REVIEW ARTICLE Am. J. PharmTech Res.
2020; 10(02) ISSN: 2249-3387
Journal home page: http://www.ajptr.com/
Review On Recent Analytical Techniques Used in Forensic Sciences
Mohan Raj KV1*, Vinod Kumar K1, Prashant D Ghode2, Bhargav E1
1.Department of Pharmaceutical Analysis, Raghavendra Institute of Pharmaceutical Education
and Research, KR Palli cross, Anantapuramu-515721
2.Department of Pharmaceutical Analysis, JSPMs Rajarshi Shahu College of Pharmacy and
Research, Tathawade, Pune-411033
ABSTRACT
The intention of this paper was to review and discuss the some of the quantitative analytical methods
used in forensic science for the analysis of trace evidences collected from the various scene incidents.
The forensic scientists were depend upon the instrumental analysis of trace amounts of materials
like drugs, toxicological specimens, fibers, glass, GSR, soil, etc. Differences in manufacture of
chemical composition are observed and that allows considerable discrimination even with very small
traces of fragments. Most on GSR, GSR analysis sometimes becomes very important to right
direction of inquiry, from where the GSR is obtained and from which surfaces the GSR has
produced, played an important aspect of investigations. Reviews on these techniques, which are used
more extensively in forensic sciences, were reported through this paper. Based on the analytical
problem facing forensic scientists, our study will summarize and use these techniques to tackle them
such as chromatography, Raman spectroscopy, Atomic Absorption Spectroscopy, Inductively
Coupled Plasma (ICP) techniques.
Keywords: Forensics, Chromatography, Atomic Absorption Spectroscopy, Tandem mass
spectroscopy.
Received 12 March 2020, Accepted 27 March 2020
Please cite this article as: Mohan KV et al., Review On Recent Analytical Techniques Used in Forensic
Sciences . American Journal of PharmTech Research 2020.
Mohan et. al., Am. J. PharmTech Res. 2020; 10(02) ISSN: 2249-3387
INTRODUCTION
Analysis of a crime scene involves expert participation in both the physical and biological sciences,
as well as in many technology and law enforcement areas. Analysis of a crime scene requires
specialist expertise both in the physical and biological sciences, as well as in other fields of
technology and law enforcement. Forensic scientists studies of fingerprint patterns and traces of
fiber, glass, bullet and other types, evaluate medicines and poisons, investigate suspected forgeries,
examine residues for potential incendiary and explosive crimes [1].
Forensic pharmacy means application of the drug sciences to legal issues like criminal cases [2].
Body fluids such as blood, seminal fluid, urine or saliva are very important objects in investigation
of crimes against someone or person such as murder and rape cases [3].
This review article reflects the theory and recent developments of modern technology for the study
of toxicants in biological fluids and tissues. We discuss limitations and innovations in methodology
that address these limitations. To fully understand instrument performance, one has to be familiar
with general analytical assay characteristics. These include the signal to noise ratio (S/N), limit of
detection (LOD), Limit of Quantification (LOQ), accuracy, precision, interference, and robustness.
Because of the importance of these parameters for characterizing assays and these are essential
components of method validation [4].
Many analytical instruments transform a certain analyte property into an electronic or photometric
signal. Several variables, such as random electronic or erroneous photon transmissions, fluctuating
concentrations of inherent substances in the detector and others contribute unwanted signals that can
interfere with measured target response. These aberrant signals are known as noise. The best
technologies improve analyte signal or reduce noise, or both, to increase S/N. High S/N results in
lower LOD and LOQ. LOD and LOQ are often defined as S/N = 3 and S/N = 10, respectively. These
limits are more generally calculated in forensic toxicology by measuring serial dilutions of an analyte
in the matrix of interest and choosing the lowest concentration where measurements satisfy S/N and
additional criteria [5].
In this paper, we have tried to explain the available analytical instrumental techniques available for
analysis of trace evidence analysis [1].
Toxicological chemistry examination using different chromatographic methods:
In almost all toxicology experimental studies, an agent, usually a single chemical, is given to an
organism in known amounts. It is widely agreed that the chemical under study must be pure, or that
the existence of any contaminants must be known to interpret the experimental results with validity
[6].
