ABOUT US
Malaysian Journal of Forensic Sciences, a peer-reviewed annual journal initiated by Department of Chemistry
Malaysia, Royal Malaysian Police and Universiti Sains Malaysia for the Forensic Science Society of Malaysia
in conjunction with the 2nd anniversary of the establishment of the Society in 2010. It is a platform for the
dissemination of information and research findings from various branches of Forensic Sciences. These include
criminalistics, narcotics, toxicology, DNA analysis, serology, anthropology, forensic psychology and other
forensic-related fields. This Open Assess online journal, managed and maintained by Universiti Sains Malaysia,
Kubang Kerian operating on an on-line submission and reviewing system welcomes manuscripts on all aspects
of forensic science related article from local and international researchers.
MALAYSIAN JOURNAL OF FORENSIC SCIENCES (MJOFS): MISSION STATEMENT
The mission of Malaysian Journal of Forensic Sciences (MJOFS), the annual journal initiated by Department of
Chemistry Malaysia, Royal Malaysian Police and Universiti Sains Malaysia is to serve as a platform for
disseminating observations, findings, and views from various branches of Forensic Sciences by publishing peerreviewed articles in a balanced scientific and objective manner.
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Zafarina Zainuddin
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Mohd Kamarul Zaman Ibrahim
CONTACT INFORMATION
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Malaysian Journal of Forensic Sciences
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Email: mjofseditor@gmail.com
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Perpustakaan Negara Malaysia – ISSN 2180-4710
i
Volume 1, Number 1, October 2010
Contents
A Message from the Editor-in-Chief
1
From Narcotics Case Files
2
Kee-Bian Chan, Rusyidah Abdul Rahim, Hock-Sing Ng, Khuzaida Abdul Raof
More from Our Recent Case Files
6
Kee-Bian Chan, Suhana Ismail, Muzaiyanah M. Kaprawi
Determination of Formaldehyde Contamination in Wiping Media Used in
Trace Pseudoephedrine Analysis
12
Ahmad Fahmi Lim Abdullah, Gordon M Miskelly
Forensic Light Sources for Detection of Biological Evidences in Crime
Scene Investigation: A Review
17
Wee-Chuen Lee, Bee-Ee Khoo
Development of Reagent Test Kit for the Enhancement of Shoeprints at
Crime Scene
28
Umi Kalthom Ahmad, Noorul Huda Abdul Jabar, Chong-Hooi Yew, Noor Azmi Yusoff
Skull-Photo Superimposition: A Remedy to the Problem of Unidentified
Dead in Malaysia
34
P. T. Jayaprakash, Bhupinder Singh, Ridzuan Abd Aziz Mohd Yusop, Hetty Susilawati Asmuni
A Review on Solid Phase Microextraction and Its Applications in Gunshot
Residue Analysis
42
Kah-Haw Chang, Ahmad Fahmi Lim Abdullah
Analysis of Gunshot Residue Deposited on Cotton Cloth Target at Close
Range Shooting Distances
48
Mohamed Izzharif A. Halim, Umi Kalthom Ahmad, Chong-Hooi Yew, Muhammad Koey
Abdullah
A Study on the Age Related Retention of Individual Characteristics in
Hand Writings and Signatures for Application during Forensic
Investigation
54
Nataraja Moorthy T, Mohamad Mahathir Amir Sultan, Kong-Yong Wong
Identifying Profile of Female Prisoners in Malaysian Prison Using Carlson
Psychological Survey
60
Nurul Hazrina Mazlan, Geshina Ayu Mat Saat, Affizal Ahmad
ii
Malaysian Journal of Forensic Sciences, 2010, Vol.1, No. 1
A Message from the Editor-in-Chief
Greetings and welcome to the inaugural issue of
Malaysian Journal of Forensic Sciences.
analysis of gunshot residue deposited on cotton
cloth target at close range shooting distances.
The inaugural edition of Malaysian Journal of
Forensic Sciences has been initiated by Department
of Chemistry Malaysia, Royal Malaysian Police
Force and Universiti Sains Malaysia for the
Forensic Science Society of Malaysia. It is an
online refereed journal maintained by Universiti
Sains Malaysia, Kubang Kerian which is devoted
to the publication of original investigations,
observations, and reviews in the various branches
of forensic sciences. These include criminalistics,
narcotics, toxicology, DNA analysis, serology,
anthropology, forensic psychology and other
forensic-related fields.
Other articles focus on the use of forensic light
source for biological evidence detection, the
development of skull-photo superimposition as an
identification technique, a study on the age related
retention of individual characteristics in hand
writings and signatures for application during
forensic investigation, the development of reagent
test kit for the enhancement of shoeprints at crime
scene, and identifying profile of female prisoners in
Malaysian prison using Carlson psychological
survey.
The journal aims to provide a platform for
disseminating observations, findings, and views
from various branches of forensic sciences. It
establishes and maintains a review of the literature
of research and practice in forensic science and
crime scene related areas. It also serves as a means
for critical reflection and analysis of different
approaches used in the court of law. Recent trends
of academic paper writing and publication reflect
the fusion of ideas from diversified disciplines and
research approaches. This is indeed refreshing and
helps creative endeavours as the development of
novel
ideas
depends
largely
on
such
multidisciplinary contribution with new way of
looking at the same things and interpreting findings.
We are happy to accept and publish traditional
academic articles in written form at the beginning
stage of this journal; we also specifically invite and
encourage submissions on the Web. We are
looking for submitters who want to participate fully
in our journal as reviewers who can help review,
critique, and comment on other article submitted by
your peers.
Most importantly, we hope to continuously
improve the core publishing process of the journal
to meet international journal publication standard
in the near future. With active participation and
contribution from all submitters and reviewers, this
is achievable within a short space of time.
Yew Chong Hooi
In this inaugural edition there is article focusing on
the unusual encounters from narcotics case files; a
review on solid phase microextraction and its
applications in gunshot residue analysis; a study on
formaldehyde contamination in wiping media used
in trace pseudoephedrine analysis, and a study on
Editor-in-Chief
E-mail: mjofseditor@gmail.com
31 October 2010
1
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
From Narcotics Case Files
Kee-Bian Chana, Rusyidah Abdul Rahima, Hock-Sing Ngb, Khuzaida Abdul Raofc
a
Narcotics Section, Department of Chemistry Malaysia, Petaling Jaya, Malaysia
b
Department of Chemistry Malaysia (Penang branch), Penang, Malaysia
c
Department of Chemistry Malaysia (Kedah branch), Kedah, Malaysia
ABSTRACT: This paper reported several unusual encounters over the years in the Narcotics Sections of
Department of Chemistry. The first case reported a court case that led to amendment of definition of opium
poppy and raw opium in the Dangerous Drugs Act. The second case reported our encounter with condom-liked
packets following a surgical operation of a man suffering from severe abdominal pain, suspected to be
controlled substances recovered from a body, which was subsequently identified as iguana eggs. The last case
reported our experience with black gummy substance suspected to be opium which was encountered twice in
our laboratories. These cases delineate the unusual side of the work with drug cases in our laboratories.
Keywords: narcotics, opium, controlled substance
Introduction
In Malaysia the Narcotics Section at Petaling Jaya
and branch laboratories examine all the drug cases
submitted by law enforcement agencies for
prosecutorial and investigative purposes. In this
paper, we relate several unusual encounters over
the years.
1. Papaver somniferrum L vs Papaver setigerum
DC
Raw opium smuggling was rampant in the 70’s and
80’s with most of this drug coming through the
border in the north of Peninsular Malaysia. Nearly
all cases were handled by the laboratory of
Department of Chemistry branch in Penang since
most of the arrests were made near the border and
in the northern states of Penang, Perlis and Kedah.
Raw opium was trafficked mostly in multiple 2kilogramme packages, wrapped with layers of rice
paper and plastic sheets. The presumptive weight
for trafficking in raw opium is one kilogramme
which carries the death penalty on conviction.
In 1984, an issue arose in the Kangar High Court
over a raw opium trafficking case when the chemist
of Department of Chemistry was asked whether he
had ascertained that the raw opium was derived
from the species Papaver somniferrum L. (PSL)
and not from Papaver setigerum D.C. (PSDC).
Note that at that time, the Malaysian law was
explicit that only raw opium from PSL was illicit as
the Malaysian Dangerous Drugs Act (1952) then
defined “opium poppy” as
“a plant of the species Papaver somniferrum
L or the species Papaver setigerum D.C. and
any plant from which morphine may be
produced”
and “raw opium” as
“the spontaneously coagulated juice
obtained
from the
plant
Papaver
somniferrum, L., which has not undergone
the process necessary to convert it to
medicinal opium, whatever its content of
morphine”.
Although it had been reported that PSDC was not
known to be a source of raw opium, whether licit or
illicit, the chemist in the eyes of the court could not
convincingly answer the question based on the
analytical tests carried out at that time [1, 2]. A
literature search revealed that Japanese scientists
had profiled the alkaloidal contents of PSL and
PSDC and had shown that they are actually distinct
[3]. Based on this literature finding, the Department
of Chemistry Malaysia informed the prosecutorial
authorities and the High Courts that the laboratory
could carry out an alkaloid profiling to determine
the source of the questioned raw opium. As a result,
all the raw opium evidence had to be profiled to
determine if it had come from PSL and all raw
opium cases awaiting trial at the High Courts had
to be returned to the laboratory for retesting. A
total of 100 raw opium samples from both the fresh
cases and the returned evidence were therefore
profiled on their codeine, morphine, thebaine,
papaverine and noscapine contents using a packed
column Gas Chromatography method with
amitriptyline as the internal standard [4, 5]. A brief
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
procedure and results of analysis of these raw
opium samples is as follows:
i)
ii)
iii)
iv)
v)
vi)
Equipment: Perkin Elmer F11 Gas
Chromatograph equipped with a FID and a
Hewlett Packard 3390A integrator.
Column: Glass column 2m × 2mm i.d.
containing a packing of 3% OV-1 on 100120 mesh Supelcoport.
Temp. programme: 230°C to 280°C at a rate
of 5°C/min. and held at the final temp. for 3
minutes.
Internal standard: Amitriptyline HCl
(Roche); morphine standard (MacFarlan
Smith), all other alkaloid reference standards
from Sigma.
Analysis: Representative samples (ca. 2 g)
were dried at 100 °C for 3 h. The dried
samples were ground to a fine powder using
a mortar and pestle. Duplicate samples (0.5
g) were each extracted with 25 mL of
methanol-chloroform
(1:1)
by
ultrasonication for 15 mins. One mL of this
solution was mixed with 1 mL of the internal
standard solution (0.60 mg/mL of
amitryptyline HCl). The five principal
alkaloids were quantitated simultaneously
against a standard solution containing a
mixture of the alkaloid reference standards.
Summary of Results:
Component
Moisture
Codeine
Morphine
Thebaine
Papaverine
Noscapine
Mean
27
2.5
11.5
3
0.44
6.2
Range
15 - 39
1.6 - 4.2
6.4 - 17.4
1.6 - 5.3
0 - 1.2
2.1 - 9.6
Alkaloid profile of the 100 raw opium samples
(w/w %, dry basis)
In September 1984, the definition of “opium poppy”
and “raw opium” were amended to obviate the
chemist’s task of profiling. This amendment via
Act A596 now reads:
“opium poppy” means any plant from which
morphine may be produced;
and
“raw opium” means the coagulated juice
obtained from any plant from which
morphine may be produced, whatever its
content of morphine and in whatever form
the coagulated juice is, but does not include
medicinal opium.
A recent work [6] provided further proof that the
chemists from the laboratory of Department of
Chemistry branch in Penang had not erred in their
contention that the raw opium had indeed been
derived from PSL.
2. The Night of the Iguana
In the middle of a night, a man in extreme distress
and with severe abdominal pain was rushed to the
hospital. A surgical operation was performed but
when he awoke from the operation, he found that
he had landed in serious trouble with the law - the
doctor had recovered three condom-like packets
from his gastrointestinal tract and alerted the police.
Two of the packets appeared to have ruptured while
the third one remained intact (Fig. 1). He was held
in custody while the three packets were sent to the
Department of Chemistry to ascertain if they
contained controlled substances.
Fig. 1: One of the 3 packets recovered from the
gastrointestinal tract of the man
Quick screening tests were negative for common
controlled substances encountered in our
laboratories. It was also observed that the surface
of the packets had background lines appeared to be
fine blood vessels. When the initial findings were
relayed to the investigating officer, the chemist was
informed that the man had related the source of his
misfortune:The afternoon previous to the incident, he had been
urged by a friend to swallow three immature iguana
eggs with a Guinness Stout® drink to improve his
sex life. For comparison purpose, the chemist from
our laboratory purchased some specimen of
immature iguana eggs (Fig. 2) from the same
vendor at Sentul market.
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Fig. 2: Two specimen immature iguana eggs purchased
from the market
Fig. 4: GC-MS chromatogram of opium-like substance
with the peak at RT=21.788 min
It was found that the three condom-like packets
physically resembled the specimen. Both consisted
of a tough elastic outer layer with a homogenous
dough-like mass inside. The eggs might have been
too tough to digest or caused a gastrointestinal
blockage.
3. I can’t believe it’s not opium!
In 2008, the Alor Setar branch laboratory of
Department of Chemistry received a case
comprising some black gummy substance
suspected to be opium. The substance had very
similar physical appearance and odour of prepared
opium (Fig. 3).
Fig. 3: The opium-like black sticky substance (2008)
To the great surprise of the analyst, all screening
tests were negative; TLC and GC-MS failed to
detect any opium alkaloids. Instead, the GC-MS
analysis showed a major peak (RT=21.788, Fig. 4)
which its mass spectrum matched aloe-emodin
from the mass spectral library (Fig. 5).
Fig. 5: Mass spectra of the peak eluted at 21.788 min
matched the library spectrum of aloe-emodin
Merck Index indicates that aloe-emodin (Molecular
structure as in Figure 4, right) occurs in the free
state and as a glycoside in Rheum (rhubarb), in
senna leaves and in various species of Aloe
(Liliaceae) [7]. It is freely soluble in hot alcohol, in
ether, in benzene with yellow colour, in ammonia
water and in sulfuric acid with crimson colour [7].
This compound has been reported to have
antitumor, genotoxicity, antioxidation, and
antibacterial effects [8, 9] and can be used as
laxative. The substance was suspected to be
derived from the sap of the Aloe plant; however,
this could not be confirmed due to the lack of an
authenticated sample and literature data.
About 30 years earlier, the Penang branch
laboratory of Department of Chemistry had
encountered a similar case in which several large
tins of an opium-like substance were seized from
an abandoned boat thought to have come from
Myanmar (Burma). Screening tests and TLC ruled
out the presence of opium (Note that GC-MS was
not available then). A hint to the identity of the
substance came from a note in the cabin which bore
the word “aloes”.
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Conclusion
5.
We report some of the rare encounters over the
years in our narcotics laboratories. These unusual
experiences undoubtedly make our life as chemists
more colourful besides performing routine
narcotics laboratory analysis.
6.
References
1.
2.
3.
4.
MJ de Faubert Maunder. (1975). Field and
laboratory tests for raw and prepared opium.
Bulletin on Narcotics. 27(1): 71-76.
HY Lim, SY Kwok. (1981). Differentiation
and comparison of raw, prepared and dross
opium. Bulletin on Narcotics. 33(1): pp.31-41.
Haruyo Asahina et al. (1957). Studies of
poppies and opium. Bulletin on Narcotics. 2033.
Dmytro Furmanec. (1974). Quantitative gas
chromatographic determination of the major
alkaloids in gum opium. Journal of
Chromatography. 89: 76-79.
7.
8.
9.
F Van Vendeloo et al. (1980). Fingerprint
analysis of illicit heroin samples by gas
chromatography. Pharmaceutisch Weekblad
Scientific Edition. 2(5): 129-136.
S Panicker et al. (2007). Quantitation of the
major alkaloids in opium from the Papaver
Setigerum DC. Microgram Journal. 5(1-4):
13-19.
The Merck Index, 13th Edition (2001), pp. 56.
H.Z. Lee, S.L. Hsu, M.C. Liu and C.H. Wu.
(2001). European Journal of Pharmacology.
431: 287–295.
Zaffaroni, M et al. (2003). High-performance
liquid chromatographic assay for the
determination of Aloe Emodin in mouse
plasma. Journal of Chromatography B. 796 (1):
113-119.
Additional information and reprint requests:
Chan Kee Bian
(E-mail: kbchan@kimia.gov.my)
Narcotics Section
Forensic Division
Chemistry Department of Malaysia
46661 Petaling Jaya, Selangor, Malaysia
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
More from Our Recent Case Files
Kee-Bian Chana, Maimonah Sulaimana, Suhana Ismaila, Muzaiyanah M. Kaprawia
a
Narcotics Section, Department of Chemistry Malaysia, Petaling Jaya, Malaysia
ABSTRACT: This paper further reports some of the recent interesting cases encountered in our drug laboratories. Six
cases relating to the analysis of unknown substances are presented in the first part. In two of these, we report our first
encounter with the plant material khat and “ma huang” (ephedra). In the second part we report a case involving
concealed packets recovered from the gastrointestinal tract of an Iranian body packer where we found a raw opium
drug packet among the other 99 packets of crystalline methamphetamine hydrochloride. These cases reflect the
significant challenges faced by our forensic drug chemists while at the same time they provide a welcome relief to the
tedium and monotony of the mundane daily work.
Key words: narcotics, controlled substance, methamphetamine, khat, cathinone
Introduction
The Narcotics Section of Department of Chemistry
Malaysia at Petaling Jaya and branch laboratories
examine all the drug cases submitted by law
enforcement agencies for prosecutorial and
investigative purposes. We received samples of all
kinds and some unusual encounters related to our
routine work were reported earlier. In this paper, we
further report some unknowns submitted to us for
characterization and a recent unusual case of opium
smuggling through the airport.
a) The Unknowns
Case 1 (2005)
Owing to the nature of the work and the samples
handled, our forensic drugs laboratories very rarely
handle work from the private sector. We report a
strange case here: - A middle-aged man presented
some white crystalline substance to our laboratory for
analysis. The wife, who had been in good health, had
suddenly died of a heart attack a few days earlier. A
few weeks before the incident, the couple had newly
employed a maid to help with the household chores.
While they had no problem with the maid and were
quite happy with her work, the husband could not help
thinking that there might be a connection between the
maid and his wife’s sudden death. He became more
suspicious when he found a small plastic packet of
white crystalline substance in the maid’s drawer.
We screened the substance using Marquis, sodium
nitroprusside and Janovsky tests. The results of these
tests were negative and we ruled out the presence of
opiates, methamphetamine and other amphetamine
type stimulants (ATS), or ketamine. The substance
dissolved readily in water giving an acidic solution –
indicating possibly an inorganic substance. Further
analysis and material characterization by X-Ray
Diffraction Analysis (XRD), Fourier Transform
Infrared Spectroscopy (FTIR) looking at its organic
materials and Scanning Electron Microscopy (SEM)
identified the substance to be alum, a relatively
harmless chemical and thus ruling it out as a smoking
gun.
