were eliminated. His introduction of modulation into the system permitted
the detector to distinguish between absorption and emission by atoms at the same
wavelength. He also utilized the flame for atomization. It is found to be superior to other
technique as it can be used to determine 50-60 elements from trace to large quantities. These
may include metals and non-metals.

FORENSIC SCIENCE PAPER No.4: Instrumental Methods and Analysis

MODULE No.19: Atomic Absorption Spectroscopy
 3. Principle of Atomic Absorption Spectroscopy
The principle is based on the Beer- Lambert’s law. The absorption of radiation by the free
atoms is proportional to their concentration.

Or

Where,
Io = Intensity of incident radiation
It = Intensity of transmitted radiation
C = Concentration of analyte
K = Constant
L = Path length

4. Methodology
The sample solution is aspirated into the flame or heated in a tube to convert them into atoms
by the process known as atomization. The atoms then absorb radiations of characteristic
wavelength, promoting the electrons from ground state energy level to excited state. Each
element absorbs radiations of a particular wavelength which forms the basis for qualitative
analysis Depending on the number of atoms in the light path, the amount of light absorbed
also changes. By detecting this amount of light, a quantitative determination of the analyte
can be made.

FORENSIC SCIENCE PAPER No.4: Instrumental Methods and Analysis

MODULE No.19: Atomic Absorption Spectroscopy
 Each atom contains a positively charged nucleus, surrounded by a number of electrons in
rapid motion around the nucleus. For each electron in each atom, there is a discrete set of
energy levels that the electron can occupy. The spacing of the energy levels is different for
each electron in the atom, but for similar atoms corresponding electrons have identical
spacing. The energy levels are usually labelled E0, the ground state, through E1, E2 etc. to E∞.
For an unexcited atom, each electron is in the ground state. To excite the atom, one or more
electrons can be raised to the first or higher energy levels by the absorption of energy. This
energy can be supplied by photons or by collisions due to heat. Those electrons furthest from
the nucleus require least energy to go from the ground state E0 to the first energy level E1.
The energy E corresponds to the energy gap between the ground state and the first energy
level

E = E1 - E0
The energy required for this transition can be supplied by a photon of light with an energy
given by:
E=hν
Where, h is the Planck's constant and ν the frequency.

This corresponds to a wavelength (λ) of:

λ = h c/ E
Where, c is the speed of light in vacuum.

5. Instrumentation of Atomic Absorption Spectrophotometer

An Atomic Absorption Spectrophotometer apparatus consists of:

5.1 Radiation source:

The common source of light is a Hollow Cathode Lamp (HCL). This contains a tungsten
anode and a cylindrical hollow cathode made of the element to be determined. These are
sealed in a glass tube filled with an inert gas e.g., neon or argon at a pressure of between 1
N/m2 and 5 N/m2. The ionisation of some gas atoms occurs by applying a potential difference
of about 300 – 400 V between the anode and the cathode. These gaseous ions bombard the
cathode and eject metal atoms from the cathode in a process called sputtering. Some
sputtered atoms are in excited states and emit radiation characteristic of the metal as they fall
back to the ground state. These emitted radiations form incident radiations for the element
under analysis.

FORENSIC SCIENCE PAPER No.4: Instrumental Methods and Analysis

MODULE No.19: Atomic Absorption Spectroscopy
 5.2 Atomizer:

Nebulisation is the mechanism by which the sample solution is introduced as fine spray into
the flame. Nebulisation refers to the dispersion of a liquid into particles by a rapidly moving
gas, liquid stream or by mechanical means. This process is immediately followed by
atomization, wherein high energy like that of a flame converts molecules into atoms.

5.2.1 Flame Atomization: The burner and a nebulizer help in the atomization of the
element. Atomization occurs in the flame and an atomic vapour of the element to
be analysed is produced. The selection of flame temperature is important for
atomization. When it is low, atomization will be partial and when it is high, the
atoms may get ionized.

5.2.2 Graphite furnace atomization: A graphite furnace can be employed instead of

flame for atomization. The atomizer may be elongated along its axis to increase
the distance between the optical path and the sample deposition point which
increases the analytical sensitivity.

5.2.3 Oxidants and fuel: Fuels commonly used are hydrogen, propane, butane,
acetylene and natural gas. Oxygen and nitrous oxide are used as oxidants. The
oxygen - acetylene flame is most often used. Its temperature is high enough to
cause sufficient atomization for most elements which can be determined by AAS,
but not so high that ionization interference becomes significant. The analytical
conditions can be optimized by changing the composition of the gas mixture.

