SUBJECT FORENSIC SCIENCE
Paper No. and Title Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods
Module No. and Title
in Forensic Science
Module Tag FSC_P4_M35
FORENSIC SCIENCE Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods in
Forensic Science
TABLE OF CONTENTS
1. Learning Outcomes
2. Introduction
3. Basic Principle of Instrumentations
4. Important Terminologies Associated with Instrumentation
5. Classification of Instrumental Techniques
6. Application of Instrumentation in Forensic Science
7. Summary
FORENSIC SCIENCE Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods in
Forensic Science
1. Learning Outcomes
After studying this module, you will be able to know
About the introduction of instrumentation
About principles of Instrumentation followed by the several instrumental methods.
Application of instrumental techniques in Forensic science
Advantages and limitations of instrumental techniques
2. Introduction
Analytical chemistry is a branch of pure chemistry which is very similar to physical chemistry.
The main objective of this branch of science is to develop and employ new methods and
instrumentation for the purpose of providing information on nature and composition of matter.
It helps in determination of a compound’s total or partial structure in various samples. The
word chemistry used in “analytical chemistry” clearly signifies the analysis of chemical
elements and their derived compounds. This branch of science is used in all the fields of
sciences.
Chemical sciences serves as proof of various advances and evolution of technologies which
has led to the growth of high performance instruments. This development of instrumentation
has led to increase in more sophisticated and non-destructive method of analysis. Non-
destructive techniques plays the most valuable role in forensic science as they can be conducted
on very small samples and does not even require extensive sample preparation before the
measurement. With these advances techniques quality and precision requirements can be met
efficiently. This is an important step in the official recognition of the quality of the laboratory.
The use of various instrumental techniques has become an important part of chemical analysis
in various fields of science i.e. pure and applied science. Only a single instrument cannot solve
an analytical issue, instead numerous instrumental methods are necessary for efficient analysis
to a maximum extent.
3. Basic Principle of Instrumentations
A physical property of the sample to be analysed is chosen in instrumental analysis for
determination of its chemical composition. Analysis of the sample can be of any type i.e.
biochemical, organic, inorganic, physical or analytical. Irrespective of the type of analysis, the
main purpose to be served is to extract information about the composition of the sample. This
FORENSIC SCIENCE Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods in
Forensic Science
is the reason why instrumental analysis is also known as quantitative analysis. The type of
instrumental technique used depends on the property of the sample measured.
The basic principle of an instrument employed for chemical analysis is that it can convert
chemical information retrieved from the sample to a form which can be easily observed and
understood. The instrument chosen for analysis thus helps in:
Generation of a signal
Transformation of a signal to one of a different nature (transducer)
Amplification of the signal.
Though all the steps are not combined in all the instruments.
4. Important Terminologies Associated with Instrumentation
Analytical technique – It is a vital scientific phenomenon which prove to be useful
for providing information regarding composition of a substance. For example, Ultra-
violet Spectrophotometry is an analytical technique.
Analytical method – It is a definite application of a method to analyse a sample. It is
also known as instrumental method. UV spectrophotometric analysis of a dye mixture
is an example of an analytical method.
5. Classification of Instrumental Techniques
Instrumental techniques can be classified on the basis of three areas:
Spectroscopy
Electrochemistry
Chromatography
Various instrumental techniques are:
5.1. SPECTROSCOPIC TECHNIQUES
5.1.1. Ultraviolet-Visible Spectroscopy
It is a type of absorption spectroscopy in ultraviolet-visible region of spectrum. This
spectroscopy uses light in the visible, near UV and near infrared ranges. The absorption in the
visible range directly affects the observed colors of the compounds involved. Atoms and
molecules undergo electronic transitions when exposed to radiations of this electromagnetic
spectrum.
FORENSIC SCIENCE Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods in
Forensic Science
5.1.2. Fluorescence and Phosphorescence Spectroscopy
Emission of photons from electronically excited states is known as luminescence. There are
two types of luminescence, fluorescence and phosphorescence. They both differ in the nature
of ground and excited state. After absorbing radiation from UV or visible region, the molecules
gets excited. This electronic excitation is supplemented with excitation of the molecule to
higher vibrational energy level. Due to loss if vibrational energy, the molecule return to the
ground state which leads to emission of light. This emission of light is known as fluorescence.