Case 2 (2005)
The police seized a plastic packet of white crystalline
substance from a lady going through airport
immigration. She was detained for possession of the
substance suspected to be a controlled substance and a
sample was submitted to our laboratory for analysis.
We conducted various colour tests routinely used for
common controlled substances followed by FTIR and
SEM analysis. The substance was determined to be
sodium borate (borax).
Case 3 (2007)
An off-white powder was found strewn on the grassy
open ground in the vicinity of a methamphetamine
clandestine laboratory. The substance was submitted
to the laboratory for further determination. Initial
visual examination indicated the substance could have
been there and exposed to the elements for some time.
The substance was probably insoluble in water as it
had not been washed away during tropical rain. Our
intuition told us that it is unlikely to be
methamphetamine hydrochloride salt produced by the
clandestine laboratory or a methamphetamine-related
precursor such as pseudoephedrine or ephedrine
powder since they would be too valuable to be
discarded. Initial guess of the identity of this unknown
was either diatomaceaous earth or fuller's earth and it
6
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
had been utilized as a filtration or cleaning aid. The
SEM trace of this substance (Fig. 1) resembled that of
feldspar’s which was provided by UNODC (Vienna).
Note that Feldspar is the mineral name given to a
group of minerals distinguished by the presence of
aluminum (Al) and the silica ion (SiO4) in their
chemistry.
Fig.1: SEM trace of the off-white unknown
Case 4 (2010)
A consignment of fresh plant material seized by the
customs was submitted to our laboratory. The plant
material was extracted with a dilute hydrochloric acid
solution and filtered. The filtrate was then basified and
the analyte extracted into chloroform for identification
by gas chromatography-mass spectrometry (GC-MS)
Fig. 2: GC-MS chromatogram of plant material extracts
[1, 2]. GC-MS chromatogram showed a major peak at
R.T =5.77 min (Fig 2) and our mass spectra library
search result matched cathinone (Fig. 3). The plant
was identified as khat (Catha Edulis) which contained
the controlled substance cathinone [1-3], a compound
that is not found in other members of the genus [4].
This is the laboratory’s first encounter of khat.
Fig. 3: Mass Spectrum of cathinone showing m/z 44 as the
base peak
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
Several drums of a light brownish substance labeled
“sodium D-pantophenate” in a very large
methamphetamine
clandestine
laboratory was
identified by GC-MS and FTIR as αacetylphenylacetonitrile (Fig. 4 and Fig. 5). The IR
spectrum of the unknown (Fig. 6) matched the αacetylphenylacetonitrile reference IR spectrum from
the HR Toronto Forensic Library.
Fig. 4: TIC of unknown (Top) and MS of the peak at 6.67
min (Bottom).
Fig. 5: MS of peak at 6.76 min (Top) and MS of αacetylphenylacetonitrile from NIST library (Bottom)
Case 5 (2006)
Fig. 6: IR spectrum of the unknown (α-acetylphenylacetonitrile)
Pure α-acetylphenylacetonitrile has a melting point of
86-89 °C [5]. This chemical, also known as αacetylbenzylcyanide, can be easily converted to
phenyl-2-propanone, a common precursor for the
synthesis of methamphetamine [6]. As expected from
the utilization of this precursor the methamphetamine
hydrochloride produced at this clandestine laboratory
was determined by our in-house chiral HPLC
procedure to be a racemic mixture of the dextro and
levo enantiomers (Fig. 7).
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
Fig. 7: HPLC trace showing the racemic methamphetamine HCl found at the clandestine laboratory.
Case 6 (2010)
Some dried stringy plant material (Fig. 8) suspected
by the law enforcement to contain a controlled
substance was submitted to the laboratory for analysis.
On examination the plant material was found to be
“ma huang” or ephedra. A GC-MS screen confirmed
the presence of ephedrine/pseudoephedrine and
methylephedrine [8].
Fig. 8: Dried “ma huang” (Ephedra)
b) The Black Pearl (2010)
The practice of body packing is defined as the
trafficking of illicit drugs within the gastrointestinal
tract or vagina. In view of the difficulty of its detection
and the potentially large financial gain it is a popular
way of smuggling drugs. Body packers are also called
“swallowers”, “internal carriers” and “mules”. The
body packer usually carries about 1 kg of the drug,
divided into 50-100 packets of 8-10 g each, although
persons carrying more than 200 packets have been
reported [9]. Packets swallowed are usually around 2
cm in size and spherical in shape while rectally
inserted packets are bigger [10]. We have encountered
quite a number of drug cases involving “swallowers”
in Malaysia since the late 1980s. These cases usually
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
involved heroin, cocaine, and more recently crystalline
methamphetamine hydrochloride. In a recent case, the
police submitted 100 such concealed packets which
were
suspected
to
contain
crystalline
methamphetamine (Fig. 9). These were recovered
from the gastrointestinal tract of an Iranian swallower
detained at the airport and were probably not machine
made as their size and shape were not quite uniform
[11].
Colour tests, GC-MS, FTIR-ATR and gas
chromatography-flame ionization detector (GC-FID)
confirmed the packets (of mean weight ca.12 g) to
contain methamphetamine hydrochloride of about 98%
purity. However, one of the packets was found to
contain a black gummy substance consistent with the
appearance and odour of raw opium. The Marqius and
Froehde reagents tested positive for opiates while the
ferric chloride test indicated the presence of meconic
acid which is present in both prepared and raw opium.
The 2N hydrochloric acid and Thin Layer
Chromatography
(TLC)
tests
(positive
for
porphyroxine) showed the substance to be raw opium.
A GC-MS analysis confirmed the presence of the five
principal opium alkaloids, namely the codeine,
morphine, thebaine, papaverine and noscapine in the
material (Fig. 10).
In addition, the relatively high papaverine content of
this raw opium sample indicated that it was not of the
South East Asian type (Fig. 11) and had probably
originated from another region. From our experience,
raw opium from Iran (which shares a border with
Afghanistan) has a relatively high papaverine level.
This finding thus corroborated the law enforcement’s
information that the concealed drugs originated from
Iran. This is the first case to have found raw opium in
a body packer since it is rarely reported elsewhere due
its considerably lower financial value compared to
drugs like methamphetamine, cocaine and heroin.
Fig. 10: GC-MS chromatogram of the concealed raw opium
showing a pronounced papaverine peak at R.T = 14.003 min
Fig. 11: GC-MS chromatogram of raw opium of South East
Asian origin showing a papaverine peak with low intensity.
Conclusion
We report some most recent cases involving the
analysis of unknown materials submitted to our
laboratories. This reflects the nature of the work in our
laboratory. It also delineates the importance of
experience and common sense in the drugs laboratory
and how the information obtained from various
analyses can help in law enforcement investigations.
They help illustrate that drug analysis can be an
exciting and challenging field.
References
1.
2.
Fig 9: Some of the packets recovered from gastrointestinal
tract (markings made by investigating officer)
3.
M Lee. (1995). The identification of cathinone in
khat (Catha Edulis): A time study. Journal of
Forensic Sciences. 40(1): 116-121.
J Chappell, M Lee. (2010). Cathinone
preservation in khat evidence via drying. Forensic
Science International. 195(1): 108-120.
T. Lehmann, S. Geisshüsler, R. Brenneisen.
(1995). Rapid TLC identification test for khat
(Catha edulis). Forensic Science International.
45(1-3): 47-51.
10
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
4.
5.
6.
7.
8.
9.
L. Ripani, S. Schiavone, L. Garofano. (1996).
GC/MS identification of Catha edulis stimulantactive principles. Forensic Science International.
78(1): 39-46.
http://chemfinder (Assessed: 10 April 2010).
Mike White. (2004). FSS Report on
methamphetamine chemistry, seizure statistics,
analysis, synthetic routes and history of
methamphetamine manufacture in UK and the
USA for the ACMD working group.
Y Makino et al. (2002). Impurity profiling of
ephedrines by HPLC. Journal of Chromatography
A. 947: 151-154.
JF Cui, TH Zhou. (1991). Analysis of alkaloids in
Chinese Ephedra species by gas chromatographic
methods. Phytochemical Analysis. 2: 116-119.
S.J. Traub, RS Hoffman, LS Nelson. (2003). Body
packing – the internal concealment of illicit drugs.
New England Journal of Medicine. 349: 25192526.
10. Hergan K, Kofler K, Osier W. Drug smuggling by
body packing: what radiologists should know
about it. European Radiology. 736-742.
11. N. Bulstrode, F. Banks, S. Shrotria. (2002). The
outcome of drug smuggling by 'body packers' -the
British experience. Annals of the Royal College of
Surgeons of England. 84: 35–38.
Additional information and reprint requests:
Chan Kee Bian
(Email: kbchan@kimia.gov.my)
Narcotics Section
Forensic Division
Chemistry Department of Malaysia
46661 Petaling Jaya, Selangor, Malaysia
11
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
Determination of Formaldehyde Contamination in Wiping Media
Used in Trace Pseudoephedrine Analysis
Ahmad Fahmi Lim Abdullaha, Gordon M Miskellyb
a
Forensic Science Programme, School of Health Sciences, Universiti Sains Malaysia, Kelantan, Malaysia
Forensic Science Programme, Department of Chemistry, The University of Auckland, Private Bag 92019,
Auckland, New Zealand
b
ABSTRACT: Pseudoephedrine can be used to measure the level of contamination and to evaluate the
effectiveness of decontamination for surfaces at a clandestine laboratory which has been contaminated during its
operations. Surface wiping using appropriate wiping media have been suggested by cleanup guidelines. Some
wiping vehicle used for low levels of pseudoephedrine showed possible formaldehyde contamination that leads
to the detection of a pseudoephedrine-formaldehyde adduct upon GC-MS analysis. A spectrofluorometric
method based on the Hantzsch reaction was used to quantify extractable formaldehyde from cotton wool,
Whatman 40 and Sartorious filter papers. Formaldehyde was initially present in all the wiping media at levels up
to almost 1.0 g per filter, but a detergent wash greatly reduced the level of formaldehyde contamination.
Formaldehyde levels slightly increased when washed and dried wiping media were exposed to laboratory air for
a period of 18 h to 48 h, whereas a control in a clean sealed bottle showed no increase. Therefore, materials used
as surface wiping media must be properly treated or protected against formaldehyde contamination if
pseudoephedrine is to be recovered and analysed underivatised using GC-MS.
Keywords: formaldehyde, pseudoephedrine, wiping media, contamination, surface
Introduction
Pseudoephedrine [1] is one of the more common
starting materials for clandestine methamphetamine
synthesis, especially via the red phosphorus method
[2, 3]. Although methamphetamine is the principal
indicator for contamination assessment at a
clandestine laboratory, pseudoephedrine can also
be used for this role, especially when has been used
as the sole precursor in a methamphetamine
synthesis or in a situation when the synthesis
process has not completely converted the precursor
to methamphetamine. Since extraction of
pseudoephedrine from pharmaceutical preparations
may be performed in a separate precursor chemical
extraction laboratory from where the actual
methamphetamine synthesis occurs, possibly as a
means to avoid being detected by law enforcement
personnel, the presence of only pseudoephedrine on
surfaces of a suspected clandestine laboratory may
indicate that it was an extraction laboratory.
Upon remediation of a former clandestine
laboratory, pseudoephedrine can be used in
addition to methamphetmine as a surrogate analyte
to measure the effectiveness of decontamination,
and therefore to aid in the determination of whether
the property is considered adequately clean and
safe for subsequent occupancy. Overseas
clandestine
laboratory
cleanup
guidelines
recommend wipe samples to be taken from various
surfaces of the structure before and after the
cleanup when necessary [e.g. 4, 5, 6]. This implies
that surface wiping is a vital step in the evaluation
of the level of contaminants actually present [7]. A
few criteria must be met when selecting a wiping
vehicle – it must hold solvent, its matrix should not
interfere with GC-MS identification, and it should
not fall apart during the extraction process.
Whatman 40 filter paper, Sartorius 1388 filter
paper, cotton wool, and WEBCOL® skin cleansing
alcohol swabs were potential candidates as wiping
vehicles. However, when low levels of
pseudoephedrine were recovered from such media
and analysed underivatised by GC-MS in our
laboratory, the results indicated there might be
contamination from the wiping media since the
chromatograms showed a peak with similar
retention time and mass spectrum to that reported
earlier for the pseudoephedrine-formaldehye
derivative [8-10]. Several authors have reported
that pseudoephedrine or ephedrine can react with
aldehydes or ketones to form oxazolidines [8, 9,
11]. The formaldehyde-based oxazolidines have
almost identical retention times to the parent drugs,
but their mass spectra show a base ion at m⁄z 71
rather than the m⁄z 58 characteristic of the parent
drugs when analysed underivatised by GC-MS [7,
10]. Formaldehyde is a ubiquitous contaminant,
and so wiping media such as filter papers may
contain or accumulate formaldehyde.
12
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
We conducted a study to qualify and quantify
formaldehyde present in the wiping media based on
the Hantzsch reaction of formaldehyde and
acetylacetone (2, 4-pentanedione) in the presence
of ammonia to give a di-acetyldihydropyridine (Fig.
1). The di-acetyldihydropyridine fluoresces so that
trace formaldehyde can be determined using a
spectrofluorometer [12].
Fig. 1: Hantzsch reaction
Materials and Method
All chemicals were of analytical grade and were
used as received. Water for solution preparation
and rinsing of wiping media was from a Milli-Q
purification system. Wiping materials selected
were Whatman 40 filter paper, Sartorius 1388 filter
paper, and cotton wool. WEBCOL® skin cleansing
alcohol swabs were excluded from further
investigation as it is wetted with iso-propanol.
The procedure used for the fluorometric analysis of
formaldehyde was based on that reported by
Belman [12].
(i) A working solution for the analysis was
prepared by mixing equal volumes of 2.0
mol/L ammonium acetate solution (adjusted
to pH 6) and 0.02 mol/L acetylacetone
solution (adjusted to pH 6). Standards in the
range 0 – 2.0 µg of formaldehyde were
prepared by adding 5.0 mL of the working
solution, the appropriate volume of 1.0
mg/mL formaldehyde solution, and making
the volume to 10.0 mL with water. Each
solution was then mixed, capped, and heated
at 37-38°C for 1 h. All the tubes were then
cooled to room temperature prior to analysis.
A Hitachi F-2000 spectrofluorometer was
used for the analysis with ex = 410 nm, em =
515 nm, and 10 nm bandwidth.
(ii) Sample preparation
Triplicates of cotton wool, Whatman 40 filter
paper, and Sartorius 1388 were prepared (see
sections (a), (b) and (c)): 5.0 mL of the above
working solution were then transferred into
each of nine clean dry test tubes. Cotton wool
(about 0.5 g) was put into each of three tubes,
two pieces of Whatman 40 filter paper (5.5
cm in diameter) were put into each of three
tubes, and two pieces of Sartorius 1388 (5.5
cm in diameter) were put into each of three
tubes. All the tubes were made to 10.0 mL
with water, capped and heated at 37-38°C for
1 h, they were then cooled to room
temperature
before
measuring
the
fluorescence response.
a) Measurement of formaldehyde in untreated
cotton wool and filter papers
Measurement of formaldehyde in untreated
cotton wool (ca 0.5 g), Whatman 40 filter
paper (5.5 cm in diameter) and Sartorius 1388
filter paper (5.5 cm in diameter) were
performed as described above.
b) Washed cotton wool and filter paper
To remove any formaldehyde in the cotton
wool, Whatman 40 and Sartorius 1388 filter
papers, they were soaked in 5% Decon 90 for
30 min, sonicated for 10 min, then rinsed
thoroughly with Milli-Q water, and dried at
50ºC in an oven. They were then reanalysed
for formaldehyde as described in (ii).
c) Effect of exposure of Sartorius 1388 filter
paper to laboratory air
Sartorious 1388 filter papers were cleaned
and dried as described above, then they were
exposed in two separate laboratories with a
control in a sealed glass bottle over a period
of 18 h to 48 h. The filter papers were then
analysed for formaldehyde as described in (ii).
Results and Discussion
The calibration curve was linear over the range 0 to
0.2 µg formaldehyde in 10 mL solution.
Formaldehyde was present in all the three sampling
media in quantities up to almost 1.0 g per filter,
Table 1, with Sartorius 1388 filter paper having the
most formaldehyde. The presence of formaldehyde
in some paper products has been reported
previously [13, 14]. The Decon 90 wash reduced
13
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
the amount of formaldehyde in all samples,
particularly for the Sartorius 1388 filter paper,
Table 1.
The effect of exposure to formaldehyde in the
laboratory atmosphere was also examined.
Sartorious 1388 filter paper was chosen for further
investigation as it showed the least amount of
formaldehyde after the washing process, plus it is
durable to the washing and the base extraction
process used for pseudoephedrine wipe analysis
(with 4% NaOH solution and n-hexane). Exposure
of cleaned Sartorius 1388 filters to a new (< 2 years)
laboratory and an old laboratory led to slight
increases in formaldehyde detected in the exposed
samples from both laboratories, whereas the control
showed no increase, Table 2. The sample labeled
“re-used” in the table refers to Sartorius 1388 filter
papers that had been previously used for
pseudoephedrine extraction, which had then been
cleaned and dried before performing the
formaldehyde determination. Only a trace amount
of formaldehyde was detected in this sample.
Table 1: Amount of formaldehyde in the tested wiping materials
Amount (µg)
Untreated
Washed
0.254
0.093
0.361
0.188
0.858
0.040
Samples
Cotton wool (0.5 g)
Whatman 40 (2 pcs of 55 mm diameter)
Sartorius 1388 (2 pcs of 55 mm diameter)
Table 2: Amount of formaldehyde in the tested wiping materials after exposure to ambient air
Sartorius 1388 Filter Paper
Untreated (control)
Clean and dried (control)
Clean and dried , exposed 18 h in old laboratory
Clean and dried , exposed 18 h in new laboratory
Clean and dried , kept 18 h in capped bottle (control)
Clean and dried , exposed 48 h in old laboratory
Clean and dried , exposed 48 h in new laboratory
Clean and dried , kept 48 h in capped bottle (control)
Re-used, clean and dried
We
have
reported
earlier
that
when
pseudoephedrine-containing samples that has been
exposed to formaldehyde were analysed
underivatised using GC-MS, a peak eluted at a
Amount (µg)
0.893
0.106
0.236
0.203
0.089
0.258
0.276
0.107
0.058
retention time which was extremely close to that of
pseudoephedrine but that had m/z 71 as its base
peak [10], Fig. 1, rather than the m/z 58 which is
expected for pseudoephedrine [15], Fig. 2.
Fig. 1: GC-MS mass spectrum of pseudoephedrine-formaldehyde adduct
14
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
Fig. 2: GC-MS mass spectrum of pseudoephedrine
This altered GC-MS behavior of pseudoephedrine
in the presence of a formaldehyde source has
previously been reported to have led to the
misidentification
of
pseudoephedrine
as
phenmetrazine [8]. Furthermore, Lewis et al. [9]
have reported that the presence of formaldehyde in
solvents or specimens during pseudoephedrine
urinalysis leads to oxazolidine formation. Thus,
formaldehyde-contaminated surfaces or wiping
materials can lead to the observation of the
formaldehyde adduct of pseudoephedrine when
wipe samples are analysed [7, 10]. The formation
of the pseudoephedrine-formaldehyde adduct could
occur via the reaction pathway in Fig. 3 as has been
proposed earlier [9, 10].
