RESEARCH II QUARTER 3 Week 5

C.N: _________
NAME: ___________________________________________ GR. & SEC: _____________________

Common Laboratory Techniques 2

(AAS)
Learning Competencies:
The learner:
1. discuss the basic principles of atomic absorption spectroscopy;
2. discuss the use of AAS
3. understand the AAS application.

HOW TO USE THIS MODULE?

Read the simple instructions below to successfully meet the objectives of this kit.
1. Follow and read carefully all the contents and instructions indicated in every
page of this module.
2. Write on your notebook the concepts about the lesson. Writing enhances
learning.
3. Perform all the provided activities in the module.
4. Let your facilitator/ guardian assess your answers using the answer key card.
5. Analyze conceptually and answer the “Let’s Assess What You Have Learned”
part.
6. Enjoy studying!

PreTest
Directions: Write the letter that corresponds to the definition of the following terms
on the blank provided before the number.
____1. Atomic-absorption (AA) spectroscopy uses the absorption of light to measure the
concentration of _______-phase atoms.
A. solid
B. liquid
C. gas
D. vapor
____2. The incident light beam is attenuated by atomic vapor absorption according to ______
law.
A. Beer’s
B. Newton’s
C. Ohm’s
D. Coulomb’s
____3. It is the process of reduction of sample to ground state atoms by application of heat
by means of a flame or a graphite furnace.
A. Atomic Absorption Spectrometer
B. UV-VIS spectrometer
C. Infrared spectrometer
D. Fourier transformed infrared spectrometer
____4. The process of reduction of sample to ground state atoms by application of heat by
means of a flame or a graphite furnace in AAS.
A. absorbance
B. atomisation
C. aspiration
D. emission
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____5. The _________ is an important step in AAS as aids in determining the sensitivity of
the reading.
A. radiation source
B. atomization
C. spectrometer
D. absorption

LOOKING BACK
Spectroscopy
• It is the branch of science that deals
with the study of interaction of matter
with light. OR
• It is the branch of science that deals
with the study of interaction of
electromagnetic radiation with matter.
What is FTIR spectrometry?
FTIR spectrometers (Fourier Transform Infrared Spectrometer) are widely used in
organic synthesis, polymer science, petrochemical engineering, pharmaceutical industry and
food analysis. In addition, since FTIR spectrometers can be hyphenated to chromatography,
the mechanism of chemical reactions and the detection of unstable substances can be
investigated with such instruments.
What is UV-Visible spectrometry?
UV-Visible spectroscopy is also known as electronic spectroscopy. In which the amount of
light absorbed at each wavelength of UV and Visible region of electromagnetic spectrum is
measured. This absorption of electromagnetic radiation by the molecules leads to molecular
excitation
Directions: Final step of research process is the Interpretation of Data, Provide the Three
Areas of Interpretation of Data and briefly explain each area.

FTIR spectrometry UV-Visible spectrometry

1.

2.

3.

BRIEF INTRODUCTION

What is AAS/Atomic Absorption Spectroscopy? 1

Atomic Absorption Spectroscopy is an instrumental analysis technique for rapid trace metal
analysis. It is based on element specific
wavelength light absorption by ground state
atoms in the flame or electro thermal graphite
furnace.
It finds immense applications in the analysis for
trace metals in soils, lakes, rivers, oceans, and
drinking water, pharmaceuticals, foods and
beverages, geological and mineralogical samples,
petroleum products, biological fluids and
specimens and forensic analysis.

___________________
1 https://lab-training.com/aas/

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 ACTIVITY 1: “Atomic Absorption Spectroscopy”

The phenomenon of atomic absorption (AA) was first observed in 1802 with the
discovery of the Fraunhofer lines in the sun's spectrum. It was not until 1953 that Australian
physicist Sir Alan Walsh demonstrated that atomic absorption could be used as a
quantitative analytical tool. Atomic absorption analysis involves measuring the absorption of
light by vaporized ground state atoms and relating the absorption to concentration. The
incident light beam is attenuated by atomic vapor absorption according to Beer's law.

Atomic-absorption (AA) spectroscopy

uses the absorption of light to measure
the concentration of gas-phase atoms.
Since samples are usually liquids or
solids, the analyte atoms or ions must
be vaporized in a flame or graphite
furnace. The atoms absorb ultraviolet
or visible light and make transitions to
higher electronic energy levels. The
analyte concentration is determined
from the amount of absorption. Applying the Beer-Lambert law directly in AA spectroscopy
is difficult due to variations in the atomization efficiency from the sample matrix, and
nonuniformity of concentration and path length of analyte atoms (in graphite furnace AA).
Concentration measurements are usually determined from a working curve after calibrating
the instrument with standards of known concentration.2

Uses of Atomic Absorption Spectroscopy

• Atomic Absorption Spectroscopy provides cost-effective viable solutions for the analysis
of trace amounts of metals in the entire range of natural and manmade materials such
as Geological samples, Environmental samples, Biological Specimens, Agricultural
produce and soils, Pharmaceuticals, Foods and Drinking water.

