UNIT 2 PHYSICAL INSTRUMENTATION IN

ENVIRONMENTAL SCIENCES

CONTENTS

1.0 Introduction
2.0 Objectives
3.0 Main Content
3.1 Instrumental Techniques for Heavy Metals Analysis
3.1.1 Atomic Absorption Spectrometry (AAS)
3.1.2 Flame Atomic Absorption Spectroscopy (FAAS)
3.1.3 Graphite Furnace Atomic Absorption Spectroscopy
(GFAAS)
3.2 Neutron Activation Analysis (NAA)
3.3 Instrumental Techniques for Organic Residues Analysis
3.4 The use of Gas Chromatography (GC) in Analysis
3.5 High – Performance Liquid Chromatography (HPLC)
4.0 Conclusion
5.0 Summary
6.0 Tutor-Marked Assignment
7.0 References/Further Reading

1.0 INTRODUCTION

In environmental sciences, all the pollutants of interest can readily be

classified as organic or inorganic. These ones are of greater concern
because they occur more often than any other ones we may think of.

Inorganic pollution arises from mining and smelting of metals, fossil

fuel combustion and chemical production coupled with widespread
applications in engineering, electronics, industrial and agricultural
practices. These activities have led to the presence of heavy metals and
other trace inorganic chemicals in the atmosphere, rainfall, rivers,
groundwater, soil, sediments and the biota. Organic pollution was first
manifested following the growth in the use of pesticides in the years
immediately after the Second World War and through the 1950s. The
first organochlorine pesticides were DDT, Lindane and Dieldrin.
Overuse and misuse of these compounds led to the death of wildlife,
especially species at the apex of food chains including raptorial birds,
foxes and badgers.

Public concern requires that pollutants in the environment are detected

and controlled. Current methods popularly used in profiling heavy metal
levels in an environmental matrix include Flame Atomic Absorption
Spectrometry (FAAS), Graphite Furnace Atomic Absorption
Spectrometry (GFAAS), Inductively Coupled Plasma-Mass
Spectrophotometry (ICP-MS) and Energy Dispersive X-Ray
Fluorescence (EDXRF) to mention a few. Chromatography is the
dominant analytical technique for the identification and quantification of
organic pollutants.

2.0 OBJECTIVES

At the end of this unit, you should be able to:

list mostly used instrumental techniques for the analysis of heavy

metals and organic compounds in an environmental sample
state the principle of operation of each instrumental technique
studied
state the advantages of one technique over the other.

3.0 MAIN CONTENT

3.1 Instrumental Techniques for Heavy Metals Analysis

Various, instrumental techniques are used by chemists and other

environmental scientists for the purpose of detecting and determining
the levels of heavy metals in a given environmental samples. For the
sake of brevity, only two of the techniques (AAS and NAA) are
discussed here.

3.1.1 Atomic Absorption Spectrometry (AAS)

Atomic Absorption Spectrometry is a technique that involves the

aspiration of the sample solution into a flame or an electrothermal
device whose high temperature converts the analyte ions into atoms in
the vapour state. When an electromagnetic radiation characteristic of the
electronic transitions of atoms of a particular element is passed through
an atomic vapour of that element, the radiation at certain frequencies is
attenuated. The absorbed radiation excites electrons from the ground
state to various higher energy levels (excited states). The degree of
absorption is a quantitative measure of the concentration of ground-state
atoms in the vapour.

AAS is the most widely used techniques for the quantitative

determination of metals at trace levels (0.1 to 100ppm) in a wide range
of materials; its relative precision is 0.5 to 2percent.

The major disadvantages include: (i) samples must be in solution or at

least volatile; (ii) individual source lamps are required for each element;
(iii) the technique is not capable of simultaneous multi-elemental
determination; and (iv) it is not suitable for qualitative analysis.
Some of the various modifications of AAS are Flame Atomic
Absorption Spectroscopy (FAAS) and Graphite Furnace Atomic
Absorption spectroscopy (GFAAS).