H
OH
H
H
OH
N
CH 3
H
+ 
CH3
H
-O

N
O
CH3
H
Pseudoephedrine
CH3
Formaldehyde
-H
H
1
2
O
H
OH
5
N
3
4
-H
CH 3
H2 O
N
-H2O
CH 3
CH3
CH3
(+)-3,4-Dimethyl-5-phenyl-1,3-oxazolidine
Fig. 3: Reaction of pseudoephedrine with formaldehyde
15
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
Conclusion
Since formaldehyde is potentially present in built
environments,
sample
collecting
vehicles,
glassware and solvent used in pseudoephedrine
extraction must be protected against formaldehyde
contamination. Our results show that Sartorius
1388 filter paper was suitable as the wiping vehicle
for pseudoephedrine surface recovery experiments,
although similar filter papers should also be
suitable following appropriate washing to remove
formaldehyde. When used for collecting low level
pseudoephedrine intended for GC-MS analysis,
wipe materials should be pre-cleaned with Decon90, dried and wrapped with aluminium foil prior to
use.
7.
8.
9.
Acknowledgements
Mr Glenn Boyes is thanked for the technical advice
in fluorometric analysis, and scientists from ESR
Ltd. for useful advice and material support.
10.
References
11.
1.
2.
3.
4.
5.
6.
Salocks, C. and K.B. Kaley. (2003). Technical
Support Document: Toxicology Clandestine
Drug Labs/ Methamphetamine. Ephedrine and
Pseudoephedrine.
1(13):
www.oehha.ca.gov/public_info/pdf/TSD%20E
phedrine%20Meth%20Labs%2010'8'03.pdf.
Skinner, H.F. (1990). Methamphetamine
Synthesis via Hydriodic Acid/Red Phosphorus
Reduction of Ephedrine. Forensic Science
International. 48: 123-134.
Chan, K., et al. (2009). Types of Clandestine
Methamphetamine Laboratories Seized in
Malaysia. Department of Chemistry Malaysia.
Guidance and Standards for Cleanup of Illegal
Drug-Manufacturing Sites (FINAL). (2004).
Alaska
Department
of
Environmental
Conservation:
http://dec.alaska.gov/spar/perp/docs/druglab_g
uidance.pdf.
Clandestine Drug Lab General Cleanup
Guidance (July 1, 2006 Version). (2006)
Minnesota Department of Health, Minnesota
Pollution
Control
Agency:
www.health.state.mn.us/divs/eh/meth/lab/guid
ance0606.pdf.
Clandestine Drug Lab Program, Guidelines for
Environmental Sampling at Illegal Drug
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Manufacturing Sites. 2005, Washington State
Department
of
Health:
http://www.doh.wa.gov/ehp/cdl/guideenvirsamp.pdf.
Abdullah, A.F.L. and G.M. Miskelly. (2010).
Recoveries of Trace Pseudoephedrine and
Methamphetamine
Residues
from
Impermeable
Household
Surfaces:
Implications for Sampling Methods Used
During
Remediation
of
Clandestine
Methamphetamine Laboratories. Talanta. 81:
455-461.
Lambert, E.E.W. (2004). Phenmetrazine or
Ephedrine? Fooled by Library Search. Journal
of Chromatography A. 1045: 259-262.
Lewis, R. (2000). Formation of an Interfering
Substances,
3,4-Dimethyl-5-Phenyl-1,3Oxazolidine, During a Pseudoephedrine
Urinalysis. Journal of Forensic Sciences. 45(4):
898-901.
Miskelly, G.M. and A.F.L. Abdullah. (2009).
Formation of Trifluoroacetylated Ephedrine
During the Analysis of a Pseudoephedrine formaldehyde Adduct by TFAA Derivatization
Followed by GC-MS. Journal of Forensic
Sciences. 54(2): 365-367.
Neelakantan, L. (1971). Asymmetric synthesis
II. Synthesis and Absolute Configuration of
Oxazolidines Derived from (-)Ephedrine and
Aromatic Aldehydes. Journal of Organic
Chemistry. 36: 2256-60.
Belman, S. (1963). The Fluorimetric
Determination of Formaldehyde. Analytica
Chimica Acta. 29: 120-126.
Fisher, A.A., N.B. Kanof, and A.M. Biondi.
(1962). Free Formaldehyde in Textiles and
Paper -Clinical Significance. Archieves of
Dermatology. 86: 753-756.
Möller, B. and A. Hensten-Pettersen,
Biological Evaluation of Absorbent Paper
Points. International Endodontic Journal. 18
(3): 183-186.
Smith, R.M. (2005). Understanding Mass
Spectra: A Basic Approach. 2 ed. Hoboken,
N.J: Wiley Interscience.
Additional information and reprint requests:
Ahmad Fahmi Lim bin Abdullah
(E-mail: fahmilim@kb.usm.my)
Forensic Science Programme
School of Health Sciences
Universiti Sains Malaysia
16150 Kubang Kerian, Kelantan, Malaysia
16
Malaysian Journal of Forensic Sciences, 2010, Vol 1
Forensic Light Sources for Detection of Biological Evidences in
Crime Scene Investigation: A Review
Wee-Chuen Leea, Bee-Ee Khooa
a
School of Electrical and Electronic Engineering, Universiti Sains Malaysia, Penang, Malaysia
ABSTRACT: Identification of biological evidences, such as blood, semen, saliva and urine, are important for
crime scene investigation. Forensic light sources have been used for detection of biological evidences, where
this method is a simple, presumptive, non-destructive test and applicable for detecting most types of biological
evidences. Biological evidences can be detected by forensic light source due to their natural characteristic, such
as light absorption (blood) or fluorescence effect (semen, saliva and urine). Biological evidences on different
materials would have different effect in detection, where materials with high absorbent or exhibit strong
fluorescence would affect the detection of biological evidences. This paper reviews on the methods and
limitation of detecting biological evidences by forensic light source and provide the recommendations for
improving the detection techniques using forensic light source.
Keywords: Forensic light source, biological evidence, blood, semen, urine, saliva
Introduction
Biological evidences such as blood, semen, saliva
and urine are among the most important evidences
in crime scene investigation [1]. Valuable
information can be obtained from the biological
evidences found, such as DNA evidence for the
identification of the victims and suspects and the
bloodstain pattern for the determination of the
sequence of events [1]. Several methods have been
developed for the identification of these biological
evidences, which can be divided into presumptive
tests and confirmatory test. Presumptive tests are
just screening tests, whereas confirmatory tests will
conclusively identify the species of the particular
evidence [1]. However, most of the tests are
destructive test, where the DNA evidence would be
destroyed, and some tests can only be carried out in
a laboratory [1]. One of the simplest presumptive
tests that can be used to determine most of the
biological evidence is forensic light source (FLS)
[1].
FLS is a term used commonly to refer to an
illumination system adapted in forensic application,
such as laser and high-intensity filtered lamps [2].
A non-laser FLS is sometimes referred as an
alternative light source (ALS) [2]. FLS can either
make the evidence fluorescence or enhance the
contrast of the evidence against the background [3].
Fluorescence happens when the FLS emitted to the
biological evidences, such as semen, saliva and
urine, where these fluids absorbed light at
particular wavelength and then re-emits the
absorbed energy as light at a longer wavelength [2].
Besides, FLS can be used to enhance contrast of
bloodstain on dark surfaces, where the stain is not
visible to naked eye, such as bloodstain [3].
The maximum detectable dilution of biological
stains by FLS was relatively lower compared to
chemical based method, such as luminol for
bloodstains, where maximum detectable dilution of
bloodstains was 1/1000 using Polilight® [4],
whereas luminol sensitivity was up to 1/5000000
[5]. However, since no chemical was required and
it was easy to use, FLS was commonly used in
crime scene investigation as a scanning tool.
Besides, most of the FLS was used to enhance the
biological stains towards its background for
photography purposes. Moreover, FLS was suitable
in detecting various types of biological evidence
[1].
This paper reviews the methods of detection using
FLS, brief overview of different types of available
FLS and also the effects of biological evidences on
different surfaces towards the detections. This
paper is organized as follows:
 Introduction of the responses of biological
evidences, such as blood, semen, saliva and
urine towards different light wavelength of
FLS.
 Discussion of the detection of biological
evidences using different types of FLS. The
comparisons between each FLS that have been
reported in literature in discussed in this
section.
 Summary of the effects of biological evidences
on different surfaces towards their detections
using FLS. This section reported the maximum
17
Malaysian Journal of Forensic Sciences, 2010, Vol 1
detectable dilution of biological evidence on
different surfaces using FLS that were found in
literature.
Responses of Biological Evidences towards FLS
absorb certain wavelength of light, excitation
spectrum, and re-emit a longer wavelength of light,
emission spectrum [2,4]. Stoilovic [6] reported that
the emission spectrum of semen stain was covering
the region of 400nm-700nm with the excitation
spectrum measured in 300-480nm by Polilight®, as
shown in Fig. 2.
(a) Blood
Untreated dry blood does not show a significant
fluorescence effect but it has a high absorption in a
very broad region of light wavelength from 300900nm, which cover the entire light wavelength,
including UV, visible (VIS) and IR light [6]. Hence,
bloodstain will occur as a dark spot when it was
exposed to any type of light. Most of the FLS were
able to enhance the contrast of bloodstain towards
its background, especially on dark background [113]. Bloodstain would appear to be brighter against
a dark background [2].
Along the high absorption region, the strongest
absorption band occurred in a narrow band of
395nm to 435nm, with the maximum absorption at
415nm due to the presence of haemoglobin, as
shown in Fig. 1 [6]. Background correction method
was proposed to further enhance the dry untreated
blood towards the background using this narrow
wavelength of light [11,12]. In their experiment,
the light source used was Rofin PL-10 Polilight,
which is a high intensity xenon lamp with
selectable narrow bandpass filters. These bandpass
filters can be adjusted to emit the light with peak at
435nm, 415nm and 395nm. Due to the difference
in absorption between 395nm, 415nm and 435nm
light, this method can enhance the bloodstain
images that are unclear even at 415nm, where the
background exhibits almost the same absorption of
light [11,12].
Fig. 1: Absorption Spectrum of Dry Blood [6]
(b) Semen
Untreated dry semen is a very strong
photoluminescence substance [6], where it would
Fig. 2: Photoluminescence spectra of dry untreated
semen [6]
By supplying the specific excitation light with
appropriate goggles or filters, semen stain can be
clearly observed due to the photoluminescence
effect of semen. Goggles were used to filter out
strong excitation wavelength and only allowed the
emission wavelength to pass through [6]. Table 1
summarized the procedures for detection of
untreated semen stain using different types of
excitation light with the appropriate goggles or
filter, which was reported by Stoilovic [6]. The test
was run from the combination of UV light without
goggles to the combination of green-yellow light
with violet filter. Suitable light with goggles
combination was chosen when the background was
not photoluminescence during observation [6].
In a more recent paper reported by Nelson and
Santucci [14], a test to determine the best
combination exciting wavelength and goggles for
viewing semen stains was done using Omniprint
1000, an adjustable wavelength light source with
narrow band increments (30-40nm) between
320nm to 510nm and different coloured goggles,
which were yellow, orange and red goggles. The
best fluorescence effect of semen stains was found
at the wavelength of 420nm and 450nm, observed
with human eyes through orange goggles. It was
also reported that semen stains would appear as a
yellow-greenish stains when exposed to a
continuous green beam at 532nm wavelength.
These continuous green beam was generated from a
laser’s type FLS, Spectra-Physics® RevealTM, and
the semen stains observed through a orange laser
safety goggles that block 532nm wavelength [9].
18
Malaysian Journal of Forensic Sciences, 2010, Vol 1
Table 1: Excitation light with appropriate goggles for untreated semen stains detection recommended by Stoilovic [6]
Excitation light
UV
Violet
Blue
Green
Green-yellow
Green-yellow
Goggles/Filters
No goggles needed, but recommended to wear UV safety goggles
Yellow goggles
Yellow goggles
Orange goggles
Red goggles
Violet filters (425nm)
(c) Saliva
Dried saliva stain is virtually colourless and
difficult to detect by naked eye [15]. From
literature, saliva stain exhibits fluorescence effect
but in a lower intensity compared to semen [15].
Saliva stain was detectable by naked eye when
exposed to UV light [9], where it would appear
bluish-white, but this would not differentiate it
from other stains [1,16]. In addition, UV-UV
photoluminescence, where excitation wavelength at
short UV (266nm) and emission wavelength at long
UV, was reported to be able to detect saliva stain
[10]. Besides, saliva stain was also detectable under
excitation wavelength of 450nm with orange
goggles [4] or 555nm interference filters, which is
a filter that use interference effect to transmit
555nm wavelength of light and reflect other
wavelengths [15]. Camilleri et al. [15] reported that
the optimum contrast of saliva stains on white
Colour of the observed stain
Blue
Yellow
Yellow
Orange
Red
Black
cotton background was achieved using the 470nm
excitation wavelength with the 555nm interference
filters, while saliva stain was also detectable with
human eyes using other excitation wavelength with
different colour goggles or filters, such as 415nm
with yellow goggle or 555nm interference filters,
470nm with 530nm interference filters, 490nm with
555nm interference filters and 505nm with 555nm
interference filters. Furthermore, saliva stain
appeared to be white thin edged stain when
exposed to 450nm excitation wavelength and
viewed with orange goggles [4]. Besides, yelloworange stain was observed when saliva stain
exposed to 532nm excitation wavelength and
viewed with goggles that designed to block 532nm
light [9]. Table 2 summarized the excitation
wavelength with the suitable goggles or
interference filters for detection of saliva by human
eyes.
Table 2: Excitation wavelength with suitable goggles or filters for detection of saliva
Excitation light
Long UV [1,9,16]
415nm [4,15]
450nm [4,15]
470nm [15]
490nm [15]
505nm [15]
532nm [9]
Goggles/Filters
No goggles needed, but recommended to wear UV safety goggles
Yellow goggles/555nm interference filters
Orange goggles/555nm interference filters
530nm/555nm interference filters
555nm interference filters
555nm interference filters
Goggles that block 532nm light
Colour of the observed stain
White-bluish
Not stated in literature
White (Orange goggles)
Not stated in literature
Not stated in literature
Not stated in literature
Yellow-orange
Notes: interference filters allow only the desirable wavelength pass through.
(d) Urine
Urine stains are hard to be seen because the nature
of urine, where these stains will become diluted on
fabric surfaces [1]. In fact, urine stains exhibits
fluorescence effect when exposed to UV light, but
the colour of the stain may vary in the presence of
abnormal substances, such as glycosuria [16].
Vandenberg and Oorschot [4] reported that urine
was detectable by human eyes under 415nm
excitation wavelength with yellow goggles, 450nm
excitation wavelength with orange goggles and
505nm excitation wavelength with red goggles.
Besides, Seidl et al. [9] tested urine stains with
excitation wavelength at 532nm using SpectraPhysics® RevealTM. From the results of their test,
when urine stains were viewed under goggles that
block 532nm light, the stain appeared as a yelloworange stain but more intense compared to saliva,
which was also exhibits the same colour under this
wavelength/filters
combination.
Table
3
summarized the excitation wavelength with the
suitable goggles or filters for detection of urine.
19
Malaysian Journal of Forensic Sciences, 2010, Vol 1
Table 3: Excitation wavelength with suitable goggles or filters for detection of urine
Excitation light
UV [16]
415nm [4]
450nm [4]
505nm [4]
532nm [9]
Goggles/Filters
No goggles needed, but recommended to wear UV safety
goggles
Yellow goggles
Orange goggles
Red goggles
Goggles that block 532nm light
Detection of Biological
Different Type of FLS
Evidences
using
There were several FLS reported for the success in
aiding the detection biological evidences by human
eyes. In order to increase the sensitivity of
detection, an FLS must produce a high intensity of
light, as Wawryk and Odell [13] reported that most
FLS with lower intensity was not suitable to be
used to detect urine due to its weak emission light.
Besides, different substances have different
excitation wavelengths that would give the best
detection. Due to these two circumstances, among
all the available FLS, the most tested FLS in
literature was Polilight®, where this FLS gives
variety of wavelength with high intensity of light.
Vandenberg and Oorschot [4] reported the test of
the latest Polilight®’s model, PL500, on blood,
semen, urine and saliva and it shows positive
detection for all type of tested stains when
observed through suitable goggles.
In fact, lasers, such as TracER and SpectraPhysics® RevealTM, have higher intensity and a
narrower bandwidth compared to most of the FLS,
which make it to be more effective in detection
compared to other light source [3]. In an earlier
paper, Auvdel [17] reported that a laser, SpectraPhysic Model 171-19, was more effective in
detecting semen, saliva and sweat stains compared
to Mineralight, short UV light source with 245nm.
However, in another paper reported by Auvdel [18],
high-intensity quartz arc tube, Luma-Print, has a
better detection rate of semen, saliva and sweat
compared to laser, Spectra-Physic Model 171-19.
Furthermore, Luma-Print has a better portability
over high-powered laser. In addition, lasers are
more costly and heavier compared to other FLS [3].
The comparisons of the detection blood, semen,
saliva and urine between Lumatec Superlite 400, a
tunable wavelengths FLS with several different
goggles, and Spectra-Physics® RevealTM, with
goggles that block 532nm light emitted from the
laser, were reported by Seidl et al. [9]. Both light
sources showed comparable results in detection but
fluorescence of urine was stronger with laser
system. However, blood stain cannot be viewed
using the laser system of 532nm wavelength with a
safety goggles that block 532nm light [9]. The
Colour of the observed stain
Depends on abnormal substance presence
Not stated in literature
White
Not stated in literature
Yellow-orange
contrast between blood stains with the background
cannot be enhanced as blood stains absorbed
completely the light from laser and the light
reflected from the background was blocked by the
safety goggles.
Due to the strong absorption wavelength of blood
stain in the entire light wavelength, there were
several types of suitable FLS for bloodstains’
detection reported in literature, such as high
intensity LED [13], UV [8,10], Lumatec Superlite
400 [9], PolirayTM [13] and Polilight® (1-2; 4-6;
11-12). Besides, IR light was also proven to be
successful in detecting bloodstain on black fabrics
[7]. Moreover, it was reported that bloodstain that
was covered by paint can also be revealed using
Polilight® [4].
Santucci et al. [19] reported that Wood’s Lamp, an
ultraviolet light source that emits wavelengths of
approximately 320-400nm, was unable to be used
to distinguish between semen and other substances
that were commonly found on perineum of children
or adolescents. Moreover, all the 29 semen samples
used for the study did not show any fluorescence
when exposed to Wood’s Lamp. Bluemaxx BM500,
a FLS with a broad-band wavelength of 390-500nm
was tested and reported to have a better
performance compared to Wood’s Lamp [14].