• The technique affords advantages of speed, sensitivity and precision over the classical
gravimetric methods. Introduction of accessories such as graphite furnace, flow
injection analysis and improvements in the suppression of matrix interferences have
further contributed to improvement in sensitivity and selectivity of analytes in complex
matrices.

• Atomic Absorption Spectroscopy applications in the field of environment, drinking

water, mining and mineralogy, oceanographic studies, soils, pharmaceuticals, foods,
toys, forensic investigations are of great significance.

• The list is endless and presence or absence of trace metals is a factor that cannot be
overlooked for evaluation of characteristics of materials or concerns regarding human
health and safety.

• The chemical techniques used for the analysis of trace metals have evolved from simple
gravimetric methods to highly sophisticated time saving instrumental techniques.
Atomic Absorption Spectroscopy is a popular technique which involves moderate
investment and affordable operational cost.

• These features coupled with a high degree of accuracy and precision of results has
contributed to the widespread presence of atomic absorption spectrometers in college
laboratories, industrial laboratories and regulatory body laboratories across the world.

_______________________
2 https://web.nmsu.edu/~esevosti/scheme.htm

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Principle of Atomic Absorption Spectroscopy

Atomic absorption spectroscopy (AAS) is based upon the principle that free atoms in
the ground state can absorb light of a certain wavelength. These very specific wavelengths
give the technique excellent specificity and detection limits in the AAS analysis. Absorption
for each element is specific, no other elements absorb this wavelength. Typical applications
of AAS include –
• Quantitative metal concentrations in solution
• Analysis of lead in paint
• Monitoring of trace metals in industrial effluent streams
• Trace elements in product/raw materials along with ICP-MS
• Analysis of additives and purity in steels and other metal alloys
• Analysis of low-level contaminants

Several analytical techniques have been applied for detection and quantitative
estimation of trace metals in different types of matrices. Classical techniques based on
gravimetric and titrimetry provided good accuracy but were time-consuming.

Increasing demand for high speed analysis led to the introduction of instrumental
methods such as Ion selective electrodes, UV-VIS spectroscopic techniques, Atomic
Absorption Spectroscopy, ICP – OES (Inductively coupled plasma - optical emission
spectrometry) and ICP – MS (Inductively coupled plasma mass spectrometry). The choice of
technique depends on the required detection levels, available sample quantity and most
important available budget. The topic is covered to some extent in the article which elemental
analysis technique is right for me.

Atomic Absorption Spectroscopy is a moderately priced instrumental analysis

technique which provides a high degree of accuracy and precision of results. Due to its high
analysis throughput, it finds its rightful place in university laboratories, pollution control
laboratories and industrial quality control laboratories.
The present article highlights some areas where an awareness of working with an
Atomic Absorption Spectrometer will prove to be an asset in enhancing your professional
growth.
In case you are engaged in any of the activities or areas discussed in the article or wish
to land into such areas, you will stand to gain through up-gradation of your knowledge and
technical skills on this technique.

Activity 1: a. Image Intensifier. Surround with the appropriate words and phrases the
expression in the middle of the graph.

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b. Fill in the blank with the correct answer

The phenomenon of atomic absorption (AA) was first observed in 1802 with the
discovery of the _______________ in the sun's spectrum. It was not until ______ that Australian
physicist _____________ demonstrated that atomic absorption could be used as a quantitative
analytical tool. Atomic absorption analysis involves measuring the _____________ by vaporized
ground state atoms and relating the absorption to concentration. The incident light beam is
attenuated by atomic vapor absorption according to _________ law.
Atomic-absorption (AA) spectroscopy uses the ________ of light to measure the
concentration of _________ atoms. Since samples are usually liquids or solids, the analyte
atoms or ions must be vaporized in a flame or ___________. The atoms absorb _________ or
visible light and make transitions to higher electronic energy levels.
Atomic absorption spectroscopy (AAS) is based upon the principle that ____ atoms in
the ground state can absorb light of a certain _________.

ACTIVITY 2: Learning Atomic Absorption Spectroscopy

Atomic Absorption Spectroscopy Applications

• Mining and Geology – The elemental composition of minerals and rocks provide
valuable information on the commercial feasibility of conducting mining activities in
areas explored. After mining, the ores and minerals need to be tested for composition
for the efficiency of refining operations. Similarly, trace metal analysis is of great value
in prospecting for oil and water deposits.
Gemstones are also graded on the basis of the presence of certain trace metals.
Elemental composition of archaeological artefacts is helpful for tracing their source.