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However, it is common practice to conduct the instrumental study without verifying the compounds
purity. A scheme of separation for poisons from tissues by steam distillation and differential solvent
extraction is shown in Figure I
Figure I: Flow chart for Separation of Poisons from Tissues
The analytical techniques are initiated by forensic scientists have continued to enlarge in complexity
and improve in reliability. Several new analytical tools have proven useful for toxicology problems
in approximately all areas and the technology continue to explore open areas of research [1, 7]. The
x-ray fluorescence (XRF)/ X-ray diffraction (XRD) techniques have been used in forensic science
for several decades [1, 8]. Many other different analytical techniques, XRF does not affect samples
so it is called as a non-destructive method.
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This alone can be tremendously valuable as certain specimens involve confirmatory analysis by
another group of scientists to authenticate integrity of the data. Forensic science involves the
application of knowledge to law [9].
Forensic pharmacy is not an exotic field. Many issues relating to society are relevant to forensic
pharmacy including the physical traces like glass fragments, fibers, soils, building material, loose
powder materials etc. and also substance abuse control, impaired driving, crime, fraud, and
employment testing for any type of drugs. Pharmacists may provide valuable testimony in drunk and
driving cases, murder, suicide, malpractice, child abuse, personal injury and patents [2].
Possible applications of the XRF/XRD technique on different kind of forensic samples are described
in Figure II.
Elemental Chemical Analysis using XRD/ XRF
X-rays fluorescence /XRD in forensic examinations
Elements detection Purity check in Indian ornaments
Comparative examinations
1. Building Materials 1. Gold based ornament purity
1. Metals alloys
2. Soil 2. Silver based ornament purity
2. Paint chip
3. Drugs 3. Platinum based jewelry
3. Paper
4. Minerals
4. Automobiles under seals
5. Soap powder
5. Glass fragments
Figure II: Use of X – ray techniques in forensic sciences
Inductively coupled plasma (ICP) mass spectrometry technique
Mass spectrometry (MS) is an analytical technique that ionizes chemical compounds and detecting
the ions on the basis of their mass-to-charge ratio. Inductively coupled plasma mass spectrometry
(ICP-MS) is a type of mass spectrometry that can detect metals and several non-metals at
concentrations as low as parts per billion on uninterfered low background isotopes [10].
Discrimination of sheet glass exposed to high temperatures when trace impurities were determined
using ICP-MS was reported. Several reports on LIBS glass processing, and a comparison of LA-
ICPMS, µXRF and LIBS tests for sample analysis [11]. The determination of iron in glass by laser
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absorption and solution sampling comparing dynamic reaction cell-ICPMS and high resolution
ICPMS was also reported [1].
Emphasis on GSR analysis
Few knowledgeable experts all over the world commence the mission of documentation of the type
of bullets are taken into an account and identify the qualitative differences in chemical composition
of GSR (personal communication R Keeley, London Metropolitan Laboratory, Forensic Science
Service, 1997). Attempts an independent systematic approach to that investigation as
methodological study of gunshot residues originating from selected type of bullets by means of
SEM-EDX and uses the chemical methods for interpretation of the obtained results were obtainable
by Niewoehner [12] and Broiek-Mucha et al. [13, 14]. In their major presentation was depend on
quantitative analysis of out of hundred particles they are randomly taken from all of the preserved
gunshot residues from different components of bullet (i.e. including the primer, case, projectile
jacket etc.), whereas in the most recent two journals frequencies of incidence of primer particles of
particular chemical classes are taken into an account and experimental study were performed. By
knowing those both attempts results revealed the possibility of group identification of the bullets
used based on GSR study.
More sensitive than SEM-EDX, analytical methods are being introduced, and all are being used to
study the content of primary residues such as Inductively Coupled Plasma Mass Spectrometry (ICP-
MS)[15], provided more discriminative data. However the GSR analytical method only SEM-EDX
is the more accurate, given that information on both their morphology and the key critical contents
remains a reliable tool for GSR examinations [16].
Gas Chromatograph–Quadrupole Mass Spectrometer (GC–QMS) and GC– MS–MS
The GC–QMS is also called as GC–mass selective detector or GC–MS, commonly that requires the
analytes are chemically extracted from blood, urine, or other matrices and, in most cases,
derivatization technique is used to make them volatile and prior to introduction into the instrument
[4]. The GC separates the compounds based on differences in solubility and volatility in the liquid,
solid, and gaseous phases [5].
Molecules enter into the ion source sequentially, they are ionized. Electron ionization (EI) is the
most common ionization technique. Molecules leaving the GC enter the QMS and are bombarded
by a beam of electrons. Electrons are removed from the molecules, producing unstable positive ions
(molecular ions) which break into more stable fragments.