3.1.2 Flame Atomic Absorption Spectroscopy (FAAS)

FAAS consists of a sharp-line radiation source, produced by a hollow-

cathode lamp, characteristic of the element of interest, a solution
nebulizer and burner, a monochromator, photomultiplier and recording
system. Although FAAS is simple to operate and cheap, the burner-
nebuliser system is relatively an inefficient sampling device. Only a
small fraction of the sample reaches the flame and the atomised sample
passes quickly through the light path thereby leading to a low detection
limits, usually at the sub g/g or g/mL levels. Its dynamic range is
also limited.

Oxidant Fuel
Optical Path

Burner Head

Oxidant Excess Sample Spray Chamber

Aspirator
Sample

Fig. 6: The Premix Burner of a FAAS

Source: D. Harvey, 2000 (Modified).

3.1.3 Graphite Furnace Atomic Absorption spectroscopy

(GFAAS)

One major instrumental difference between GFAAS and FAAS is that,

graphite tube furnace (about 5 cm x 3 mm) is used in GFAAS in place of
flame in the FAAS for the purpose of vapourisation and atomisation.
The graphite tube furnace is flushed through with an inert gas, e.g.
argon, before vapourising the sample so as to prevent the formation of
refractory oxides and oxidation of the graphite tube. The axis of the
furnace is aligned along the optical path of the radiation from the lamp.
The sample (5 to 50 L) is deposited on the platform at the bottom
inner surface of the tube near the centre to enhance maximum
sensitivity. The temperature is rapidly raised to about 2500 K by the
 passage of a heavy current for a period of 1 to 2 minutes. The heating
cycle is controlled so as to allow solvents to evaporate or organic
residues to be ashed before an atomic vapour of the metal under
investigation is produced.

GFAAS has relatively low detection limits capability, which makes it

particularly suited to the requirements of analyses of trace elements at
low concentrations in a matrix. The main drawbacks of GFAAS are: (i)
it is not multi-elemental; (ii) it has limited practical sample throughout;
and (iii) the presence of high electrolyte species such as Na and Cl
results in numerous non-specific absorption interferences.

Hole
Graphite tube

Detector Output signal Recorder

Source

Furnace

Fig. 7: A Block Diagram of GFAAS

3.2 Neutron Activation Analysis (NAA)

NAA is a non-destructive tool for routine trace element determination in

many areas of innovative research. Apart from being able to determine
many environmentally crucial trace elements such as Sb, Cd, Cr, Cu, Se,
Ni, Zn, etc. NAA is also capable of determining major elements such as
Na, Cl, and K as well as rare earth elements.

The determination of the elemental concentrations is based on the

measurement of induced radioactivity through the activation of the
elements by neutrons. The radioactive decay of each element emits a
characteristics X-ray spectrum. Hence, an individual nuclear
“fingerprint” can be measured and quantified. The neutron sources used
in NAA can be produced by a neutron reactor, a particle accelerator or
artificial isotopes such as phitonium, and beryllium. The most common
source is from a fission reactor due to its high neutron flux.

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NAA is an extremely sensitive, selective and precise technique that
provides both qualitative and quantitative information at ultra trace
levels. These characteristics derive from a combination of factors: (i)
Extremely sensitive instrumentation with a facility for spectrometric
distinction between radionuclide is available; (ii) activation cross
sections can be large and interise neutron fluxes are available; (iii) the
reagent blank problem which is so common in trace element analysis is
largely eliminated; (iv) when sample processing prior to measurement is
needed, the problem of working with g amounts of materials can be
simplified by the addition of non-active “carrier’ which does not affect
the final activity measurement; and (v) very small sample size (flakes of
paint, single hair strand, etc) can be analysed and identified by NAA.

Some of the disadvantages of NAA are; (i) liquid samples cannot be

activated in a standard thermal neutron nuclear reactor; (ii) practical
multi-elemental analysis is restricted due to the wide spectrum of short
and long-lived nuclides; (iii) for short-lived nuclides measurement, the
presence of major electrolyte species (Na, Cl, P, Br) on irradiation
produces high background x-ray activities; (iv) the determination of
many important elements like Be, B, Pb, P and Si is difficult due to poor
nuclear cross-sections not activated by neutrons or, as in the case of P,
not yielding a X-ray for analysis; and (v) it is an expensive and highly
specialized instrumentation.

Activity: Read about and prepare “contact period” or term papers on

EDXRF and ICP-MS.