Using Bluemaxx BM500, semen stain on a white
100% cotton surface can be detected all of the time
and after a brief training session, 15 out of 18 of
the physicians, about 83.3%, were able to
differentiate semen stain from other common
product. With this FLS, the semen sample would
still exhibit fluorescence with the same intensity
after a few months from the initial placement on
the cloth [14]. PolirayTM with 550nm camera filter
was also tested to be successful in detecting semen
stains by Lincoln et al. [20].
The detection of blood, semen, saliva and urine by
different type of LED, ranging from 370 to 480nm,
and PolirayTM on skin was tested by Wawryk and
Odell [13]. Semen and blood stains were
successfully detected by all of the tested FLS.
However, a very close distance between those FLS
to the surface was needed, which was less than 3cm.
In addition, those stains can only be observed with
filter goggles in a distance around 20cm from the
20
Malaysian Journal of Forensic Sciences, 2010, Vol 1
stains. Besides, saliva was not fluorescence under
any type of the tested light source. This shows
those tested FLS were having lower light intensity
output compared to Polilight®, which caused saliva
undetectable, as saliva shows weak fluorescence
under Polilight®. Besides, urine was not detected
by any lower intensity LED or everLEDTM
MagliteTM replacement bulb, but some fluorescence
observed when urine exposed to LuxeonTM Star V
LEDs and Poliray® [13].
There was another test by Carter-Snell and Soltys
[21] to compare different wavelength effect on
biological stain, such as semen, urine and saliva,
using Mineralight, Evident Products CE, long UV
light with 365nm, Bluemaxx BM500 and
Bluemaxx Mini, blue light with 450nm. Semen
stains were detectable by all the tested FLS good
results, while urine was only detectable using
Mineralight and Bluemaxx BM500, with weak
sensitivity. Saliva stains were also fluorescence
when exposed to each of the tested FLS except for
Bluemaxx Mini, due to its lower intensity of light.
However, the fluorescence of the same semen stain
was observed to be different in colour by different
examiner with the same FLS and goggles. Blue,
blue-white and light green fluorescence colours of
semen stain were observed when exposed to
Evident Product CE, whereas white, yellow-white
and green fluorescence colours were observed
when exposed to Bluemaxx BM500 and Bluemaxx
Mini with orange goggles.
Table 4 summarizes the wavelength output and the
detectable stains reported for all of the FLS found
in recent literature, such as TracER [3], SpectraPhysics® RevealTM [9], PolirayTM [13,20],
Polilight® PL500 [4], Lumatec Superlite 400 [9],
Wood’s lamp [19], Bluemaxx BM500 [14,21],
Bluemaxx Mini [21], Evident Product CE [21],
Mineralight® [21] and high intensity LED [13].
Table 4: Comparison of the tested FLS in recent literature in term of wavelength and detectable stains reported
FLS
TracER (Laser) [3]
Spectra-Physics® RevealTM (Laser) [9]
PolirayTM [13,20]
Wavelength (nm)
532 (Green laser beam)
532 (Green laser beam)
415-610 (mounting interference filters)
Detectable Stains Reported
Semen, Saliva, Urine
Semen, Saliva, Urine
Blood, Semen, Urine
Polilight® PL500 [4]
Adjustable from UV, 415-650nm and
white light
Adjustable from 320-700
320-400 (Long UV)
450 (Blue)
450 (Blue)
365 (Long UV)
254 (Short UV)
Variety of wavelength depends on the
LED used
Blood, Semen, Saliva, Urine
Lumatec Superlite 400 [9]
Wood’s lamp [19]
Bluemaxx BM500 [14,21]
Bluemaxx Mini [21]
Evident Product CE [21]
Mineralight® [21]
High Intensity LED [13]
Effect of Biological Evidences on Different
Surfaces towards their Detection using FLS
The sensitivity of biological stains detection
through human eyes using FLS varies on different
type of surfaces the stains occurred. This was due
to different reactions of different type of surfaces’
materials towards FLS. Some materials were dark
in colour, highly absorbent of liquids or exhibits
strong fluorescence effect when exposed to FLS
[22]. High absorbent material would absorb the
biological evidences into its fabric before it dry and
fluorescence of the background would mask the
fluorescence of the biological stains. These factors
will reduce the contrast enhancement of biological
stain towards background [4]. Table 5 shows the
surfaces that had been tested in literature for the
detection of biological evidences using FLS.
Blood, Semen, Saliva, Urine
Semen (doubtful)
Semen, Saliva, Urine
Semen
Semen, Saliva, Urine
Semen, Saliva, Urine
Blood, Semen (Urine was detectable
by LuxeonTM Star V LED)
(a) Blood
The detection of bloodstain was poor on highly
absorbent polar fleece [4]. The maximum
detectable blood dilution reported was 1/1000,
where the stain was on white cotton and FLS used
was Polilight® PL500 [4]. However, the same
stains was detectable using natural light, which
means FLS has little benefit for detection of
bloodstains on light-coloured surfaces [4].
Wagner and Miskelly [11,12] reported their
background correction method was able to detect
blood dilution up to 1000-1600, with the aid of
Polilight®. They reported that using Polilight® at
415nm, the maximum detectable dilution on white
cotton was 1/400, but the detectable dilution was
further improved by their proposed method.
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Malaysian Journal of Forensic Sciences, 2010, Vol 1
Besides Polilight®, Lumatec Superlite 400,
adjusted to output the light with wavelength of
415nm, was also tested on different type of
materials by Seidl et al. [9] and its maximum
detectable dilutions were shown in Table 6.
Meanwhile, Wawryk and Odell [13] reported that
the bloodstain on skin was detectable using high
intensity LED or PolirayTM, but not visible on the
second day of the experiment. From literature, IR
light was used to detect bloodstain on black colour
surfaces, where the stains were barely visible [7].
The maximum dilutions of bloodstains on different
materials detected by the stated FLS were
summarized in Table 6.
Table 5: Surfaces tested for detection of biological evidences with FLS in literature
Item
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
Surface
Fabric: Cotton [9]
Fabric: Cotton (4; 11-12; 21)
Fabric: Cotton [4]
Fabric: Cotton [4]
Fabric: Cotton [4]
Fabric: Cotton [4,7]
Fabric: Cotton [4]
Fabric: Wool [4]
Fabric: Nylon [4]
Fabric: Nylon [4]
Fabric: Polyester [4]
Fabric: Velour [4]
Fabric: Satin [4]
Fabric: Crepe [4]
Fabric: Polyester + spandex [4]
Fabric: Cotton + elastane [4]
Fabric: Nylon + elastane [4]
Fabric: Polysester + cotton [4]
Fabric: Polar fleece [4]
Synthetic carpet [4]
Pine wood [4]
Dried leaves [4]
Glass [4]
Brick [4]
Metal [4,9]
Plasterboard [4]
Condom [4]
Tile [9]
Glass [9]
PVC [9]
Formica [9]
Carpet [9]
Stone [9]
Wood [9]
Fabric: 35% rayon & 65% polyster [7]
Fabric: 35% cotton & 65% polyster [7]
Fabric: 35% polyster & 65% cotton [7]
Fabric: 100% velvet [7]
Fabric: 50% acrylic & 50% wool [7]
Fabric: 5% lycra & 95% cotton [7]
Fabric: 5% spandex & 95% polyester [7]
Fabric: 30% polyster &70 % rayon [7]
Fabric: 30% acrylic &70% wool [7]
Human skin [13,21]
Colour of surfaces
–
white
red
pink with white polka dot
checked weave
black
yellow, green, pink, blue, brown, purple
white, yellow, green, red, blue
white, blue
pink
white
blue
pink
black
white, black
blue
white
black + white
green
white, blue
–
–
–
–
–
–
–
–
–
–
–
–
–
black
black
black
black
black
black
black
black
black
–
–
Remark: “ –“ not found in literature
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Malaysian Journal of Forensic Sciences, 2010, Vol 1
Table 6: Detectable bloodstains on different surfaces reported using FLS
FLS
Polilight®
Wavelength (nm) / Detection methods
Background Correction [11,12]
415nm [4]
Lumatec Superlite 400
415nm [9]
High Intensity LED
PolirayTM
IR
370-480nm [13]
450nm [13]
>930nm [7]
Code:
Surfaces
2
2
3-18,20-26
19
1
28
29
30
31
32
25
33
34
44
44
6
35
36
37
38
39
40
41
42
43
Maximum visible dilution reported
1/1600
1/1000
–
1
1/10
1/10
1/10
1/100
1/10
1/1
1/1
1/1
1/100
nv2
nv2
1/4
1
1/4
1/4
1/4
1/4
1
1/4
1/8
1/4
Surfaces numbers referred to the surfaces numbered in Table 5
Bold dilutions mean strong or clear detection.
Non-bold dilutions mean weak detection.
“–“ indicates not stated in literature
nv2 means neat stains visible on first day but not visible on second day
(b) Semen
Vandenberg and Oorschot [4] reported that the
most useful wavelength and goggles used for
detection of semen stain was 450nm with orange
goggles. Semen stains can be detected using FLS
on most of the surfaces, where the stains was not
detectable by naked eye under natural light, due to
the strong fluorescence effect of semen stains [4].
Absorbency of fabric was not affecting much the
detection of semen stains, where semen stains on
highly absorbent surfaces, such as blue velour and
dark green polar fleece, were easily detected by
Polilight® [4]. The reported maximum detectable
dilution on white cotton using Polilight® was
1/100 and not detectable under natural light [4].
However, the colour and pattern of the surfaces
was found to affect the appearance of semen stains.
The reported surfaces were pink nylon, red cotton,
pink cotton white polka dot and checked fabrics,
where semen stains on those same materials with
different colour show good results in detection [4].
Fabrics such as white cotton, pink satin and pink
fleecy material, which shows strong fluorescence
under certain wavelength, reduced the contrast
between semen stains and the background [22].
White cotton’s excitation wavelength was about
340-410nm with 440-470nm emission wavelength,
while pink satin’s excitation wavelength was
around 490-530nm with 570-620nm emission
wavelength. Hence, the excitation wavelength with
450nm is better than UV light for detection of
semen stain on white cotton [22]. Moreover, as
reported earlier, Santucci et al. [19] tested that
Wood’s lamp, a long UV light source with the
wavelength about 320-400nm, could not be used to
detect semen stains on white or black cotton.
Another FLS, Bluemaxx BM500, a FLS with the
wavelength of 450nm, showed good results in
detecting semen stains on white cotton [14].
The detection of semen on different surfaces
between Lumatec Superlite 400 with SpectraPhysics Reveal laser was comparable, where the
detectable dilution between both FLS was the same
on every tested surface [9]. Wawryk and Odell [13]
reported that semen stains on skin did not show any
fluorescence when exposed to low intensity LED,
where different LEDs’ wavelengths ranging from
375nm-480nm were tested. When the same stains
on skin exposed to PolirayTM and high intensity
LED, such as LuxeonTM LED, fluorescence was
observed when viewed with orange goggles.
However, the fluorescence was very faint when
23
Malaysian Journal of Forensic Sciences, 2010, Vol 1
exposed to the same FLS on the second day.
Besides, four FLS, Mineralight, Evident Products
CE, Bluemaxx BM500 and Bluemaxx Mini, were
tested and successful in detecting semen stains on
human’s arm [21].
The maximum dilutions of semen stains on
different materials detected by the stated FLS were
summarized in Table 7.
Table 7: Detectable semen stains on different surfaces reported using FLS
FLS
Wavelength (nm) / Detection methods
Polilight®
UV light [22]
450nm + Orange goggles [4]
Bluemaxx BM500
450nm + Orange goggles [14,21]
Mineralight
Evident Products CE
Bluemaxx Mini
Lumatec Superlite 400
254nm [21]
365nm [21]
450nm [21]
415nm + Orange goggles [9]
Spectra-Physics
laser
532nm + 532nm block goggles [9]
High Intensity
(LuxeonTM LED)
Poliray®
Code:
Reveal
LED
452.9nm/466.9nm + orange goggles [13]
Surfaces
2
2,11
3-5,10
6-9,12-27
2
44
44
44
44
1
28
29
30
31
32
25
33
34
1
28
29
30
31
32
25
33
34
44
Maximum visible dilution
reported
1
1/100
1
–
–
–
–
–
–
1
1/10
1/10
1/10
1/100
1/10
1/10
1
1
1
1/10
1/10
1/10
1/100
1/10
1/10
1
1
nf2
44
nf2
450nm + orange goggles [13]
Surfaces numbers referred to the surfaces numbered in Table 5
Bold dilutions mean strong or clear detection.
Non-bold dilutions mean weak detection.
“–“ mean not stated in literature
nf2 means neat stains visible on first day but extremely faint on second day
(c) Saliva
As discussed before, saliva stains are hard to be
detected by naked eye due to its colourless [15].
Moreover, its fluorescence intensity is found to be
very weak and very difficult to be detected by FLS
compared to semen stains [15].
Besides, the absorbency of material became a
factor that affects the detection for saliva stains,
where saliva stains were very difficult to be
detected when the stains were mostly absorbed into
the fabric [4]. Vandenberg and Oorschot [4]
reported that saliva stains on blue and white
checked cotton weave were not visible when
observed through 450nm and orange goggles. The
maximum detectable dilution of saliva stains on
white cotton using Polilight® reported was 1/16
[15].
Wawryk and Odell [13] reported that saliva stains
on human skin was not fluorescence under high
intensity LED, 370-480nm, and PolirayTM, 450nm.
However, it was reported that saliva stains on
human skin was 100% sensitive to UV light,
produced by Mineralight, 254nm, and Evident
Products CE, 365nm, while 14% sensitivity when
using Bluemaxx BM500 [21]. Spectra-Physics®
RevealTM laser has slightly better detection
sensitivity compared to Lumatec Sperlite 400 for
saliva stains [9].
24
Malaysian Journal of Forensic Sciences, 2010, Vol 1
The maximum dilutions of saliva stains on different
materials detected by the stated FLS were
summarized in Table 8.
(d) Urine
Literature on maximum detectable dilution of urine
stains on different surfaces using FLS was limited.
Vandenberg and Oorschot [4] reported that urine
stains on white cotton was detectable with
Polilight® but the serial dilution detection reported
in the literature was only for blood, semen and
saliva stains.
However, the maximum detectable dilution of urine
stains on different surfaces, using Spectra-Physics
Reveal laser and Lumatec Superlite 400, were
reported by Seidl et al. [9], where their detection
capabilities were comparable. Short UV light by
Mineralight and 450nm light by Bluemaxx BM500
were able to detect urine stains on skin, with 71%
and 14% sensitivity respectively [21].
There was also a test of the detection of urine stains
on skin using different type of LED and Poliray®
[13]. However, only higher intensity LED,
LuxeonTM LED and Poliray® were reported to be
successfully in detecting urine stains, with the same
intensity with semen stains. These stains were
undetectable by the mentioned FLS on the second
day of experiment [13].
The maximum dilutions of urine stains on different
materials detected by the stated FLS were
summarized in Table 9.
Discussion
FLS was proven to be successful in detecting
biological stains, such as blood, semen, saliva and
urine. The best single wavelength and goggles
combination for detecting most of the stains was
450nm wavelength with orange goggles [4].
Almost all types of biological stains on white
cotton were visible with this combination of
wavelength and goggles.
Most of the reported detection methods using FLS
were observed through naked eyes. However,
different examiners with the same combination of
FLS and goggles could observe different colour for
the same stains. For example, Carter-Snell et al.
[21] reported that fluorescence of semen stains on
skin were observed by different examiners with
different colours when exposed to the same FLS
and viewed with the same goggles. Moreover,
some of the examiner failed to detect the stains [21].
Hence, there might be failure in detecting
biological stains through human examiner with
FLS during crime scene investigation. Besides,
some fabrics would fluorescence when exposed to
the FLS, such as white cotton would fluorescence
at emission wavelength of 440-470nm with 340410nm excitation wavelength and pink satin would
fluorescence at emission wavelength of 570-620nm
with 490-530nm excitation wavelength [22]. These
fabrics’ fluorescence would make the fluorescence
of stains difficult to be detected. Hence, different
fabric would need different wavelengths with
different interference filters or goggles for the stain
fluorescence to be successfully seen.
Table 8: Detectable saliva stains on different surfaces reported using FLS
FLS
Polilight®
Wavelength (nm) / Detection methods
450nm + Orange goggles [4,15]
Bluemaxx BM500
Mineralight
Evident Products CE
Lumatec Superlite 400
450nm + Orange goggles [14,21]
254nm [21]
365nm [21]
415nm + Orange goggles [9]
Spectra-Physics
Reveal laser
Code:
532nm + 532nm block goggles [9]
Surfaces
2
10,12,19
3-9,11,1318,20-27
44
44
44
28
Maximum visible dilution reported
1/16
1
–
29
30
31
32
25
28
29
31
32
25
1/10
1
1/100
1
1
1/100
1/10
1/100
1
1/10
1
–
–
1/100
Surfaces numbers referred to the surfaces numbered in Table 5
Bold dilutions mean strong or clear detection.
Non-bold dilutions mean weak detection.
“–“ mean not stated in literature
25
Malaysian Journal of Forensic Sciences, 2010, Vol 1
Table 9: Detectable urine stains on different surfaces reported using FLS
FLS
Wavelength (nm) / Detection methods
Polilight®
Bluemaxx BM500
Mineralight
Lumatec Superlite 400
450nm + Orange goggles [4]
415nm + Yellow goggles [4]
505nm + Red goggles [4]
450nm + Orange goggles [21]
254nm [21]
415nm + Orange goggles [9]
Surfaces
2
2
2
44
44
1
Maximum visible dilution
reported
–
–
–
1
–
1/100
1/1000
1/10
1/100
1/1000
1/10
1/100
1/10
1
1/10
1/1000
1/10
1/100
1/1000
1
1/100
1
1
nv2
nv2
Spectra-Physics Reveal
laser
532nm + 532nm block goggles [9]
High Intensity
(LuxeonTM LED)
452.9nm/466.9nm + orange goggles [13]
28
29
30
31
32
25
33
34
1
28
29
30
31
32
25
33
34
44
450nm + orange goggles [13]
44
LED
Poliray®
Code: Surfaces numbers referred to the surfaces numbered in Table 5
Bold dilutions mean strong or clear detection.
Non-bold dilutions mean weak detection.
“–“ means not stated in literature
nv2 means neat stains visible on first day but not visible on second day
Future Developments
Background correction method [11,12], a
multispectral
imaging
algorithm,
showed
improvement in detection of bloodstains. Hence,
there might be improvement of detection of
biological stains using multispectral imaging
system compare to current methods. Multispectral
imaging system is an imaging system that process
images captured in different wavelength.
Multispectral imaging system shows an important
role for enhancing the images in many applications
i.e. food industries [23], medical industries,
printing [24] and forensic [11,12].
Besides, an automated imaging system would aid
the crime scene investigator in detecting biological
evidence. Since there would be different rate of
detection of biological evidence by human
observation, a computerized detection system with
the aid of camera and FLS would give a more
precise detection.