• Environmental Monitoring – Environmental monitoring for trace metal

contamination of industrial effluents, oceans, rivers and lakes is important for
establishing the safety of water for drinking and commercial use. It is important to
establish if such samples are within the safety limits set by regulatory bodies.
Environmental monitoring also plays a significant role in the evaluation and feasibility
of the site for setting up commercial projects.

• Materials Development – Common properties of materials such as hardness,

brittleness, grain size, crystallinity and amorphous nature are significantly influenced
by composition and trace metals. Trace metal analysis can provide useful information
on the performance properties of such materials.

• Pharmaceuticals – Trace metal analysis plays an important role in formulation

development, catalyst efficiency and dosage limits. Most elements have a beneficial role
up to certain prescribed limits but beyond such limits the effects are harmful.

• Foods and Beverages – In synthetic processed foods, metal pickup takes place due to
contact with processing equipment and catalytic conversions. Consumer awareness on
food safety is increasing by the day so manufacturers have to ensure that the trace
metals do not exceed the permissible limits and this requires rigorous quality control
through atomic absorption spectroscopy and other sophisticated instruments.

• Oil and Petroleum – Both edible oils and mineral oils require refining before
consumption. Such refining operation can involve distillation as well as catalytic
refining. Uptake of metals during such operations can lead to deterioration of
performance or consumer hazards. Trace metal analysis of engine oil provides useful
diagnostic information on the wear and tear of engine parts.

• Agriculture – Trace metal constitution of soils in addition to their acidic or basic

nature is essential to establish their productivity and nutrient value. Trace metal

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 composition of plants (leaves, stems and roots) gives a fair idea on how the uptake of
minerals gets distributed under different growth conditions

• Forensics – Trace metal analysis provides valuable information on specimens such as

stomach contents for food poisoning, paint chips, fibres and hair strands collected
from the scene of a crime.

Types of Atomic Absorption Spectroscopy

Nowadays, Atomic Absorption Spectrometry (AAS) systems are comparatively

inexpensive instruments. Some also predict multi(few)-element capability.
There are various types of AAS – Flame (F AAS- Flame atomic absorption
spectroscopy ), Cold vapour (CV AAS- Cold Vapor Atomic Absorption Spectroscopy), Hydride-
generating (HG AAS - Hydride generation atomic absorption spectroscopy), and Graphite
furnace (GF-AAS - Electrothermal or graphite furnace atomic absorption spectrometry)
systems.

Instrumentation of AAS

• Atomizer
The sample must be atomized first in order to be studied. Atomization is an important
step in AAS as aids in determining the sensitivity of the reading. An effective atomizer
creates a large number of homogenous free atoms. Though there are many types of
atomizers present, only two are commonly used: Flame and Electrothermal atomizers.

• Radiation Source
There is a radiation source which irradiates the atomized sample. The sample absorbs
some of the radiation, and the rest passes through the spectrometer to a detector.
Radiation sources are of two categories: Line sources and Continuum sources. Line
sources excite the analyte and thus emit its own line spectrum. Continuum sources
have radiation that spreads out over a wider range of wavelengths.

• Spectrometer
Spectrometers are used to differentiate between various types of wavelengths of light
before they pass to the detector. The spectrometer in AAS can be either single-beam or
double-beam.

Single-beam spectrometers require radiation to pass directly through the atomized

sample. Whereas, double-beam spectrometers require two beams of light – one beam
that passes directly through the sample, and another that does not pass through the
sample at all.

Learning Atomic Absorption Spectroscopy

• Understanding the basics and operation of Atomic Absorption Spectroscopy is a career
objective of every trace metal analyst. Today’s trace metal analyst cannot afford to
remain ignorant of this well-established technique.

• Awareness and need for testing of materials existed even in ancient times and has
grown to keep its pace with the growth of human civilization. Today you cannot imagine
any man-made product be it a machine tool, glass decorative item, food product,
pharmaceutical, plastic ware or for that matter any other product which has not
undergone quality control using analytical techniques at some stage of its
manufacture.

• Even our natural resources such as water, air, food grains, fruits and vegetables are
certified for human consumption after undergoing laboratory testing.

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 • Analysis of trace metals gained significance on the onset of the age of metals. Even in
those times, it was common knowledge that composition of alloys has a bearing on
properties of metals to be used for the development of weapons for warfare, hunting,
implements, storage of foods and drinking water.

• Efficacy of herbal medicines based on ancient systems such as Ayurveda, Unani and
Siddha is dependent on the presence of trace metals or their oxides incorrect amounts.
An excess of such components could be disastrous to the consumer.

• Knowledge of AAS, its potential applications and operational aspects is an asset for
any analytical scientist.

Glossary of AAS terms

The glossary will help you to understand the terminology, below are some terms used in AAS.