Phosphorescence is also emission of light when molecule return from excited state to ground
state of different spin multiplicity. The measurement of intensity of
fluorescence/phosphorescence as a function of wavelength is fluorescence and
phosphorescence spectroscopy.
5.1.3. Atomic Spectroscopy (Absorption and Emission)
The study of electromagnetic radiations which are absorbed or emitted by the atoms or
molecules is known as atomic spectroscopy. This spectroscopy is divided into atomic
absorption spectroscopy and atomic emission spectroscopy. Collection of atoms are exposed
to a particular wavelength of light. If the energy is in correspondence with the energy levels of
the atoms, a part of the light gets absorbed. The difference between the intensity of the light
emitted from the source and the light collected by the detector yields an absorbance value. This
absorbance value is then used to determine the concentration of the sample. The intensity of
emitted light is directly proportional to the concentration of the sample, in atomic emission
spectroscopy.
5.1.4. Infrared Spectroscopy
Infrared spectroscopy comprises of interaction of matter and infrared radiation. It is also known
as vibrational spectroscopy. It is also used to identify and analyse chemicals. Infrared
spectrometer is the main instrument used produce an infrared spectrum irrespective of the
nature of matter i.e. solid, liquid or gas. When the wavelength of IR radiation is identical to
vibrational frequency then absorption takes place. Similar to absorption spectroscopy, the
examination of transmitted light provides information about the amount of energy absorbed.
5.1.5. Raman spectroscopy
It is a spectroscopic technique which is used to observe rotational, vibrational and other low
mode frequencies. This spectroscopy is usually used to identify molecules. Its basic principle
is Raman scattering of monochromatic light.
5.1.6. X-ray spectroscopy
It is a commonly used method to determine atomic structure and electronic states. In this the
electron get excited and moved to unoccupied level or ejected from the atom.
FORENSIC SCIENCE Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods in
Forensic Science
5.2. ELECTROCHEMICAL TECHNIQUES
5.2.1. Potentiometry
It is one of the electrochemical technique which is used to find the concentration of a particular
solute in a solution. The potential between the two electrodes is measured while taking
potentiometric measurements. High impedance voltmeter is used in this.
5.2.2. Conductometry
In the presence of electric field ions in a solution move. Mobility of ions towards relevant
electrodes causes conductivity of ionic solutions. This measuring of conductivity of ionic
solutions is known as conductometry.
5.2.3 Voltammetric techniques
These techniques involve application of potential to an electrode and the resulting flow of
current, flowing through the electrochemical cell is observed.
5.3. CHROMATOGRAPHIC TECHNIQUES
5.3.1. Gas chromatography (GC)
It is a common type of chromatography which is used to separate and analyse compounds
which can be vaporized without getting decomposed. A carrier gas usually helium or nitrogen
is used as a mobile phase and a microscopic layer of liquid or polymer on an
inert solid support, inside a piece of glass or metal tubing called a column is used as stationary
phase.
5.3.2 Thin Layer chromatography (TLC) ·
It is a very convenient method. A thin even layer of the solid stationary is deposited on a glass
plate. The substance to be identified is processed as in paper chromatography. The results are
quicker and more accurate. The stationary phase and mobile phases can be varied at will. TLC
is a method of choice for most of the preliminary investigations, purification and identifications
of narcotics, drugs and poisons.
5.3.3. High Performance liquid chromatography
HPLC is a type of column chromatography in which a sample mixture or analyte in a solvent
is used as mobile phase which is pumped into the column (stationary phase) at high pressure.
The sample is carried using a carrier gas stream of nitrogen or helium. This technique separates
and identify various compounds present in the sample.
6. Application of Instrumentation in Forensic Science
FORENSIC SCIENCE Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods in
Forensic Science
Forensic analysis mainly comprise of identification of the samples i.e. qualitative determination
of identity of a substance on the basis of their physical and chemical properties. But only
qualitative analysis is not enough to give complete information about the sample and does not
complete the forensic investigation. Demonstration of common origin between two objects is
referred to as individualization.
Complete forensic analysis comprise of:
(1) Identification of the physical evidence
(2) Collection of the physical evidence following appropriate preservation and documentation
procedures
(3) Surrender of the evidence for analysis
(4) Examination of the evidence, which include identification and individualization processes
(5) Elucidation of the results of the examination
(6) Reconstruction of events according to the results obtained
Various types of physical evidences are encountered at the crime scene which may be physical,
chemical or biological. A simple classification scheme for such evidence is discussed in Table
1.