Further investigation for multispectral imaging
system and automated detection system for
detecting various types of biological stains can be
done to improve the detection of biological stains
using FLS.
Acknowledgements
The authors would like to thank University Sains
Malaysia for the financial support through
Postgraduate Fellowship Scheme and Ministry of
Higher Education, Malaysia through FRGS grant
(203/PELECT/6071188).
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Forensic Science International 66: 89-94.
J.H. Wagner, G.M. Miskelly. (2003).
Background
correction
in
forensic
photography I. Photography of blood under
conditions of non-uniform illumination or
variable substrate colour - Theoretical aspects
and proof of concept. Journal of Forensic
Sciences 48: 593-603.
J.H. Wagner, G.M. Miskelly. (2003).
Background
correction
in
forensic
photography II. Photography of blood under
conditions of non-uniform illumination or
variable substrate colour - Practical aspects
and limitations. Journal of Forensic Sciences
48: 604-613.
J. Wawryk, M. Odell. (2005). Fluorescent
identification of biological and other stains on
skin by the use of alternative light sources.
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296-301.
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alternate light source to detect semen.
Academic Emergency Medicine. 9: 1045-1048.
E. Camilleri, E. Silenieks, J. Henry. (2006).
Locating Saliva Stains using the Polilight and
SALIgAE Spray, Evidence Recovery and
Biology Analytical Groups. Forensic Science
Australia, Government of South Australia.
R. Gaensslen. (1983). Sourcebook in Forensic
Serology, Immunology, and Biochemistry.
Washington, DC, U.S. Department of Justice.
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ultraviolet techniques used in the detection of
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High-Intensity Quartz Arc Tubes in the
Detection of Body Secretions. Journal of
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K.A. Santucci, D.G. Nelson, K.K. McQuillen,
S.J. Duffy, J.G. Linakis. (1999). Wood's lamp
utility in the identification of semen. Pediatrics
104: 1342-1344.
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C.D. Garbin, E.J. MacDonald. (2006). The use
of an alternative light source to detect semen in
clinical forensic medical practice. Journal of
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Improving the effectiveness of fluorescence for
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Additional information and reprint requests:
Khoo Bee Ee
(Email: beekhoo@eng.usm.my)
School of Electrical and Electronic Engineering
Engineering Campus
Universiti Sains Malaysia
14300 Nibong Tebal, Penang, Malaysia
27
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
Development of Reagent Test Kit for the Enhancement of
Shoeprints at Crime Scene
Umi Kalthom Ahmada, Noorul Huda Abdul Jabara, Chong-Hooi Yewb, Noor Azmi Yusoffb
a
Department of Chemistry, Faculty of Science, Universiti Teknologi MalaysiaSkudai, Johor, Malaysia.
b
Forensic Laboratory, Royal Malaysia Police, 8 ½ mile Jalan Cheras, Cheras, Selangor, Malaysia
ABSTRACT: Footwear marks constitute important trace evidence found in major crime scenes. Latent
footwear marks require efficient treatments for possible shoe sole pattern identification in crime analysis. This
study consisted of two parts: enhancement of footwear prints and development of reagent test kit for the
enhancement of shoeprints at crime scene. Chemical enhancements were carried out on shoe soles subjected to
muddy and bloodstained areas. The shoeprints were formed on nine different matrices including porous and
non-porous surfaces. Eighteen reagents were tested for the enhancements of muddy and bloodstained shoeprints.
Reagents employed on muddy prints reacted with either metal ion, amino acid or other component while
reagents employed on bloodstained prints were sensitive to protein, peroxidise and amino acid. Among the
eighteen reagents, leucomalachite green and patent blue were chosen as the best reagents in enhancing
bloodstained shoeprints while potassium ferrocyanide and sudan black were the best enhancement technique for
muddy shoeprints. All four reagents were effective on both porous and non-porous surfaces. However, the
reagents were not applicable on dark background surfaces. These reagents were used for the test kit
development for crime scene work. The test kit was named as SPECK (Shoeprint Enhancement Chemical Kit)
and consisted of chemical reagents, instruction, safety precaution, and personal protective equipments such as
gloves and masks. The reagent test kit developed was found to be reliable for screening purposes, rapid to carry
out and specific as well as reasonably cost.
Keyword: shoeprint chemical enhancement, crime scene toolkit, forensic chemistry
Introduction
When one object makes physical contact with
another, it may leave some of its physical
characteristics on the recipient in the form of an
impression [1]. Marks and impressions evidence
can be generally defined as the signature (pattern)
of an object that is left in another object when the
two have come in contact with one another [2].
Footwear creates marks at the crime scene called as
shoeprint and can be extremely informative to the
forensic investigator. The sole of footwear picks up
various kinds of material as a person walks, and
this is readily transferred to other surfaces, creating
an impression that can reveal the pattern on the sole
[3].
Many footwear have soles with distinctive tread
patterns, whereas others are smooth. Footwear
evidence can be extremely valuable in associating
perpetrators of crimes with the crime scenes. There
may be shoeprints at and near the entry points to a
crime scene, at the scene, and at and near the exits.
In fact, there are many more shoeprints at and
around crime scenes than are ever discovered or
collected. It is reasonable to conclude that there are
more potential footwear marks at crime scenes than
there are fingerprint impressions.
Latent or unclear shoeprints are usually harder to
be detected by the naked eyes and are frequently
overlooked by law enforcement personnel.
Moreover, most of the footwear impression or
shoeprints evidences are formed on static and
permanent surfaces; it is therefore not possible for
crime scene officers to bring back the evidence.
Hence, a means of enhancing the footwear
impression must be undertaken. Bloodstained and
muddy shoeprints recovered at crime scenes could
be enhanced using chemical reagents specific for
the detection of either blood or mud. Thus, various
types of analysis on stained shoeprints must be
carried out to find out the best enhancement
method.
This study was therefore undertaken to develop a
reagent test kit for the enhancement of shoeprints at
crime scene. Chemical reagents that are specific for
detection of bloodstained and muddy shoeprints
were investigated for their effectiveness on various
matrices for the enhancement of latent shoeprints.
28
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
Experimental
Materials: In this study, nine materials were used
as surface matrices of shoeprints, which consisted
of five porous surfaces and four non-porous
surfaces (Table 1). Blood and mud were used as
stained area throughout the study. Mud was
obtained from an identified location around the
Faculty of Science, UTM while screened, blank
expired blood (group A+ and O+) in plastic
packages of 300 mL were obtained from Hospital
Sultanah Aminah (HSA), Johor Bahru and Hospital
Tengku Ampuan Rahimah (HTAR), Klang to
produce muddy and bloodstained shoeprints
respectively. A Nikon D60 L11 digital camera (1855 mm f/3.5-5.6G VR Japan) were used for the
image capture of both untreated and treated
shoeprints, and a ruler (inch and centimeter scale)
were used to indicate the scale size in the
photographic shots. A North Star size 4 patterned
shoe sole was used to form the stained shoeprints
on the matrices (Fig. 1).
Table 1: Nature of matrices used in this study
Type of Surface Matrix
A4 White Paper
Cardboard
Carpet
Food Wrapper-Porous
Food Wrapper-Non-Porous
Linoleum-Dark
Linoleum-Light
Plastic
Tiles
Nature of Matrix
Porous
Porous
Porous
Porous
Non-Porous
Non-Porous
Non-Porous
Non-Porous
Non-Porous
Fig. 1: Bottom shoe sole pattern Different views of North Star shoe used in the study
Chemicals: Chemicals used in this study comprised
of patent blue VF, 5-sulfosalicylic acid dihydrate,
leucomalachite green, and sudan black B (SigmaAldrich - Steinheim, Germany), and potassium
hexacyanoferrate (II) from QRëC, New Zealand.
All chemicals used were of analytical grade.
Solvents used included analytical grade glacial
acetic acid from HmbG Chemicals, hydrochloric
acid from J.T. Baker (Philipsburg, USA) and
ethanol from F.A.D Ltd (England, UK), 2N nitric
acid (QRec, New Zealand), 30% hydrogen
peroxide, and diethyl ether. Double distilled water
was prepared in the laboratory.
Procedure: Muddy shoeprints were enhanced using
nine different reagents which react with either
metal ion, amino acid or other components while
bloodstained shoeprints were treated with nine
other reagents which react with either peroxidase or
protein. The best enhancement reagents were
identified for each muddy and bloodstained
shoeprints. A crime scene toolkit was using
selected reagents tested. The toolkit was named and
came together with personal protective equipments
as well as user manuals.
Muddy shoeprints were enhanced using potassium
ferrocyanide and sudan black reagents. Two types
of solutions were prepared for shoeprint
enhancement using potassium ferrocyanide reagent.
Solution A was prepared by mixing hydrochloric
acid (10 mL) and ethanol (90 mL) and for solution
B, potassium ferrocyanide (5 g) was dissolved in
deionised distilled water (100 mL). The articles
bearing latent muddy shoeprints were sprayed with
solution A followed with solution B [4]. Sudan
black dye solution was prepared by dissolving
sudan black B (3 g) in ethanol (250 mL). The
29
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
matrices containing impression were sprayed with
this solution and washed with distilled water [4].
Leucomalachite green and patent blue reagents
were chosed to enhance bloodstained shoeprints.
Leucomalachite green staining solution was
prepared by mixing leucomalachite green (1 g)
with diethyl ether (70 mL), glacial acetic acid (0.5
mL) and 30% hydrogen peroxide (0.3 mL). The
matrices were sprayed until completely covered
and the matrices were allowed to air dry. The green
coloured development of impressions occured
within 10 seconds [5]. Patent Blue dye solution was
prepared by dissolving sulfosalicylic acid (20 g)
and patent blue VF (2 g) in distilled water (1 L),
resulting in a greenish-blue solution. The objects
were stained by spraying the item with the dye
solution for approximately 1 minute followed by
rinsing it with distilled water [6].
of the shoeprints were observed and recorded after
the enhancement. In order to ensure an objective
way of comparison, photographs were taken before
and after treatment [1]. Besides, the purpose of
taking a photograph of shoeprints is to record it for
possible future comparisons with a suspect shoe.
Therefore, a scale should be available in the
photograph [7].
Results and Discussion
The best chemical enhancements of bloodstained
and muddy shoeprint on various matrices are
shown in Table 2. Fig. 2 and Fig. 3 show enhanced
bloodstained shoeprints using leucomalachite green
and patent blue reagents respectively. While
enhanced muddy shoeprints using potassium
ferrocyanide and sudan black were shown in Fig. 4
and Fig. 5 respectively.
Photographs of the enhanced prints were taken as
the matrix dried up. The length, width, and patterns
Table 2: Results of stained shoeprint enhancement with four different reagents
Bloodstained Enhancement
Mud stained Enhancement
Reagents
Reagents
Matrices/Reagents
Leucomalachite
Patent blue
Potassium
Sudan black
green
ferrocyanide
A4 White Paper
+
x
x
x
Cardboard
++
+
+
+
Carpet
++
+
++
+
Food Wrapper-Porous
++
+
+
x
Food Wrapper-Non-Porous
++
++
++
+
Linoleum-Dark
x
+
++
Linoleum-Light
+
++
++
++
Plastic
++
++
x
+
Tiles
+
++
+
x
“-“ : Bad;
“+” : Fair;
“x” : Poor;
“++” : Good
As a presumptive test, leucomalachite green is
sensitive to the protein stain in the blood [8]. It
gave a very contrast enhancement with porous and
non-porous surface matrices. Subsequently patent
blue solution which gave a good enhancement on
non-porous matrices was also chosen as a
complementary test to leucomalachite green
reagent. Potassium ferrocyanide which reacts with
metal ion in the soil [9] enhanced shoeprints on
almost all matrices and the resulting colour of the
stain was clearly visible. While sudan black stains
other component of the soil which is fat [10]. A
deep blue-black colour produced successfully
enhanced shoeprint on non-porous matrices.
All the selected reagents were kept in room
temperature for two weeks. After two weeks, the
reagents were tested again and the reagents still
gave positive results. Only leucomalachite green
was kept in the refrigerator because hydrogen
peroxide was used during the preparation of the
reagent. Hydrogen peroxide was stored in the
refrigerator to avoid oxidation of the reagents.
Thus, the reagents selected were used to develop
the reagent test kit named SPECK (Shoeprint
Enhancement Chemical Kit). The kit (Fig. 6)
consisted of seven reagent bottles. The entire
reagent bottles were labelled accordingly (Table 3).
The test kit was also equipped with personal
protective equipment like gloves and goggles, usermanual and notes regarding procedure of
enhancement. Goggles were included in the kit and
to be used for preventing the contact of reagent
with eyes upon spraying the shoeprints. Gloves are
used to avoid direct contact of hand with either the
shoeprint or reagents. Mask should be worn during
the enhancement to avoid the inhalation of the
reagents. The kit can be placed in room
temperature except for leucomalachite green, it is
advised to keep the reagent in the refrigerator. It
could be taken out and placed in a cooler box just
30
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
before the investigators depart to the crime scene.
Based on the chemicals and other apparatus to be
included in the kit, the kit was estimated to cost
around RM 400.00. This constituted a reasonably
cost kit.
Fig. 2: The bloodstained shoeprint mark on tiles before
(Top) and after (Bottom) treatment with leucomalachite
green
Fig. 3: The bloodstained shoeprint mark on non-porous
food wrapper before (Top) and after (Bottom) treatment
with patent blue
Fig. 4: The mud stained shoeprint mark on non-porous
food wrapper after treatment with potassium
ferrocyanide
Fig. 5: The mud stained shoeprint mark on linoleum
before (Top) and after (Bottom) treatment with sudan
black
31
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No.1
Table 3: Reagents and labels
No.
1
2
3
4
5
6
7
Reagents
Leucomalachite green
Patent blue
Deionised water
Mixture of HCl and ethanol
Potassium ferrocyanate
Sudan Black
Deionised Water
B1. Reagent A and B could be used on both porous
and non-porous surfaces.
Labels
A
B1
B2
C1
C2
D1
D2
Reagent C and D are to be applied on mud stained
shoeprints. For reagent C, the user should spray the
shoeprint with reagent C1 and followed with C2
until a blue colour is observed. For reagent D, D1
must be mixed with D2 before it could be sprayed
onto the impression. A blue-black colour will
appear upon spraying.
The reagent test kit developed was found to be
reliable for screening purposes, rapid to carry out
and specific as well as reasonably price.
However, the reagents are only applicable on light
coloured surfaces and not suitable to be applied on
dark coloured surfaces since all the enhancement
colour of the reagents were not visible on dark
surfaces. Moreover, the reagents could only
enhance blur or unclear latent shoeprints, not latent
shoeprint.
Acknowledgements
Thanks are due to the Department of Chemistry,
Faculty of Science, Universiti Teknologi Malaysia,
and Forensic Laboratory, Royal Malaysia Police,
Cheras, for research facilities. Special thanks and
appreciation also go to all the hospitals staffs: Mr.
Ariffin Hj. Mohd Yusof (Hospital Tengku Ampuan
Rahimah, Klang) and Dr. Zanariah Kassim
(Hospital Sultanah Aminah, JB) for their
assistances in obtaining blank expired screened
blood samples.
References
Fig. 6: Shoeprint Enhancement Chemical Kit showing
inside (Top) and outside (Bottom) view
For bloodstained shoeprint enhancement, reagent A
or B1 and B2 could be applied. Reagent A can be
straight away used to spray the shoeprint until a
bluish-green colour is observed. While for reagent
B, the shoeprint should be sprayed with B1 first
and washed with reagent B2. The shoeprint will
change to blue colour upon spraying with reagent
1. Dwane, S.H. (1995). The Art and Science of
Criminal Investigation; Footwear, the Missed
Evidence. The Lightning Powder Co.
Newsletter. 11: 2-5.
2. Sullivan III, W. T. (2007). Crime Scene
Analysis-Practical Procedure and Technique.
New Jersey: Pearson Prentice Hall.
3. Bodziak, W.J. (2000). Footwear Impression
Evidence. 2nd ed. Boca Raton: CRC Press.
4. Theeuwen, A.B.E., Barneveld, S. V., Drok, J.
W., Keereweer, I., Lesger, B., Limborgh, J. C.
M., Naber, W.M., Schrok, R. and Velders, T.
(2001). Enhancement of Muddy Footwear
Impression. Forensic Science International.
119: 57-67.
5. Theeuwen, A.B.E., Barneveld, S. V., Drok, J.
W., Keereweer, I., Lesger, B., Limborgh, J. C.
M., Naber, W. M., Schrok, R. and Velders, T.
(1998). Enhancement of Footwear Impressions
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6.
7.
8.
9.
in Blood. Forensic Science International. 133151.
Saferstein, R. (2007). Criminalistics: An
Introduction to Forensic Science. 7th ed. New
Jersey: Prentice Hall, Engelwood Cliffs.
Kramer, R. E. (2009). Footwear Impression
Photography. International Association for
Identification, Detective Cedar Falls Police
Department.
Ishihama, K., Koizumi, H., Wada, T., Iida, S.,
Tanaka, S., Yamanishi, T., Enomoto, A., Kogo,
M. (2009), Evidence of Aerosolised Floating
Blood Mist during Oral Surgery. Elsevier. 71:
359-364.
Yu, X. Z., Gu, J. D., Li, T. P. (2008).
Availability of Ferrocyanide and Ferricyanide
Complexes as a Nitrogen Source to
Cyanogenic
Plants.
http://www.ncbi.nlm.nih.gov/pubmed/1818086
2, retrieved on 30 April 2009.
10. Sigma Aldrich Catalog Entry for Sudan Black.
http://www.sigmaaldrich.com/catalog/search/P
roductDetail/ ALDRICH/860336, retrieved on
August 2009.
Additional information and reprint requests:
Umi Kalthom bt Ahmad
(Email: umi@kimia.fs.utm.my)
Department of Chemistry
Faculty of Science
Universiti Teknologi Malaysia
81310 UTM Skudai, Johor, Malaysia
33
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Skull-Photo Superimposition: A Remedy to the Problem of
Unidentified Dead in Malaysia
P.T. Jayaprakasha, Bhupinder Singhb, Ridzuan Abd Aziz Mohd Yusopc, Hetty Susilawati
Asmunid
a
Forensic Science Programme, School of Health Sciences, Universiti Sains Malaysia, Kelantan, Malaysia
b
Department of Forensic Medicine, Hospital Pulau Penang
c
Department of Chemistry Malaysia, Terengganu
d
Faculty of Law, Universiti Malaya
ABSTRACT: Skull-photo superimposition continues to be a popular method for suggesting individual
identification of unidentified human remains. In spite of the popularity in many countries, this method is not in
regular practice in Malaysia. Computer aided Video Superimposition Device has been fabricated for the first
time in Universiti Sains Malaysia paving the way for skull based identification in Malaysia. In Malaysia, about
25% of the skulls preserved in Hospital Pulau Pinang and among the human remains buried or cremated in two
hospitals in Kuala Lumpur are found to pertain to cases of equivocal death and/or possible homicide. As of now,
DNA based identification has been resorted to in about 10% of the bodies buried or cremated in the hospitals in
Kuala Lumpur and the rest continue to remain unidentified. At present these cases remain as cold cases. The
record of missing individuals in PDRM, Malaysia indicates about one thousand of the missing individuals to
continue to be untraced every year. It is proposed that a) a skull photo superimposition facility be established in
Forensic Science Laboratory, Kuala Lumpur for regular use to identify the dead bodies where the cause of death
is equivocal since such identifications are known to bring to light motives in homicides and b) a Central
Identification Laboratory similar to VICTIM project of FBI be established in Forensic Science Laboratory,
Kuala Lumpur for suggesting possible identification of all unidentified remains utilizing the face photographs of
missing individuals placing Malaysia on par with the countries pioneering in human identification elsewhere.