SELF ASSESSMENT EXERCISE 1

1. List some instrumental techniques suitable for the analysis of

heavy/trace metals
2. What are the advantages of (a) FAAS over GFAAS; (b) GFAAS
over FAAS; (c) AAS over NAA; and (d) NAA over AAS?
3. Explain the need of an inert gas to flush through the graphite
furnace tube before vaporisation of an element is commenced in
GFAAS analysis.

3.3 Instrumental Techniques for Organic Residues Analysis

In environmental chemistry, chromatography is the dominating

technique with respect to organic matrix analysis. Chromatography is
an instrumental analytical technique that combines separation and
identification of components of a complex mixture into individual
entities. The chromatographic methods have good speed, high resolution
power and tendency to handle small amounts of material.

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Every chromatographic system consists of moving or mobile phase in
intimate contact with fixed or stationary phase. The latter is composed
of the stationary or all non-moving portion of chromatographic column
or bed. A sample component undergoes an equilibrium distribution
between these two phases. This equilibrium in turn decides the velocity
with which each component migrates on column. The band-broadening
and dispersion of each component in the direction of migration also
occurs. Thus differential migration and band-broadening decide the
extent of separation of the sample.

The classes of chromatography are:

(i) Adsorption chromatography: This method is based on

exploitation of the difference in adsorptivity of solute to the
stationary support which is usually packed in a column e.g.
various fatty acids can be separated by adsorption
chromatography.
(ii) Partition chromatography: includes chromatographic
techniques such as liquid-liquid chromatography (LLC), paper
chromatography (PC) thin-layer chromatography (TLC), gas-
liquid chromatography (GLC) and reversed – phase
chromatography (RPC). Here, we explore the difference in the
partition coefficient or distribution ratio of individual species
in the mobile and stationary phase. Some partition
chromatography techniques like PC and TLC use plates while
others use columns.
(iii) Ion exchange chromatography: This method is based on
differences in the exchange potential between various ion
exchange resin packed in a column. Examples are CEC, AEC,
IE and Liquid Exchanger (LE).
(iv) Exclusion chromatography: This is based fundamentally
upon exploitation of the difference in size or molecular
geometry of the components. In gel permeation, small
constituents are retained in inter shell spaces or pores while
large size components emerge first. Examples include gel
permeation (GP), ion exclusion and molecular sieve
chromatography.
(v) Electro Chromatography: Methods under this category are
often classified as electrophoretic techniques. In such
separations, the difference in mobility of different ions when
an external potential is applied. Examples include zone
electrophoresis, boundary layer electrophoresis, curtain
chromatography and capillary electrophoresis of all the
chromatographic techniques available, GC and HPLC are
used more often for routine analysis of environmental
samples.

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3.3.1 The Use of Gas Chromatography (GC) in Analysis

With GC, it is possible to separate a very complex mixture containing up

to 200 or more related compounds, using either partition or adsorption,
with very small sample sizes. It is similar to liquid-liquid
chromatography except that the mobile liquid phase is replaced by a
moving gas phase. The stationary phase may be solid or liquid.

Flow Meter
Injector Septum
block Detector
Amplifier
Pressure
control
Oven Data Output

Column Chart trace

Fig. 8: Diagram of a Gas Chromatograph

Source: D. Harvey, 2000 (Modified).

The sample for GC analysis must be able to exist in the gas phase, so it
may be applied to volatile materials only. Thus, non-polar substances
are easier to handle than polar materials; ionic materials cannot pass
through a GC. For polar substances like alcohols, amines, free fatty
acids and phenols, derivatisation may be required.

Ph – OH + Cl - SiMe3 Ph – O – SiMez + HCl

Phenol Trimethylsilyl Phenyltrimethylsily
chloride ether

The carrier gas, from a high-pressure cylinder is helium, nitrogen,

hydrogen or argon. The choice depends on factors such as availability,
purity, consumption and type of detector.

Until recently, GC analysis was handled using packed columns in which

the stationary phase is a liquid that has been coated on an inert granular
solid called the column packing (held in borosilicate glass tubing).
More recently, however, the borosilicate glass tubing packed columns
are being replaced by fused silica or quartz capillary columns. The
column is installed in an oven with the inlet attached to a heated injector
block and the outlet attached to a detector. Precise and constant

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temperature control of the injector block, oven and detector is
maintained. Stationary phase material and concentration, column length
and diameter, oven temperature, carrier gas flows and detector type are
the control variables.