Atomic Study of element specific light absorption by ground state atoms for estimation
Absorption of concentration of the element in the sample solution.
Spectroscopy
Atomisation Process of reduction of sample to ground state atoms by application of heat by
means of a flame or a graphite furnace.
Atom The smallest particle of an element or compound. It comprises of a central
nucleus containing neutral particles called neutrons and positively charged
protons. The electrons revolve the central nucleus in shells of different energy
levels. The number of electrons equals the number of protons in the neutral
atom.
Atomic Qualitative identification and quantification of element by emission of
Emission characteristic wavelength of light on excitation of an element by means of a
Spectroscopy flame or plasma
Atomic Measurement of light emitted on decay of elements from excited states.
Fluorescence Measurement is made at an angle to the optical beam path so that the detector
Spectroscopy sees only the fluorescence in the flame and not incidental light from the lamp.
Absorbance The amount or fraction of incident light absorbed by the ground state atoms. It
is directly proportional to the number of ground state atoms in the beam path
and also on the optical path length of the flame in accordance to Beer Lambert
law of light absorption
Absorbance unit a ratio of intensity of transmitted flight to the intensity of incident light. It is a
unit less quantity but is commonly expressed in absorbance units (EU)
Aspiration losses of reduction of liquid sample stream into fine droplets for introduction
into the flame
Acetylene Commonly used gas as fuel to support combustion of the flame. Provides
temperatures in the range 2150-23000C
Argon Gas used commonly as a filling gas in hollow cathode lamps and as sample
carrier in graphite furnace analysis
Air Used as oxidant in combination with acetylene as fuel gas to support the flame
Air compressor Device for delivery of air to the atomic absorption spectrometer. Oil less air
compressor is preferred as contamination from oil is thereby avoided
Burner A component of AAS system made of solid metal body having slit on the
flat top surface to provide the flame required for atomisation of the sample
Blaze angle It is the angle of cut of a mechanically ruled grating at which the angle of
incidence is equal to the angle of reflection so that light intensity is greatest
with minimal loss due to diffraction. For greater efficiency dual blazed ratings
are used which provide greater light throughput over the wavelength range of
the spectrometer
Collimation Condensation of beam of light as per size requirement
Cathode An electrode inside the lamp made from the pure metal whose analysis is
required in the sample solution
Detector A component of the system that records the intensity of the transmitted light.
Photomultiplier tube is the commonly used detector in AAS
Excitation Excitation of a ground state atom to higher energy states by means of
electromagnetic radiation
Grating A light dispersing device used in the monochromator
Hollow cathode A light source used for AAS analysis which is specific for metal to be analysed
lamp in the sample. For some elements multielement hollow cathode camps are also
used
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 Fraunhofer are a set of spectral absorption lines named after the German physicist Joseph
lines von Fraunhofer. The lines were originally observed as dark features in the
optical spectrum of the Sun.
Graphite A graphite tube about a cm long with a hole on top for atomisation of sample
furnace using electrical heating of the tube
Monochromator A device used for dispersion of incident light using prism or grating, reflecting
mirrors and a combination of entrance and exit slits for isolation of required
wavelength and collimation of the light beam

Activity 2:
Concept Learning Assessment:
Directions: Do a short research of the application AAS.
A. One specific application (ex. Forensics)
B. brief discussion

POST TEST

Directions: Write the letter that corresponds to the definition of the following terms
on the blank provided before the number.

____1. A light source used for AAS analysis which is specific for metal to be analysed in the
sample.
A. burner
B. blaze
C. Hollow cathode lamp
D. Hydride generation technique
____2. It is the amount or fraction of incident light absorbed by the ground state atoms in
AAS.
A. Aspiration
B. Absorbance
C. Collimation
D. Excitation
____3. It is an electrode inside the lamp of AAS made from the pure metal whose analysis is
required in the sample solution
A. Anode
B. Electron
C. cathode
D. anion
____4. It is the commonly used gas as fuel to support combustion of the flame in AAS.
A. Helium
B. Nitrogen
C. Argon
D. Acetylene
____5. Gas used commonly used as a filling gas in hollow cathode lamps and as sample
carrier in graphite furnace analysis in AAS.
A. Helium
B. Nitrogen
C. Argon
D. Acetylene

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 REFLECTION
I. Directions: Do the KIM (Key, Information, Memory). Complete the following grid
with ideas or pieces of information indicated by the headings.

Key Terms Information/Meaning Memory Clues (sentence

expressing your experience
about the key term)
Monochromator
Atomisation
Absorbance
Aspiration
Excitation

II. Self-Assessment: Answer the following questions on the right side of the circles, write your
answer on the circles.

What have you learned?

Why is this topic significant for you to learn as a

student researcher?

Can this topic help you in the future?

"The more that you read, the more things you will know. The more that you learn, the
more places you'll go." - Dr. Seuss

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