Keywords: Forensic anthropology in Malaysia, skull-photo superimposition, identifying the unidentified dead,
missing individuals
Introduction
Skull-photo superimposition is an accepted
scientific method for suggesting individual
identification of unidentified human remains such
as recovery of skeletonized remains, decomposed
or dismembered bodies as found during routine
criminal investigation, mass disasters, bodies
subjected to shearing forces as in explosions, burnt
or charred remains [1-14]. This method relies on
the use of commonplace evidence, face
photographs of missing individuals for comparing
with the skulls recovered from human remains
which render its applicability both popular and
effective. Ever since the premier acceptance of
skull-photo superimposition based identification in
the court of law in England [1], this method had
gained legal acceptance in most of the countries [1,
2, 9, 15-21] including Malaysia [22, 23]. The use of
face photographs of missing individuals bestows on
this method the advantage of being effective in
situations where blood relatives of the suspected
dead individuals cannot be located due to lack of
tentative identity thereby hindering DNA based
identification.
Although
skull-photo
superimposition has acquired popularity in most of
the countries [1-32], yet this method is not
regularly practiced in Malaysia.
The problem of unidentified dead bodies has been
well documented in forensic science [10-14, 34-39]
and skull-photo superimposition has been indicated
as a possible solution by recent workers [11-14].
Unidentified dead bodies, including those in which
cause of death is equivocal, are not infrequent in
Malaysia. In the absence of facility for skull based
identification, these dead bodies continue to remain
unidentified and are being disposed off as such
which may lead to dead end in the investigation
process. Since it has been found that establishing
identity leads to unraveling the motive in cases of
homicides, it is proposed that introduction of
anthropological methods such as skull-photo
superimposition and photo-photo comparison in
Malaysia may provide lead to further investigate
some of the cases that otherwise remain as cold
cases.
34
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Relevance of Skull-Photo Superimposition as an
Identification Technique
The popularity of skull-photo superimposition as a
method for identifying the dead by the FBI in the
USA recorded its peak utility during the nineties
and this appeared to reflect the availability of the
necessary equipment and expertise coupled with
the awareness of the value in forensic science and
law enforcement communities; this utility
diminished as molecular approaches have
increasingly become available and anthropologists
are
optimistic
expecting
photographic
superimposition in forensic science to increase [40,
41]. In tune with the above optimism, most of the
European countries [9-14, 24] and those in the east
[31, 32] are continuing the utilization of skullphoto superimposition. Albeit the lack of
appropriate man power, shifting to molecular
techniques for human identification may appear
befitting the western countries where dental records
form the prime source for identification. However,
in countries like India where dental records are
seldom maintained, skull-photo superimposition
continues to be in regular practice from as early as
during 1960. As stated by Ubelaker [41], the first
author found a trend that recorded an increase in
the utility of skull-photo superimposition from 15
cases during the period from 1971-1975 to 412
cases during the period from 1996-2000 [33] to
align with the availability of man power and
equipments coupled with awareness on the part of
the investigating officers.
Lack of appropriate maintenance of dental record is
a common problem in India (project) and the south
East Asian countries [9, 38, 39] including Malaysia.
Furthermore, during the victim identification in
Thailand following tsunami, among the western
victims 85.5% were identified using dental records
and 0.4% using DNA prompting the observation
that DNA based identification encountered certain
practical problems relating to decomposition and
the time involved [39]. On the other hand, about 73%
of the locals among the Thai victims still remained
unidentified and their bodies were being exhumed
until 2006 [40], the major reasons being lack of
dental records and the high cost of applying DNA
techniques. Recently, researchers have stressed the
grounds for morphological analysis instead of
solely relying on DNA based identification [13, 41,
42]. Under these circumstances, for Malaysia and
other south East Asian countries, resorting to DNA
technology for identifying human remains would
not obviate the need for applying conventional
anthropological measures such as skull-photo
superimposition.
Reliability in Skull-Photo Superimposition
In a survey on superimposition and reconstruction,
Aulsebrook and Iscan [43] indicate that the
credibility of superimposition system was shown
using it on human remains collected by the
Smithsonian Institute by Ubelaker et al [45].
Scientific verification on the reliability of photo
superimposition method has been attempted by
many researchers [26, 32, 45-47]. Among the
above researches, the work of Austin-Smith and
Maples [46] who verified superimposition using
one skull with multiple photographs found a chance
of about ten per cent wrong match between a skull
and an unrelated photograph. However, AustinSmith and Maples [46] used a 12 inch TV monitor
and hence did not evaluate fitness in ‘life size’.
Furthermore, these authors did not apply
anthropological measurements from the skull for
bringing out the life size of the face photographs.
At present, cranio-facial morphanalysis, an
additional procedure suggested for enhancing
reliability [8, 33] is also being used for verifying
the reliability. Preliminary studies (unpublished) in
Universiti Sains Malaysia indicate an enhanced
reliability in skull photo superimposition. It has
been indicated that craniofacial superimposition
can be considered reliable provided multiple
criteria are used for comparison [9]. The best
measure of a success of a method being court
acceptance, it is seen that the superimposition
method had gained acceptance in the courts in
many parts of the world as a method for suggesting
identification. As had been stressed by many
authors, this method cannot generate ‘definite’
identity [1-9]. It is well to remember the instructive
guideline of Thomas Dwight that ‘absolute
certainty’ is not the objective in anthropological
identification [40]. Rather, the expert is required to
assist in establishing the identity and it would be
for the judge to confirm it. Thus, the court
acceptance of this method is an endorsement on its
reliability for application in the context of
identifying the dead.
Court Acceptance: International Scenario
Identification of human remains using skull-photo
superimposition method has gained acceptance as
evidence in the courts in England, Australia, Hong
Kong, Japan and Malta [9]. Hagemeier [16]
described the video superimposition based
identification of a skull by Helmer and Gruner
during 1976, which gained acceptance by the
Frankfurt Assize Court. Krogman and İşcan [3]
point out that the video superimposition based
identification established by İşcan was accepted as
evidence of positive identification in one case
35
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
during 1980. Nickerson et al. [18] cited Brown et
al. [19] wherein Australian Court has accepted
video superimposition as an identification tool.
McKenna et al. [20] reported the acceptance of
identification by superimposition in the High Court
of Hong-Kong. Other countries where courts are
accepting superimposition as evidence include
France [16] Uruguay in South America,
Switzerland [9] and India [1, 8, 17, 21]. The
first author had testified on skull-photo
superimposition in the courts of Tamil Nadu,
India in 211 instances during the ten year
period from 1994 to 2004.
study as it was inexpensive. It has to be
mentioned here that the experts who had been
researching on superimposition for application
purposes in the courts law from early eighties
till recent times [3-15, 18-21, 24-33] had been
using video-vision mixer for producing the
superimposition effect of high quality and of
desired types and mirror based image mixing
would not serve such purposes. Another
research on craniofacial analysis in Malaysia
related to developing craniofacial database for
Malaysia population, an aspect not related to
individual identification in forensics [50].
Skull-Photo Superimposition: Court Acceptance
in Malaysia
Skull-Photo
Malaysia
Acceptance of identification of human remains
using superimposition in the courts in Malaysia has
been mentioned in three instances, two in a
student’s project report [24] and one in Criminal
Law Journal [23]. In all these instances, the
superimposition is indicated to have been carried
out by Prof. Dr. Masatsugu Hashimoto, University
of Tokyo, Japan. However, scientific details on the
type of equipments, anthropological parameters
used or the laboratory where the superimposition
was carried out etc. are not available. On the other
hand, the available records indicate the paucity of
local expertise as well as equipments in Malaysia
for carrying out skull-photo superimposition and
that Malaysia had been depending on foreign
expertise for tendering evidence in courts law when
necessity for identifying by superimposition
method arose.
The Computer aided Video Superimposition
Device (CAVSID) (Fig. 1) fabricated in Universiti
Sains Malaysia is the first of its kind in Malaysia.
The device consists of two high resolution (600
pixel) CCD video cameras (Bosch) with Fuginon
TV Zoom lens (Manual: 1.2/12.5-7.5) enabling
zooming the images in focus locked state for
capturing high resolution images of the skull and
face photograph. A digital video vision mixer
(Panasonic-G-MX70E) that enables real time
capture of the images in analogue form is
connected to the cameras. The images are cast in a
32 inch LCD TV monitor for analyzing and
recoding the measurements with least error as well
as for assessing the match between the skull and
face photograph during positioning the skull or in
process of mixing and wiping the superimposed
images. A remote-controlled pan and tilt device
(CS Lilin, PIH-303 Model) to which the universal
skull clamp is fabricated to be held on stand is used
for manipulating the various movements of the
skull for achieving the desirable positioning in
correspondence with the posture of the face.
Control unit for the pan and tilt device (PIH-301-C
(24VAC/240VAC) that enables finer movements of
the skull and is positioned in a location from where
the various movements of the skull for achieving
the desirable positioning can be effected. A VCR
(Sony-SLV-C317PS) for recording the real time
analogue
images
generated
during
the
superimposition process. Alongside, a 22 inch LCD
computer monitor (Samsung) connected to
computer system provided with video capture
software (Compro) is used for capturing the
superimposed images both frame by frame or as
video strip.
Skull-Photo Superimposition: State of the
Art of Research in Malaysia
Pertaining to the application of skull-photo
superimposition in Malaysia, identification of
a charred skull based mainly on the recovery
of part of a ‘Chanel’ ear ring has been
reported and skull-photo superimposition has
been mentioned although superimposing
images or the details of the methodology have
not been provided [48]. An article on
superimposition
published
in
Malaysia
indicates the use of Furue’s method wherein
two mirrors are used for producing the
superimposition effect [49]. Furue’s method
has been stated as the choice for the above
Superimposition
Facility
in
36
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Fig. 1: Computer Aided Video Superimposition Device (CAVSID) fabricated in Universiti Sains Malaysia.
1. CCD video camera (Bosch) with Fuginon TV Zoom lens (Manual: 1.2/12.5-7.5); 2. Digital video vision mixer (Panasonic-G-MX70E); 3.
32 inch LCD TV Monitor; 4. Pan and tilt device (CS Lilin TM, PIH-303) with skull clamp; 5. Control unit for the pan and tilt device (PIH301-C (24VAC/240VAC); 6. VCR (SonyTM-SLV-C317PS); 7. Computer system with image grabbing software and 22 inch LCD monitor; 8.
Improvised stands.
The captured images are printed in black and white
using a laser printer (1200 pixels resolution). The
minimum distance between the skull and the video
camera can be 1.2 m, and a similar distance is
maintained between the “life size” face photograph
and the other video camera. Two improvised
vertical stands are used for affixing the enlarged
face photograph and, the servo assisted pan and tilt
device supporting the skull can be fitted into
another improvised vertical stand. These stands are
provided with soft dark blue velvet cloth
background for avoiding shadow and scattering of
light. Diffuse fluorescent light from two 40 w tube
lamps positioned 2.3 m in front of the skull at a
height of 0.75 m above the level of the skull is used
for illumination for the skull as well as the face
photograph.
The Problem of the
International Scenario
Unidentified
Dead:
In Milan, Italy, during the 14-year period from
1995 to 2008, a total of 454 cadavers were received
as unidentified out of which positive identification
could be achieved for 62%. The mean unidentified
dead per year was 32. Roughly 50% of the
unidentified bodies were identified within 3 days
[15]. The authors list the most frequently methods
used in decreasing order as follow: visual,
fingerprints, soft tissue personal descriptions,
information from respective consulates, forensic
anthropology, forensic odontology and DNA. In
Denmark, during the period from 1992-1996, a
total of 89 unidentified dead bodies were received
in the three forensic institutes and 79 of them
(88.76%) were subsequently identified; the
methods in decreasing order being: personal effects,
dental examination, multiple sources such as X-ray,
tattoos, fingerprints etc. [10]. In France, during a
six year period from 2003 to 2009, a total of 134
unidentified bodies were received of which 10.2%
remained unidentified after all efforts. These
authors indicate that about half of the bodies were
inhumed with identity that was not scientifically
proved [16].
The Problem of the
Malaysian Scenario
Unidentified
Dead:
In Malaysia the second author had preserved 20
unidentified skeleton remains with skulls in
appropriately dealt facilities anticipating the future
identification since the cause of death was either
equivocal or indicating homicide. Forensic science
graduates from USM now serving as science
officers in a few other hospitals have initiated
preservation of skulls from unidentified bodies
although comprehensive data for entire Malaysia is
unavailable. During the 10 years period, between
1999 and 2008, in two hospitals in Kuala Lumpur
(HKL and UMMC) the total number of
unidentified dead bodies that have been buried or
criminated was 411 and in about 116 of them cause
of death was either in determinable or indicating
homicide [23]. DNA based identification had been
attempted in 12 of the 411 cases. Most of these
37
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
bodies have been indicated as ‘fresh’ and there is
no record whether it meant identifiable appearance
of the face since bodies can be classified as fresh
even in the presence trauma etc. in the face that
may hinder visual identification. Records relating
to subsequent identifications, if any, among the
above 411 cases are not available [23].
While subsequent identification is higher in other
countries, in Malaysia the available data does not
indicate subsequent identification. Suffice is to say
that anthropological means for identification are
prevalently used in other countries [10-15, 24]
while such facilities have not been established in
Malaysia so far. The data on the missing
individuals available in PDRM, Malaysia indicates
that about thousand missing individuals continue to
remain untraced every year (Table 2).
Comparison of the data published from other
countries [10, 13, 15] indicates that the unidentified
remains and those remaining so are considerably
more in Malaysia (Table 1).
Table 1: Comparison of the annual mean among the unidentified dead and rate of identification
Country
Period of study
Milan, Italy [24]
Denmark [10]
France [15]
Malaysia [23]
14-year period from 1995 to 2008
5-year period 1992-1996
6-year period from 2003 to 2009
10-year period from 1999-2008
Unidentified bodies
(annual mean)
32
18
22
41
Percent identified
62%
88.76%
89.8%
---
Table 2: Data on missing individuals; traced and remaining untraced (PDRM, Malaysia)
Years (5)
2004-2008
Total number of persons
reported as missing
Male
Female
4,622
10,528
15,150
Total number persons
traced alive
Male
Female
2,633
6,645
9,278
The Need for a Victim Identification Center in
Malaysia
Based on the available statistics, it may be
premature to conclude that all the cases indicated as
unidentified continue to be so till now. It is
possible that some of these cases had been solved
in the intervening period. It is necessary to have a
body for monitoring the progress in the
identification process so that the actual state of
these cases would be brought on record. In spite of
the above allowance for the statistics, it is fairly
evident all the bodies had been buried/cremated as
unidentified occur more frequently and in about 28%
of these bodies the cause of death was equivocal. It
is also evident that compared to the other countries,
the rate of identifying the dead in Malaysia is
considerably low. On the other hand, many face
photographs of missing individuals are being
routinely publicized in the media every day in the
hope that some of them may be recognized by the
public leading to identification of the missing
individuals, if he/she has been seen alive.
It is a fact that new photographs of missing persons
are being publicized every day and those that have
not been traced remain archived without much
scope for tracing. The above two ventures, first,
trying to establish identity of the unidentified
Total number of persons that
remained missing
Male
Female
1,989
3,883
5,873
remains, and, second, attempting to trace those
missing individuals utilizing media publication of
their face photographs, are both procedures that
presently remain dichotomous in Malaysia. As such,
there is scope in Malaysia for envisaging a
scientific project similar to the VICTIM project in
FBI for identifying the dead using anthropological
methods that would include skull-photo
superimposition and photo-photo comparison so as
to relate the skulls or the photographs of the dead
bodies with the archived face photographs of the
missing individuals for bringing out possible
identification, if any.
The project proposed here envisages i) collection of
face photographs of all the unidentified dead taken
by the investigation officers and ii) preservation of
skulls from all the unidentified human remains
where there is no lead on identity and forwarding
them to the Central Identification Laboratory that
would be a part of the Forensic science Laboratory.
All the face photographs of those missing
individuals who remain untraced would also be
forwarded to the Central Identification Laboratory
with details on personal particulars. Two major
identification ventures in the Central Identification
Laboratory would be a) to compare the skulls
retrieved from the dead with the face photographs
of the missing individuals taken while alive and b)
38
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
to compare the photographs of the dead with the
photographs of the missing individual taken while
alive. The outcome will be suggesting possible
identification. Identification so suggested can be
further followed up for DNA based identification
wherever
possible.
Standard
procedure
recommended for photo-photo comparison [51]
will be used for studying the photographs and a
Computer Aided Skull-photo Superimposition
Device (CAVSID) similar to the one recently
fabricated in the Universiti Sains Malaysia will be
used for suggesting skull based identification. Once
the identity is suggested through a scientific
method, it will provide a strong investigative lead
that may further unravel a motive and thus bring to
light some of the homicides that currently remain
buried without any recourse. In case of
implementing this project, Malaysia would prove a
pioneer in the South East Asia in utilizing
anthropological methods for solving the problem of
the unidentified dead.
Conclusion
Unidentified dead bodies are posing a challenge
that has been well reckoned internationally. In the
context of lack of dental records or DNA samples
from siblings, the only other option would be resort
to the conventional anthropological methods like
skull-photo superimposition. Lack of facilities for
skull-photo superimposition in Malaysia may prove
a void in the identification process. In the absence
of a method for skull based identification,
unidentifiable human remains would continue to
remain without identification leading to dead end in
the investigation process. Furthermore, this method
has potential use in utilizing the photographs of
missing individuals for suggesting possible identity
using skulls recovered from unidentified dead
bodies in routine identification of the unidentified
dead in ventures similar to victim identification in
FBI as well as in mass disaster victim identification.
Describing
the
Computer
Aided
Video
Superimposition Device fabricated for the first time
in Malaysia in Universiti Sains Malaysia, it is
proposed that skull-photo superimposition be
introduced and popularized in Malaysia alleviating
the problem of the unidentified dead in Malaysia.
Acknowledgements
The authors thank Universiti Sains Malaysia for the
financial
support
through
RU
grant
1001/PPSK/813011 that enabled fabricating the
Computer Aided Video Superimposition Device for
the first time in Malaysia. Prof. Syed Mohsin Syed
Sahil Jamalullail, Dean, Biomedical and Health
Science Research Platform, Prof. Ahmad Hj.
Zakaria, Dean, School of Health Sciences, and Prof.