FORENSIC SCIENCE Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods in
Forensic Science
The emphasis here is on instrumental techniques of analysis which are most relevant to the
evidence and are classified under chemical or biological. Categorization of evidence according
to the applicability of the instrumental methods is listed in Table 2 whereas classification of
different types of instrumental methods and the type of physical evidence for which that
particular analytical technique is useful is discussed in Table 3.
FORENSIC SCIENCE Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods in
Forensic Science
More definitive results can be produces for various evidence/samples to analysed using
instrumental techniques. For example, identification of gunshot residue is possible using
various qualitative tests but elemental analysis techniques such as Atomic Absorption
Spectroscopy or Neutron Activation Analysis have proved to be appropriate for detection levels
of heavy metals present in gunshot residue analysis such as lead, barium, antimony etc. Various
other techniques like SEM, X-ray elemental analysis can also be used to visualize gunshot
residue particles and its elemental composition.
FORENSIC SCIENCE Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods in
Forensic Science
Various forensic analysis involve comparison of specimen and questioned sample. Several
instrumental techniques can be successfully used for comparison of samples. For example,
paint samples can be compared using Infrared Spectroscopy, two complex engine oils can be
compared using spectrophotofluorometry. Even for questioned documents various microscopic
techniques are used. Sequence of strokes in documents can be determined using
stereomicroscope and confocal microscope. In every case, either the sample do not match and
thus proved to be of different source or they match and are said to be consistent with the
specimen sample.
7. Advantages and Limitations of Instrumental Techniques
Advantages of Instrumental Analysis
The major advantages of instrumental analysis are:
Even minute quantities of samples are enough for the analysis
Reliable results are obtained.
It is easy to analyse complex samples as well
The rate of determination is fast and helps in complete individualization of the
sample.
Limitations of Instrumental Analysis. ·
There are various limitations of Instrumental analysis as well. These are:
The precision and sensitivity depend on the instrument and will vary accordingly.
The cost of the equipment is quite high. ·
The range of concentration that would be measured is limited. ·
Handling of every instrument is suggested only after training.
Appropriate space is needed.
There must be no fluctuation in the power supply.
8. Summary
The main objective of this branch of science is to develop and employ new methods
and instrumentation for the purpose of providing information on nature and
composition of matter.
This development of instrumentation has led to increase in more sophisticated and non-
destructive method of analysis.
FORENSIC SCIENCE Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods in
Forensic Science
The basic principle of an instrument employed for chemical analysis is that it can
convert chemical information retrieved from the sample to a form which can be easily
observed and understood.
Emission of photons from electronically excited states is known as luminescence.
The measurement of intensity of fluorescence/phosphorescence as a function of
wavelength is fluorescence and phosphorescence spectroscopy.
It is a common type of chromatography which is used to separate and analyse
compounds which can be vaporized without getting decomposed.
Forensic analysis mainly comprise of identification of the samples i.e. qualitative
determination of identity of a substance on the basis of their physical and chemical
properties.
More definitive results can be produces for various evidence/samples to analysed
using instrumental techniques.
Identification of gunshot residue is possible using various qualitative tests but elemental
analysis techniques such as Atomic Absorption Spectroscopy or Neutron Activation
Analysis have proved to be appropriate for detection levels of heavy metals present in
gunshot residue analysis such as lead, barium, antimony etc. \
Paint samples can be compared using Infrared Spectroscopy, two complex engine oils
can be compared using spectrophotofluorometry.
Sequence of strokes in documents can be determined using stereomicroscope and
confocal microscope.
In every case, either the sample do not match and thus proved to be of different source
or they match and are said to be consistent with the specimen sample.
One of the advantage of instrumentation is reliable results which can be obtained for
complex samples as well.
The precision and sensitivity depend on the instrument and will vary accordingly.
The cost of the equipment is quite high. ·
FORENSIC SCIENCE Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods in
Forensic Science
The range of concentration that would be measured is limited. ·
FORENSIC SCIENCE Paper No. 4: Instrumental Methods & Analysis
Module No. 35: Significance of Instrumental Methods in
Forensic Science