Sample solution is usually introduced using a microsyringe with

hypodermic needle inserted through a self-sealing silicone rubber
septum. The sample is smoothing injected into a heated metal block at
the head of the column. Modes of placing samples onto the column can
be by split injection or splitless injection. Manipulation of the syringe
is an art developed with practice, and the aim is to introduce the sample
in a reproducible manner. The temperature of the sample port should be
such that the liquid is rapidly vaporised without either decomposing or
fractionating the sample. A useful rule of the thumb is to set the sample
port temperature approximately to the boiling point of the least volatile
component. For greatest efficiency, the smallest possible sample size (1-
10 L) consistent with detector sensitivity should be used.

In GC sample analysis, interferences may arise from contamination of

samples, chromatograph improper functioning and countermeasures that
may manifest inform of septum bleed, column bleed and ghost peaks
manifestation.

Gas Chromatograph Detectors

The function of a GC detector is to sense and measure the small amounts

of the separated components present in the carrier gas leaving the
column. The choice of a detector will depend on factors such as the
concentration level to be measured, the nature of the separated
components and the properties of the detector e.g. high sensitivity, good
linearity, stability and response.

a) Hot-wire director (HWD): This is also known as the thermal

conductivity detector (TCD) or katherometer. It is the oldest
GC detector. Due to its inherently large volume, low sensitivity
and contamination problems, it was long dismissed as unsuitable
for capillary systems. It is universal in its applications.
b) Electron capture detector (ECD): This usually used for the
analysis of compounds that have high electron affinities such as
chlorinated pesticides, drugs and their metabolites. This detector
is somewhat selective in its response, being highly sensitive
towards molecules containing electronegative groups: halogens,
perozisdes, quinines and intro groups. It is insensitive towards
such functional groups as amines, alcohols, and hydrocarbon
c) Flame ionization detector (FID): This more or less universal
detector is widely used because of its high sensitivity to organic

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 carbon-containing compounds. It is perhaps the most widely used
detector for GC. Its advantages include: (i) it responds to virtually
all organic compounds with high level of resolution; (ii) it is
resistant to common carrier gas impurities such as resistant to
common carrier gas impurities such as water and carbon; (iii) it
has a large linear responds range and excellent baseline stability;
(iv) it is relatively insensitive to small column flow-rate
fluctuations during temperature programming; (v) it is highly
reliable, rugged and easy to use; and (vi) it has low detector dead-
volume effects and fast response. Its two major limitations are: (i)
it gives little or no response to non-combustible gases and all
nobles gases; and (ii) it is a destructive detector that changes both
the physical and chemical properties of samples analyzed
irreversibly.
d) Photoionization detector (PID): Photoionization occurs when a
molecular species dissociates into a parent ion and an electron
upon interaction with UV light.

M uv M+ + e-

The PID detects organic and some inorganic species in the

effluent of a gas chromatograph with a detection limit as low as
the pictogram range. The PID has a high sensitivity, low noise
and an excellent linearity. It is non-destructive and can be used in
series with a second detector for more selective detection. PID
can be operated as a universal or selective detector by simply
manipulating the photon energy of the ionization source.
e) Mass spectrometer (MS): Mass spectrometers can serve as
detectors when coupled to a GC. The MS combines the ability to
detect a wide variety of compounds with the capability of
deducing compound structures from fragmentation patterns or
mass spectra. The computer (for recording) contains and can
search a library of known mass spectra to identify tentatively an
unknown compounds are used for confirmation after tentative
identifications are made.
f) Fourier Transform Infrared Spectrometers (FT-IR): Like the
MS, FT-IR is also an independent instrument which can be
coupled to a GC to serve as a detector.
g) Thermionic detector: This particularly responds to compounds
containing nitrogen or phosphorus

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3.4 High-Performance Liquid Chromatography (HPLC)

Although GC is widely used, it is limited to samples that are thermally

stable and easily volatilised. Non-volatile samples, such as peptides and
carbohydrates, can be analysed by GC, but only after they have been
made more volatile by a suitable chemical derivatisation. For this
reason, the various techniques included within the general scope of
liquid chromatography are among the most commonly used separation
techniques.