Zainul F. Zainuddin, Innovation Office of
Universiti Sains Malaysia are thanked for their
constant encouragement and support during the
project for fabricating the superimposition device.
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Additional information and reprint requests:
P.T Jayaprakash
(Email: ptjaya@kb.usm.my)
Forensic Science Programme
School of Health Sciences
Universiti Sains Malaysia
16150 Kubang Kerian, Kelantan, Malaysia
41
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
A Review on Solid Phase Microextraction and Its Applications in
Gunshot Residue Analysis
Kah-Haw Changa, Ahmad Fahmi Lim Abdullaha
a
Forensic Science Programme, School of Health Sciences,Universiti Sains Malaysia, Kelantan, Malaysia
ABSTRACT: Sample preparation is important to prepare a sample for optimum performance characteristics
during analytical procedure. A review of papers on the practical applications of solid phase microextraction
(SPME) is presented particularly in the analysis of gunshot residue (GSR). The general introduction on SPME
and its theory are included. This review highlights the variables related to this technique during the analysis of
GSR in previous studies, namely type of SPME fiber used, sampling placement, adsorption and desorption
temperature and time, as well as the analytical techniques for final analysis. Apart from that, advantages and
limitations of SPME based gas chromatographic methods of GSR are explored. SPME is found to be a superior
sample preparation technique targeting for organic components, including GSR analysis. Gas chromatography
combined with varied detection systems performed prior to SPME gives low detection limits in simpler, easier
and rapid manner.
Keywords: solid phase microextraction, gunshot residue, firearm, sample preparation, gas chromatography
Introduction
In every analysis, multistep process is always
involved, including sampling, preparation of
sample, separation, qualitative and quantitative
determination, and statistical evaluation and finally
the result, decision and action. During any
analytical procedure, sample preparation is the
crucial step in order to prepare the sample for
optimum performance characteristics on its
examination. The recovery of the concentration of
components in interest is aimed to increase in
quantity apart from the elimination of any
interfering substance that may exist in a particular
sample. The efficiency and selectivity of sample
preparation technique undertaken during analysis,
as well as its applicability to the interested
compounds and matrices provide benefits in
numerous analytical techniques for good outcomes
and results. Additionally, easy to use, inexpensive
and compatible with a number of instrumental
methods are the requirements of an ideal sample
preparation technique. In this paper, we review the
solid phase microextraction (SPME) technique and
its forensic application particularly in gunshot
residue (GSR) analysis.
SPME Technique
Various techniques for sample preparation have
been developed previously. It was difficult to
extract the total quantity of compounds by both dry
and wet methods in the past for the purpose of
determination and quantitation. Extraction of a
sample with different solvent, evaporation steps
with gas phase collection, solid phase extraction
(SPE) and most recently SPME have been carried
out in published studies. SPME is a sample
preparation or extraction technique developed by
Pawliszyn and his group in early 1990s which can
be used in both in laboratory and on-site [1, 2].
This technique has gained popularity among
scientists and researchers with the presence of
advantages in its application in various fields,
including forensic perspective.
In SPME technique, it involves the use of fused
silica fiber, which is very well defined in
cylindrical surface geometry. This configuration
allows easy assessment of analyte to and from the
surface. This fiber is coated with an extracting
phase. Such extracting phase can be a liquid or a
solid comprised of a variety of conventional
stationary phases. Different kinds of analytes,
including both volatiles and non-volatiles can be
extracted from different kinds of media in either
liquid or gas phase. With SPME, analytes are
trapped on the fiber coated with liquid polymer or
solid sorbent during extraction followed by
desorption into the chromatographic mobile phase
for analysis [1, 2].
SPME is an equilibrium method that incorporates
sampling, isolation and enrichment into a single
step, applicable to gaseous, liquid and solid
samples. It is a simple, convenient, reproducible
and sensitive means for extraction of forensic
specimen with wide linearity range and relatively
good precision. SPME technique can be used for
the extraction of analytes from very small samples
due to its setup. Lower detection limits are
permitted by its application at part per trillion
42
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
concentrations. The elimination of solvents,
reduction of blanks and also the extraction time to a
few minutes are advantages of this extraction
technique, reducing the cost and the risk of
exposure to potentially hazardous solvents. As an
equilibrium technique, it eliminates any exhaustive
extraction of a sample. Additionally, the chemical
equilibria in a particular system is not likely to be
disturbed or disrupted as only a minute amounts of
analytes are removed on extraction. Additionally, it
allows multiple sampling and preservation of the
sample while minimizing the risk of contamination
due to sample handling and storage [1-4].
equilibrium between sample matrix and fiber
coating is achieved. With the achievement of
equilibrium, the amount of analytes extracted by a
SPME fiber is proportional to its concentration in
the sample without being dependent on the sample
volume. Such relationship gives the basis for
quantitation on analytes that are present in sample.
For both direct and headspace system, the
equilibrium concentration is independent on the
fiber location. Volatile analytes are extracted faster
than those which are semi-volatiles due to their
higher concentration in the headspace system,
contributing to faster mass transport rate [1, 2].
A number of fiber types have been considered
previously in SPME technique to maximize its
potential applications such as polyacrylate (PA),
carboxen, polydimethylsiloxane (PDMS) and
divinylbenzene (DVB) as the commonly used types.
Generally, the types of fiber to be considered
depend on the compounds that desired to be
extracted from a sample. The chemistry of
compounds may decide the adsorption and
desorption behaviours on a particular fiber type.
Analytes varied in polarities may require different
fiber chemistries. In conjunction, the thickness or
area of each fiber can be differed which decides the
sensitivity of the fiber. The temperature has a
significant effect on the kinetics of this extraction
process which depends highly on the thermal
stability and volatility of the compound interested
for analytical measurement. Apart from that, the
optimum extraction times of this technique are
affected by the combination of variables that
mentioned previously [1-4].
After extraction, the fiber is withdrawn into the
needle of SPME device and transferred to the
injection port of instrument for desorption of
analytes to be taken place. This is carried out with
the piercing of the needle through a septum.
Therefore, analytes are desorbed thermally and
transferred with the carrier gas into the column in
high temperature condition. Before any analysis, it
is crucial for blank runs to be performed between
samples in order to ensure that the fiber and GC
injection port were free of residues. Moreover, the
calibration of SPME requires special care which
generally consists of proper standard mixture
preparation, proper calibration and establishment of
the relationship between instrument response and
analyte content. Finally, different detector systems
can be considered for the identification and
quantitative analysis, depending on the required
sensitivity and selectivity of the performed analysis
[1, 2].
Using SPME, there are two sampling placement of
fiber for extractions that can be performed, namely
direct extraction and headspace (indirect)
configuration. Direct extraction mode involves the
direct partitioning of analytes from the sample
matrix to the extracting phase. Therefore, the
coated fiber of SPME is inserted directly into the
sample being examined. In latter mode, air
becomes the barrier for extraction. Analytes need to
be transported through the barrier to reach the
coating. However, this placement of fiber protects
the coating of fiber from damage by non volatile
and high molecular weight interferences that could
be present in sample matrix. Generally, more
volatile and more hydrophilic analytes are said to
be best recovered by headspace sampling. On the
other hand, direct sampling from aqueous samples
is the preferred mode for less volatile and more
hydrophobic compounds [1, 2].
Forensic Application in GSR Analysis
As the coated fiber placed in contact with the
sample, the transport of analytes into the coating
from the matrix starts. Generally, the extraction is
considered to be complete when the distribution
Since the development of SPME as a powerful
sample preparation technique, it has been used
extensively in the analysis of chemical traces in
forensic applications, including toxicology [5, 6],
narcotics [7-11], arson investigation [3, 12-17] and
explosive identification [3, 18-23]. Here, we review
the previously published studies on the application
of SPME technique in GSR analysis. The variables
in such application, namely the fiber chemistry
(type and thickness of stationary phase), sampling
placement, adsorption and desorption temperature
and time, as well as the analytical techniques for
final analysis are included. Additionally,
advantages of SPME in GSR analysis and its
problems encountered during the analysis are also
mentioned in this review.
The application of SPME technique in GSR
analysis has gained popularity in recent years. In
such analysis, volatile organic compounds (VOCs)
from the gunpowder or propellant composition of
ammunition are the targeted analytes to be
43
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
examined. VOCs refer to the organic compounds
with significant high vapour pressure at
atmospheric condition. In other words, they tend to
vapourize in atmospheric pressure and temperature.
Propellant, a chemical of mixture of chemicals,
found inside the cartridge case is the fuel used to
fire a bullet. Burning of the propellant initiated by
the ignition of primer produces gases rapidly
followed by the sudden increase of pressure inside
the case. As a result, the bullet is detached from
casing and directed towards the target [24]. The
formula of the gunpowder could be varied based on
the use of particular ammunition due to the desired
ballistics performance and stability characteristics.
However, the analysis of VOCs was found to be
difficult due to their low concentration in a sample.
In addition, analytical standard and non-polluted
sample preparation are required during their
examination. Therefore, SPME is found to be the
solution for these problems.
In 1998, SPME was first employed by Andrasko et
al. in the examination of GSR [25]. In the
performed study, the time since discharge was
estimated through SPME sampling from the
atmosphere inside the barrel of a shotgun. The
detector systems, gas chromatography/thermal
energy
analyzer
(GC/TEA)
and
gas
chromatography/flame ionization detector (GC/FID)
were used for its determination on the basis of the
rate of escape of volatile combustion product as a
function of time. With SPME technique, the
analysis of GSR on the estimation of time since
discharge has continued on the different types of
firearms and ammunition. Following the study by
Andrasko et al. (1998), the SPME technique has
been used successfully for detection of various
volatile compounds inside firearm barrels [26-28]
as well as inside spent cartridges [29-31]. This
SPME is said to be suited for headspace sampling
from narrow spaces like firearm barrel by simply
pushing the SPME holder inside the barrel from the
muzzle of a firearm [25-27]. As compared to other
headspace sampling techniques, passive adsorption
process of SPME has minimized the disturbance of
such extraction to the system.
For the improvement of SPME technique,
modification of the liner on the injector of a
chromatographic system was suggested by
Andrasko and Stahling (2000) and Andrasko and
Stahling (2003) [26, 27]. According to the authors,
specially designed liner for desorption of VOCs
was used in the injector compartment of a GC
instrument. Such liner with smaller inner diameter
(0.75 mm) reduces the desorption volume inside
the liner and thus minimize the peak broadening of
most VOCs. Apart from that, the increment in the
sampling time using SPME was found to improve
the sensitivity for detection of VOCs [27]. Other
than TEA and FID as the detection systems as
mentioned previously, mass spectrometry (MS) is
another technique that was considered for GSR
analysis following extraction procedure by SPME
which was greatly increased in its use for forensic
purposes [30-32].
Since the VOCs present in a cartridge tends to
disappear into the surroundings with time, a
procedure to preserve them from vaporization have
to be developed for analysis. In 2003, the method
of placing a shotgun shell in a vial was suggested
by Wilson et al. in order to prevent the escape of
VOCs before analysis [30]. In the method
suggested, a hole was drilled through the center of
the lid but not through the Teflon inner lining of
the cap. This allowed for easy SPME sampling
without opening the vial or disturbing the shell.
Additionally, repetitive samplings can be
performed easily without disturbing chemical
equilibria in the system. There was no observable
tendency for the repeatedly sampled cartridges to
show lower concentration [30]. However, this was
criticized by Weyermann et al. (2009) as repetitive
sampling may decrease the amount of analytes with
consecutive sampling, especially for smaller caliber
type ammunition [31].
In 2009, SPME was adopted to extract OGSRs
from a single particle of partially burnt gunpowder
by Burleson et al. (2009) [33]. OGSRs from only
one single particle of partially burnt gunpowder
were successfully extracted by SPME and analyzed
by gas chromatography/nitrogen phosphorus
detection (GC/NPD). The study showed that the
new extraction method is capable of extracting
trace amount of organic GSRs as signature
molecules for the identification of GSR. Recently,
the introduction of ion mobility spectrometry (IMS)
techniques following SPME permitted the detection
of trace amount of smokeless powders which are
the main components in ammunition, on the
discharge of any firearm. With the combination of
SPME and IMS, the detection limit in nanogram
range can be achieved in very short analysis time
[32]. A summary of published applications of
SPME on GSR analysis is presented in Table 1.
44
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Table 1: Published applications of SPME on GSR analysis
Source of
GSR
Shotgun
Sampling
placement
Headspace
(exposed to
atmospheri
c inside
barrel)
Fiber coating
PA, 85 µm
Rifle
PA, 85 µm
Pistol and
revolver
Headspace
(exposed to
atmospheri
c inside
barrel)
Carboxen/
PDMS, 85
µm
PA, 85 µm
CW/DVB, *
Spent
cartridge
Shotgun
shell
Spent
cartridge
NG, 2,4-DNT,
2,6-DNT,
unidentified
compounds
DPA, Nap
Headspace
(exposed to
atmospheri
c inside
barrel and
cartridge)
Headspace
(exposed to
atmospheri
c inside
cartridge)
Target analyte
Unidentified
compounds
Unidentified
compounds
NG, unidentified
compounds
PA, 85 µm
Naphthalene,
biphenyl
Headspace
(from
corked and
vial
samples
with shells)
PA, 85 µm
Naphthalene,
biphenylene,
DPA,
unidentified
peak (10.9 min)
Headspace
(from vial
samples)
PA, 85 µm
PDMS, 100
µm
Carboxen/
PDMS, 75
µm
Benzonitrile,
phenol, 2-ethyl1-hexanol,
naphthalene,
1,2dicyanobenzene,
DPA
Smokeless
powder
(Commerc
ially
available)
Headspace
(from vial
samples)
Gunpowd
er particle
Headspace
(from vial
samples)
PDMS, 100
µm
PDMS, 100
µm
DPA, EC, 2,4DNT, 2-ethyl-1hexanol
DPA, NDPA,
DNDPA, MC,
EC
Extraction and
desorption
conditions
TC= 200oC, C= 7
min, E= 30 min,
TS= r.t, TD=
170oC, D= 7 min
TC= 200oC, C= 7
min, E= 30 min,
TS= r.t, TD=
200oC, D= 7 min
TC= 200oC, C= 7
min, E= 20 min,
TS= r.t, TD=
170oC, D= *
TC= 170oC, C=
10 min, E= 40
min, TS= r.t, TD=
170oC, D= *
Limit of
detection
(LOD)
Peak
height ca.
500 mV
Peak area
500
Peak
height ca.
0.3-14.5
mV
(muzzle),
2.0-11.8
mV
(cartridge)
Peak
height ca.
0.35-8.6
mV
TC= 200oC, C= 7
min, E= 20 min,
TS= r.t, TD=
170oC, D= 7 min
TC= 200oC, C= 7
min, E= 20 min,
TS= r.t, TD=
170oC, D= 7 min
TC= 200oC, C=
10 min, E= 20
min, TS= r.t, TD=
200oC, D= 10
min
Peak
height ca.
0.5 mV
TC= 280oC, C=
15 min, E= 40
min (80 min),
TS= 40oC, TD=
280oC, D= 5 min
nanogram
TC= *, C= *, E= 5
– 120 min, TS= *,
TD= 280oC, D= *
TC= *, C= *, E= 5
– 40 min, TS= *,
TD= 215oC, D= *,
TDT= 180oC
TC= 250oC, C= 5
min, E= 45 min,
TS= 95oC, TD=
250oC, D= 2 min
0.15 – 3
ng
Peak area
ca. 17-23
Final
analysis
Reference
GC/TEA
[25, 28]
GC/FID
GC/TEA
[26]
GC/TEA
[27]
GC/TEA
[29]
GC/FID
*
0.12-1.2
ng
GC/MS
[30]
GC/MS
[31]
GC/MS
[32]
IMS
10 ng
GC/NPD
[33]
TC= Conditioning temperature, C= Conditioning time, E= Fiber exposition time, TS= Sampling temperature, TD= Desorption temperature,
D= Fiber desorption time, TDT= Drift tube temperature; PA= polyacrylate, PDMS= polydimethylsiloxane, CW= carbowax, DVB=
divinylbenzene; NG= nitroglycerine, DNT= dinitrotoluene, DPA= diphenylamine, NDPA= nitrodiphenylamine, DNDPA=
dinetrodiphenylamine, MC= methyl centralite, EC= ethyl centralite; GC= gas chromatography, TEA= thermal energy analyzer, FID= flame
ionization detection, MS= mass spectrometry, NPD= nitrogen phosphorus detection, IMS= ion mobility spectrometry; *= not given
45
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Conclusion
During analysis, the main difficulty encountered in
method developments was the reliability of the
sampling method. SPME was shown to be a
superior alternative for such purpose, including in
the analysis of GSR. In GSR analysis, desorption
and analysis of volatile samples collected by SPME
is most often performed using GC with different
detection systems. As mentioned by numerous
authors, SPME can give low detection limit with
simpler, easier, faster and sensitive procedure.
Considerably advantages do exist for SPME
followed by sensitive analytical analysis was found
to be adequate for analysis of organic volatile
residues, especially in GSR.
8.
9.
10.
Acknowledgements
Institute of Postgraduate Studies (IPS), Universiti
Sains Malaysia is thanked for financial support.
11.
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Additional information and reprint requests:
Chang Kah Haw
(E-mail: edwardchangkahhaw@hotmail.com)
Forensic Science Program
School of Health Sciences
Universiti Sains Malaysia
16150 Kubang Kerian, Kelantan, Malaysia
47
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Analysis of Gunshot Residue Deposited on Cotton Cloth Target at
Close Range Shooting Distances
Mohamed Izzharif A. Halima, Umi Kalthom Ahmada, Chong-Hooi Yewb, Muhammad Koey
Abdullahb
a
Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia, Skudai, Johor, Malaysia
b
Royal Malaysia Police Forensic Laboratory, Mile 8 ½, Cheras, Selangor, Malaysia
ABSTRACT: The study of gunshot residue (GSR) has become imperative in forensic firearm examination. It
may be used to estimate firing distance, to identify bullet holes and to determine whether or not a person has
discharged a firearm. This study emphasized on the examination of the pattern of GSR particles deposited on
cotton cloth target at different shooting distances (3 inch-12 inch). Two types of pistol (semiautomatic and
revolver) and ammunitions (Syarikat Malaysia Explosive (SME) and Winchester) were used for the shooting
test. All shooting tests were performed at Lapang Sasar of Pasukan Gerakan Am PDRM, Cheras. The results
showed that GSR particles produced on cotton cloth by semiautomatic pistol using SME 9 mm ammunition
generated higher amount and larger GSR particles which were almost completely burnt particles. The nitrite
residues were confirmed using modified Griess test that revealed semiautomatic pistol produced considerable
amount of nitrite residues. Scanning Electron Microscope (SEM) micrograph of GSR particles discharged from
semiautomatic pistol using SME 9 mm ammunition revealed a mean particle size of 2.6 µm with spherical
shapes. On the other hand, using revolver pistol with SME 0.38 Special ammunition, the GSR particles
produced were small in amount. The modified Griess test also revealed that revolver pistol generated tiny
amount of nitrite residues. Nevertheless, using Winchester 0.38 Special ammunition, the GSR particles
discharge were of high amount and large in size which composed of completely unburned, partially burnt and
completely burnt particles. Whereas SEM micrograph of GSR particles discharged from revolver pistol using
Winchester 0.38 Special ammunition generated irregular cluster grain with a mean diameter of 3.3 µm. Sodium
rhodizonate test was successfully applied for detecting the presence of lead residues around the bullet hole for
both pistols.