Table: Upper temperature limits for some gas mixtures

Combustible mixture Maximum temperature (K)

Butane – Air 2200
Acetylene – Air 2600
Acetylene - Nitrous Oxide 3100
Acetylene - Oxygen 3400

FORENSIC SCIENCE PAPER No.4: Instrumental Methods and Analysis

MODULE No.19: Atomic Absorption Spectroscopy
 5.3 Monochromator:

It is important that the instrument be capable of providing a narrow band width to separate
the line chosen for determination from other undesirable lines. Usually used devices are
gratings or prisms.

5.4 Detectors:

Photomultipliers are commonly used as detectors. In some instruments additional filters and
detectors are used to compensate for the fluctuations in the output of the source. The output
of photomultiplier is amplified which helps in source modification.

Block ray diagram of AAS

6. Functioning of the Atomic Absorption Spectrophotometer

Atomic Absorption Spectroscopy uses the absorption of light to measure the concentration of
gaseous atoms. The vaporization of the analyte ions or atoms is carried out in flame or
graphite furnace as the samples are generally solids or liquids. Transitions to higher
electronic energy level are made by the atoms as they absorb visible or ultra-violet light.
Determination of the analyte concentration is carried out by studying the amount of
absorption. In this process, a blank solution is sprayed into the flame and the meter is
adjusted for zero absorbance or 100% transmittance. Following this, the solution under
investigation is sprayed, the atoms in ground state absorb certain part of light resulting
decrease in transmitted light or increase in absorbed light falling on photomultiplier tube. The

FORENSIC SCIENCE PAPER No.4: Instrumental Methods and Analysis

MODULE No.19: Atomic Absorption Spectroscopy
 readings can be tabulated and with the help of standard graph, the concentration of a
particular element in the sample can be found out.

7. Applications of Atomic Absorption Spectrophotometry

Atomic absorption spectrometry has many uses in different areas of chemistry. The different
methods of Atomic absorption spectrometry are very powerful for analysis elements in a
solution. The instruments are simple and easy to operate. They are useful when few elements
have to be determined in a large number of samples, as is the case in clinical or food analysis.
Atomic absorption spectroscopy methods are of great importance compared with other
methods of elemental analysis. In clinical analysis, it is used for analysing metals in
biological fluids such as blood and urine.

In environmental analysis, it is utilized for monitoring the levels of various elements in rivers,
seawater, drinking water and air etc., Presence of contaminants can be monitored in food
stuff like fruit juices and wines. AAS can trace presence of pesticide residues in fruits and
vegetables. In pharmaceuticals industries, it is used for the assay of drugs. Purity of the
sample can be checked for minute quantities of a catalyst used in the manufacturing process
(usually a metal) sometimes present in the final product. Levels of the toxic substances
present in the products can be verified and reduced by this technique. By using Atomic
absorption spectrometry in mining industries, the amount of metals such as gold in rocks can
be determined to see whether it is worth mining the rocks to extract the gold.

8. Sensitivity and Detection Limits of AAS

More than seventy elements can be measured by Atomic Absorption Spectroscopy. The
lowest concentration that can be quantified in a sample depends upon many factors. In
spectrometry the sensitivity for an element is defined as being the concentration expressed in
µg/L which, in aqueous solution, leads to a 1% decrease A= 0.0044) in the transmitted light
intensity. So, for manganese (Mn), this value is 4pg with an aqueous matrix. When possible,
a calibration curve should be established with concentrations in the range of 20 to 200 times
this value. In practice, the concentrations must be at least ten times higher than the detection
limit to give reliable measurements.

FORENSIC SCIENCE PAPER No.4: Instrumental Methods and Analysis

MODULE No.19: Atomic Absorption Spectroscopy
 9. Summary

 This technique was introduced for analytical purpose by Walsh and Alkemade, under the
designation Atomic absorption spectroscopy. It is found to be superior to other
techniques as it can be used to determine elements from trace to large quantities.

 Atomic absorption spectroscopy (AAS) is an analytical measurement method relying on

the spectroscopic processes of excitation. It is used for the qualitative and quantitative
analysis of around seventy elements (metals or non-metals).

 In Atomic absorption spectroscopy, the absorption of resonant radiation by ground state

atoms of the analyte is used as the analytical signal. This process is highly selective as
well as very sensitive.

 The functioning of Atomic absorption spectroscopy is based upon the principles of

Lambert – Beer’s Law.

 The sample is first converted into an atomic vapour and then the absorption by the
atomic vapour is measured at a selected wavelength characteristic of atoms each element.
The amount of light absorbed is proportional to the concentration of the element.

 In its simplest form, an AAS resembles a single beam spectrophotometer which contains
four principal components. The light beam issuing from the source passing through the
burner in which the element is brought to its atomic state before being focused upon the
entrance slit of the monochromator which selects a very narrow wavelength interval. The
optical path ends at the entrance slit of the detector.

FORENSIC SCIENCE PAPER No.4: Instrumental Methods and Analysis

MODULE No.19: Atomic Absorption Spectroscopy