Chemical ionization (CI) is a “soft ionization” technique utilizes a charged reagent gas (NH4 or
CH4) to transfer charge to a compound. These charged species are more stable than the EI ions and
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fragments are formed less extensively. Both positive and negatively charged molecules are formed.
The QMS will select ions and measure their abundance from different mass-to-charge (m/z) ratios.
Specific ions are isolated by creating a dynamic electromagnetic field inside the quadrupole [4]. The
QMS can be configured to scan all the mass-to-charge ratios in its mass range and track specific
mass-to-charge ratios.
In forensic toxicology laboratories, GC–MS–MS procedures are sufficiently more common
following advances in vacuum engineering, ion sources, instrument size and operating software.
GC–MS–MS procedures were improved for measuring low concentration of analytes (e.g., LSD) in
blood or urine [17]. Current GC–MS–MS methods improve S / N in essential experiments with
LODs of 0.1 ppb (5 pg on column) including examination of metabolites of the nerve agent and
sulfur mustard in urine and metabolites in alternative matrices [18- 20].
Raman Spectroscopy Techniques
Raman spectroscopy is one technique used in forensic sciences. This method allows the
measurement of inelastic light dispersion due to the molecular vibration modes when irradiated by
a monochromatic source, such as a laser. The Raman technique has many benefits, such as its non-
destructive nature, its rapid analytical time and the possibility of performing microscopic in situ
analysis. This is a multi-purpose technique in its forensic application that covers a broad variety of
samples, such as drugs of abuse, physical / trace evidence[21], fibers and inks. The analysis of textile
fibers constitutes a clear example. In this field a number of studies have been carried out (e.g.
Bouffard et al. [22]; Keen et al. [23]. The value of this technique in forensic analysis of fibers mainly
focuses on the detection of dyes.
Atomic Absorption Spectroscopy
Hair analysis is done by Atomic Absorption Spectroscopy after sodium hydroxide and hydride
generation technique. Many of the arsenic particles on the hair have also been measured in KEK-
PF’s BL-4A (Photon Factory at High Energy Accelerator Research Organization, a synchrotron
radiation facility in Tsukuba) and found an arsenic particle on the hair. This method would allow the
measurement of inelastic light dispersion due to molecular vibration modes when irradiated by a
monochromatic source such as a laser. The size of the beam of synchrotron radiation along with the
hair shaft was 4 or 1 mm in diameter. How many hair shafts were counted, and how many particles
were found, is still not clear. It is said that arsenic particle was found on only one or two hair shaft
out off hundreds of hair shafts [24,25].
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LC tandem Mass Analysis
The tandem Mass spectrometer [26] is generally used an electrospray ionization source in a positive
mode. The chromatograms are separated by using suitable columns. At room temperature, mobile
phase Solvent A consisted of 5% acetonitrile with 0.1% formic acid and Solvent B consisted of 95%
acetonitrile with 0.1% formic acid. The chromatography system is operated at gradient mode by
changing the solvent ratios at different time intervals.
The urine samples are generally stored at 200C until analysis. Take 1mL urine sample and add 0.5mL
1.5M sodium bicarbonate buffer (pH 9.5) and 3mL ethyl acetate for liquid-liquid extraction. The
samples are mixed on a suspension mixer, centrifuge for 3 minutes and collect the supernatants were
decanted, dried under nitrogen gas. The residues were dissolved in 0.5mL of 5% acetonitrile with
0.1% formic acid. The mixture was vortex for 10 seconds and filtered through a 0.22µm
polyvinylidenedifluoride filter into a small volume of auto sampler vial. Then, 50µL was injected
into the LC-MS/MS system. All samples are prepared in this same manner [27].
CONCLUSION:
The analytical techniques are initiated by forensic experts and that have been continued to expand
the complexity and improves the reliability. Many analytical techniques are used to evaluate
analytical problems in nearly all fields of research and the technology continues to open areas in
every field of research. Forensic examiners are continued to determine the toxic substances and drug
abuses by different tools. Atomic Absorption Spectroscopy is mostly used technique to determine
the metallic compounds. The problems of drug abuse, increased therapeutic potency of drug
substances and widespread concern about the pollution and the safety and health of workers are the
present challenges to the analyst’s skills. Today investigators are uses a wide range of analytical
tests and sophisticated equipment with which to study the microscopic characteristics which are
collected at crime scenes.
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