In HPLC, a liquid sample, or a solid sample dissolved in a suitable

solvent, is carried through a chromatographic column by a liquid mobile
phase. Separation is determined by solute/stationary-phase interactions,
including liquid-solid adsorption, liquid-liquid partitioning, ion
exchange and size exclusion, and by solute/mobile-phase interactions. In
each case, however, the basic instrumentation is essentially the same. An
HPLC typically included two columns: the guard column and an
analytical column. The guard column is an inexpensive column placed
before the analytical column (a more expensive column), protecting it
from contamination and damage, while the analytical column does the
separation. In HPLC, the stationary phase is a liquid film coated on a
packing material consisting of 3-10 m porous silica particles.

Solvent proportioning valve

Solvent proportioning valve

Pump Pulse
damper

Guard

Column

Detecto Record

Fig. 9: Diagram of a High-performance Liquid

Chromatograph
Source: D. Harvey, 2000 (modified).

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The stationary phase may be partially soluble in the mobile phase,
causing it to “bleed” from the column over time. To prevent this loss of
stationary, it is covalently bonded to the silica particles by reacting the
silica particles with an organochlorosilane (Si(CH3)2 RCl).

The elution order of solutes in HPLC is governed by polarity; the least

polar solute spends less time in the polar stationary phase and is the first
solute to elute from the column. Retention times in a normal-phase
separation are controlled by selecting the mobile phase, with a less polar
mobile phase leading to longer retention times. In a reverse-phase
separation, however, the order of elution is reversed.

As with GC, numerous detectors have been developed for use in

monitoring HPLC separations. To date, the most HPLC detectors are not
unique to the method, but are either stand-alone instruments or modified
versions of the same. The most popular ones are spectroscopic detectors
(e.g. UV /visible absorption and fluorescence) and electrochemical
detectors (such as amperometry, voltammetry, coulometry and
conductivity based detectors). A refractive index detector is sometimes
employed as a universal detector.

SELF ASSESSMENT EXERCISE 2

1. Explain the term “chemical derivatisation” as it applies to GC

analyses. Advance two reasons why it is necessary.
2. List five detectors that are used in GC analysis.
3. With reasons, state the detectors you would adopt in carrying out
a GC analysis of the following: Peptides, (b) Chlordane, (c)
Gaseous hydrocarbons, (d) Mercaptans, (e) Carbohydrates.
4. List three advantages and two disadvantages of flame ionization
detector (FID) as a GC detector.

4.0 CONCLUSION

Qualitative and quantitative identification of inorganic and organic

species in a given matrix have become easier, faster and more
interesting because of the availability of the state –of –art instrumental
techniques of modern days. Apart from their applications by
environmental chemistry, such instrumental techniques have found wide
acceptance and applications in clinical analysis, consumer goods
characterization and petroleum products analysis.

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5.0 SUMMARY

In this unit, you have learnt that:

Levels of heavy metals in a sample can be profiled by such

instrumental techniques as AAS (FAAS and GFAAS), NAA,
EDXRF and ICP-MS.
Chromatographic techniques are well suited for the analysis of
organic compounds in particular.

6.0 TUTOR-MARKED ASSIGNMENT

1. Briefly explain the following modes of sample injection in a GC

analysis: (a) Split injection, (b) Split less injection.
2. With respect to interferences in GC analysis, explain the
following: (a)Septum bleed, (b) Column bleed and (c) Ghost
peaks.

7.0 REFERENCES/FURTHER READING

Mendham, J.; Denney, R.; Barnes, J. & Thomas, M. (2004). VOGEL’S

Textbook of Quantitative Chemical Analysis (6th ed.). Delhi,
India: Pearson Education Ltd.

Harris, D. (1999). Quantitative Chemical Analysis (5th ed.). New York:

W.H. Freeman and Company.

Khopkar, S. (2008). Basic Concepts of Analytical Chemistry. New

Delhi: New Age International Publishers.

Braun, R. (2000). Introduction to Chemical Analysis. London: McGraw-

Hill Int. Editions.

Harvey, D. (2000). Modern Analytical Chemistry. USA: McGraw-Hill

Companies Inc.

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