Keywords: gunshot residue, modified Griess test, scanning electron microscope, sodium rhodizonate test.
Introduction
During investigative proceedings of incidents
involving the use of firearms, forensic expertise
can provide key elements for the court to reach an
opinion. In such context, particular attention is
usually drawn to evaluative issues associated with
firearms and related evidence such as gunshot
residues (GSR). Upon arrival at the scene, it is the
responsibility of the police to start the difficult job
of determining the sequence of events of the case
especially when shooting is involved. When a gun
is being fired, both burned and unburned powders
from the propulsive charge as well as components
from the primer, the bullet and the cartridge case
form a cloud that may cause a roughly circular
pattern around the bullet hole upon contact which
is called GSR [1].
The pattern of GSR is significantly influenced by
various factors including the different muzzle-totarget angles, firing distances, type of ammunition
and weapon parameters, caliber and barrel length
[2]. However, there is increasing pressure from the
judicial system to identify an ammunition type, and
hence the weapon type which may have been
involved in an incident based on GSR collected [3].
Thus, this study emphasize on the pattern of GSR
on cloth target for different pistols and ammunition
used.
It is apparent that collecting cloth evidence bearing
GSR has more potential for providing inculpatory
evidence in some cases than bare hand sampling.
GSR retained on clothing are considerably longer
than skin surfaces resulting in a high probability of
their detection and identification from that source
[4]. If someone is standing close enough to the gun
as it is fired, the GSR will appear on the person’s
clothing. Thus, the estimation of distance from the
weapon’s muzzle to target material of clothing may
be in an important factor in an investigation [5].
The pattern of gunshot residue may vary with
different distances [6]. Numerous studies have been
published on the analysis of GSR by various
analytical methods. At close distances when many
smokeless powder particles are present on target
48
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
material, a typical pattern can be established by the
modified Griess test and hence the shooting
distance can easily be estimated [7]. Another
chemical test used to estimate the firing distance is
known as sodium rhodizonate test which is used as
a spot test for lead residues [8]. Therefore, these
chemical tests were used in this study to restore
insufficient interpretation of GSR pattern from
visual examination by the naked eyes as well as
microscopy.
at different distances from the shooter. Two types
of firearm, a 0.38 Revolver Smith & Wesson and a
9 mm semi-automatic Yavuz 16 Compact were
prepared. Prior to firing on the cloth target, at least
three shots were fired to safe target area in order to
ensure that the barrels will be coated with smoke
and powder residues for the whole series of shots to
the target samples. The set-up for this experiment
and all shooting were performed by specialized
police officers at Lapang Sasar Pasukan Gerakan
Am (PGA) PDRM, Cheras.
Experimental
Shooting Test
Chemicals and Materials
The objective in this shooting test was to estimate
the effective firing distance from the muzzle to the
cloth target. The distances were focused at close
ranges which were 3, 6, 9 and 12 inches. These
distances were chosen in this shooting test as the
nitrite is only effectively detected at the distance
less than 15 inch (Jeffrey, 1998). The shots were
tested at 90 degrees to the target. Each of the
distance was repeated three times so that the
modified griess test (MGT) and sodium
rhodizonate test (SRT) could be applied on separate
cloth target bearing the GSR.
The chemical reagents needed for the chemical
tests were sodium rhodizonate, sodium bitartrate
from Sigma Aldrich, Germany; tartaric acid,
sodium nitrite and methanol from QReC;
hydrochloric acid from J. T. Baker, Mexico; and
sulfanilic acid, alpha-naphthol and glacial acetic
acid from Merck from Germany. The chemicals
were all of analytical grade from a range of
suppliers. Two types of pistol used for the shooting
test were a 0.38 Revolver Smith & Wesson (serial
number: D943607 from USA) and a 9 mm semiautomatic Yavuz 16 Compact (serial number: T
0624-06 TF 022891 from Russia). The ammunition
used were Syarikat Malaysia Explosive (SME) 9
mm with full metal jacketed (FMJ) round nose
design, SME 0.38 Special with lead round nose
(LRN) design and Winchester 0.38 Special with
lead round nose (LRN) design. In order to perform
chemical tests, uncontaminated iron (Panasonic
brand), plastic tray, spray bottle (300 mL), filter
paper (A4 size) and cotton swab were procured.
Other materials and apparatus (stand, soft plastic
cardboard pad, stick to hold the cloth item) relating
to the shooting tests were provided by PDRM. A
Nikon D60 18-55 VR Kit digital camera (made in
Japan) was used to photographed the GSR images.
Instrumentation
Scanning electron microscope (SEM) model FEI
Quanta 200F equipped with xT Microscope
Control software was employed for morphological
and microstructure of GSR particles.
Target Material and Firearm Preparation
Cotton cloth targets (25 cm x 25 cm) were used for
the shooting test. Each cloth was placed over a 60 x
65 cm soft plastic cardboard pad and stapled to it.
The soft plastic cardboard padding which was fixed
to a wooden frame stand was 115 cm above the
ground. The cloth was not stretched but pulled tight
enough to eliminate wrinkles in the fabric. The
stand bearing the cloth target was then positioned
Chemical Tests
For the modified Griess test, the precut sheets of
filter paper (A4 size) were briefly submerged in the
mixture consisted of a solution of 0.5 g of sulfanilic
acid in 100 mL of deionized water and a solution of
0.28 g of alpha-napthol in 100 mL of methanol.
Then, 15 % acetic acid was sprayed on the cloth
bearing GSR particles on the side of bullet entrance.
Then, the sample was placed face down on the
filter paper. Again, acetic acid was sprayed but on
the side of bullet exit until the cloth was dampened.
The layers were pressed with a hot uncontaminated
iron. Acetic acid steam was forced through the
layers, causing the colour-producing reaction. The
cloth item was then separated from the filter paper.
An orange colouration on the paper indicates the
presence of nitrite residues.
For sodium rhodizonate test, the test area was
initially sprayed with the saturated solution of
sodium rhodizonate. The solution was prepared by
dissolving a small amount of sodium rhodizonate
(about 0.2 g) in 100 ml deionized water to form a
saturated solution that was approximately the
colour of dark tea. After that, the same area was
then sprayed with the buffer solution. The buffer
solution was prepared by dissolving 1.9 g of
sodium bitartrate and 1.5 g of tartaric acid in 100
mL of deionized water. The resulting solution was
heated on a hot plate while agitating with a
magnetic stirrer to produce a pH 2.8 buffer solution.
This solution eliminates the yellow background
49
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
colour caused by the sodium rhodizonate,
establishes a pH 2.8, and displays a pink colour in
the presence of lead.
SEM analysis
Scanning electron microscopy (SEM) of individual
particle analysis was employed for this study to
examine the characteristic GSR particles. As the
GSR is non-conductive, low vacuum mode was
used. GSR were sampled from the cloth target by
carefully pressing the double-sided carbon base
tape cut to 8 mm in length onto cloth bearing GSR
particles. Subsequently these tapes were fixed to a
cylinder that was rotated within the SEM chamber
and adjustable in an axial direction. The tapes
bearing GSR particles were scanned for possible
GSR particles with simultaneous SE and X-ray
imaging. The outcome was a microstructure image
of the GSR particles with desired magnification.
Results and Discussion
(A) Semiautomatic Pistol
GSR at different shooting distances
1) Visual Examination
The estimation of shooting distance were first made
based on the direct and visual examination of the
cloth target and then treatment with specific
chemical regents as stated by proposed by previous
researchers [9]. Sodium rhodizonate test, a method
that would reveal the GSR pattern on the item in
situ can minimize the loss of the GSR during the
transfer of the evidence. Whereas, modified Griess
test needs to be performed in the laboratory as it
requires troublesome procedures. The shooting
distances selected were 3, 6, 9, and 12 inches. After
the cloth target was shot, the pattern of the GSR
produced was photographed using a Nikon digital
camera before subjected to further chemical tests.
There were three areas in the GSR pattern that need
to be observed and examined which were the
plume line, particle and sooting of GSR. These
three areas could serve as the indicator for the
characteristic of the pattern of GSR at different
shooting distance. Fig. 1 shows the appearances of
cotton cloth shot using semiautomatic and revolver
pistols respectively at different shooting distances.
2) Modified Griess Test
Therefore, the modified Griess test is a test to
detect the presence of nitrites residues. After
visually examine an exhibit of a real case, modified
Griess test could be applied to determine a muzzleto-garment distance. The colour of orange specks
on the filter paper were the result of chemical
reaction involved between acetic acid vapors
(steam ironing on the back of cloth target), nitrite
residues and the chemicals contained in the filter
paper [9]. For cloth target shot using semiautomatic
at different shooting distances, intensity of the
orange colouration decreased with increasing
distance of muzzle-to-target. This implied that
nitrite residues could still be detected up to 12 inch
shot distance. It is estimated beyond 15 inch
muzzle-to-target distance, nitrite residue could not
be detected on the cloth bearing GSR fired by both
types of pistols. This is in agreement with that
reported by previous study [10].
Another test conducted on the cloth bearing GSR
was the sodium rhodizonate test. Information from
this test may also assist in establishing a muzzle-totarget distance. The positive test for sodium
rhodizonate test indicates that the lead may
originate from lead-bearing bullet or/and vaporous
lead residue from the primer. The test was
performed by spraying the exhibit with a weak
solution of sodium rhodizonate followed by a
buffer solution which caused the background
colour to disappear. The reagent reacted with any
lead that may be present and produced very bright
pink colour. At a firing distance of 3 inch, an
intense pink circular colouration around the bullet
hole was clearly visible on cloth target fired using
semiautomatic pistol. With increasing firing
distance from 3 to 12 inch, the pink circular
colouration appeared to be less intense. For
distance of 12 inches, the pink colouration was
very tiny indicating that the lead residue could only
be detected up to 12 inch away from the cloth
target. The diameter of the pink colouration
gradually decreased and the pink colouration
became less intense with increasing distance.
Therefore, it indicated that the lead residues were
only present in considerable amount at very close
firing distance.
3) Sodium Rhodizonate Test
It is found that the particles appeared as spherical
shape with an average diameter 1.5 µm. Some of
them were perfect spheres, sometimes hollow, and
minor were irregular and distorted which is in close
agreement with previous study [11].
When a firearm is discharged, nitrite particles are
expelled from the muzzle of a firearm and can be
imbedded in or deposited on the surface of a target.
Nitrite residues are by-products of completely
burnt particles of smokeless gunpowder.
50
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
(A)
collected from the cloth using double-backed
carbon tape SEM stub. It was then placed on the
SEM stage for further examination. By means of
computer system, the stub was scanned to locate
specific area that could produce an image of the
GSR particles either by low or high magnification.
Fig. 2 represents SEM micrograph for a GSR
particle discharged from semiautomatic pistol using
SME 9 mm ammunition.
(B)
(C)
Fig. 2: SEM micrograph for GSR particles discharged
from semiautomatic pistol using SME 9 mm ammunition
at 8000x magnification.
(B) Revolver Pistol
GSR at different shooting distances
1) Visual Examination
(D)
In this study, the pattern of GSR deposited on cloth
target for revolver pistol was also investigated. All
the methods and procedures involved when dealing
with semiautomatic pistol were repeated for the
revolver pistol. Therefore the examination and
analysis for the GSR produced using revolver pistol
were based on semiautomatic pistol in order to
estimate the shooting distance. Fig. 3 shows the
GSR pattern generated on cotton cloth target using
revolver pistol.
2) Modified Griess Test
Fig. 1: Pattern of GSR on cotton cloth fired using
semiautomatic pistol at different firing distances (A) 3
inch, (B) 6 inch, (C) 9 inch and (D) 12 inch.
Scanning Electron Microscopy
Scanning electron microscope was carried out to
determine the morphology and microstructure of
the GSR particles. For the examination, GSR were
After visual examination, the cloth target bearing
GSR were subjected to the modified Griess test to
visualize the nitrite residues expelled from the
revolver pistol. As in modified Griess test for
semiautomatic pistol, the colour of orange specks
on the filter paper appeared as a result of chemical
reaction involved. As expected, the intensity of the
orange colouration decreased with increasing
distance of muzzle-to-target. However, the orange
specks appeared on the filter paper were not too
51
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
intense and crowded. It shows that the revolver
pistol generated little amount of nitrite residues.
Yet, it is believed that the nitrite residues could still
be detected up to 15 inch because there was still
nitrite residues observed on the filter paper at
distance of 12 inch.
(A)
3) Sodium Rhodizonate Test
The subsequent chemical test treated was sodium
rhodizonate test. The cotton cloth target bearing
GSR expelled from revolver pistol was sprayed
with sodium rhodizonate solution and buffer
solution to observe the appearance of pink colour
as a result of chemical reaction. The diameter of the
pink colouration gradually decreased and the pink
colouration became less intense with increasing
distance. However, the pink pattern was difficult to
visualize beyond 9 inch. Results of this study
indicated that lead residues from vaporous lead
smoke (primer) could only be detected up to 9 inch
for the revolver pistol. But, at the bullet ring, the
pink colouration could be observed by thoroughly
examination. This showed that the bullet contained
lead bearing material.
Scanning Electron Microscopy
(B)
SEM micrograph is made by examination of GSR
particle discharged from revolver pistol using
ammunition of Winchester .38 Special (Fig. 4).
(C)
Fig. 4: SEM micrograph for GSR particles discharged
from revolver pistol using Winchester 0.38 Special
ammunition at 7000x magnification.
(D)
Fig. 3: Pattern of GSR on cotton cloth fired using
revolver pistol at different firing distances (A) 3 inch, (B)
6 inch, (C) 9 inch and (D) 12 inch
It found that the particles appeared as irregular
cluster group grain of 3.3 µm in diameter. The
difference in geometrical shape of GSR particle is
possibly due to the different manufacturing process
that produced the kind of ammunition. Another
factor that could contribute to the different shape is
the fact that different process was involved in
formation of the particles for the semiautomatic
and revolver pistols. The particles undergo
different paths that gases and forming particulate
take upon escape from the pistols. In revolver pistol,
most of the gases from the primer burn escape in
the area around the cylinder, whereas in the
52
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
semiautomatic pistol, the initial path is the opening
provided by the ejection port [8].
Conclusion
Information gained from this study can be used to
estimate the muzzle-to-target distance for
reconstruction of shooting test up to 12 inch.
Chemical analyses involving modified Griess test
and sodium rhodizonate were successfully
employed. Results of modified Griess test
corroborated the visual examination as orange
specks revealing the presence of nitrite residues.
Whereas results indicated that both pistol discharge
contained lead from the positive colour change on
the cloth target. SEM micrograph from this study
revealed the geometrical shapes of the GSR
particles generated from different pistol and
ammunition.
Acknowledgements
Thanks are due to the Faculty of Science,
Universiti Teknologi Malaysia for laboratory
facilities, Royal Malaysian Police Forensic
Laboratory for assistance in the shooting tests and
Universiti Teknologi Mara Malaysia for financial
aid to Mohamed Izzharif Abd Halim for his
master’s study – Master of Science (Forensic
Science).
References
1.
2.
3.
Romolo, F. S. and Margot, P. (2001).
Identification of Gunshot Residue: A Critical
Review. Forensic Science International.
119:195-211.
Plattner, T. Kneubuehl, B. Thali, M. and
Zollinger, U. (2003). Gunshot Residue Patterns
on Skin Angled Contact and Near Contact
Gunshot
Wounds.
Forensic
Science
International. 138:68-74.
Brozek-Mucha, Z., Zadora, G. and Dane, F.
(2003). A Comparative Study of Gunshot
Residue Originating from 9 mm Luger
Ammunition from Various Procedures. Science
Justice Journal of the Forensic Science Society.
43: 229-235
4. Rosenberg, M. B. and Dockery, C. R. (2008).
Determining the Lifetime of Detectable
Amounts of Gunshot Residue on the Hands of
a Shooter Using Laser-Induced Breakdown
Spectroscopy. Applied Spectroscopy. 62:11
5. Bailey, J. A. Casanova, R. S. and Bufkin K.
(2006). A Method for Enhancing Gunshot
Residue Patterns on Dark and Multicoloured
Fabrics Compared with the Modified Griess
Test. Journal of Forensic Sciences. 51:4.
6. Haag, L. C. (2006). Shooting Incident
Reconstruction. San Diego, California:
Academic Press.
7. Muller, D. Levy, A. Vinokurov, A. Ravreby,
M. Shelef, R. Wolf, E. Eldar, B. Pharm, M.
and Glattstein, B. (2007). A Novel Method for
the Analysis of Discharged Smokeless Powder
Residues. Journal of Forensic Sciences. 52:1
8. Schwoeble, A. J. and Exline, D. L. (2000).
Current Methods in Forensic Gunshot Residue
Analysis. Boca Raton, Florida: CRC Press.
9. James, S. H. and Nordby, J. J. (2005).
Forensic Science – An Introduction and
Investigative Techniques. Boca Raton, Florida:
CRC Press
10. National Institute of Justice. Firearm Examiner
Training. The Research, Development, and
Evaluation Agency of the U.S. Department of
Justice.
Retrieved
from
www.ojp.usdoj.gov/nij/training/firearmstraining/module_12 on 10 January 2008.
11. D’Uffizi, M., Falso, G., Ingo, G. M. and
Padeletti, G. (2002). Microchemical and
micromorphological features of gunshot
residue observed by combined use of AFM,
SA-XPS and SEM + EDS. Surface and
Interface Analysis.
Additional information and reprint requests:
Mohamed Izzharif Bin Abdul Halim
(Email: izzharif@salam.uitm.edu.my)
Chemistry (Forensic Analysis) Program
Faculty of Applied Sciences
Universiti Teknologi MARA
40450 Shah Alam, Selangor, Malaysia
53
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
A Study on the Age Related Retention of Individual
Characteristics in Hand Writings and Signatures for Application
during Forensic Investigation
Nataraja Moorthy Ta, Mohamad Mahathir Amir Sultanb, Kong-Yong Wongc
a
Forensic Science Programme, School of Health Sciences,Universiti Sains Malaysia, Kelantan, Malaysia
b
Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia
c
Department of Chemistry Malaysia, Petaling Jaya, Malaysia
ABSTRACT: Handwriting is the means of communication and recording information in day-to-day life even
with the introduction of new technologies. Languages are systems of symbols; writing is a system for
symbolizing these symbols. Handwriting is quite distinctive [1]. Each individual’s writing style is personal and
unique which is the result of unconscious, automatic actions and interaction between a person’s brain, eye and
hand. Writing is also influenced by the person’s physical state [2], influence by drugs or alcohol [3], age [4], the
force on the writing tool [5] and other environmental factors [6]. An individual’s writing styles change
throughout the lifespan of a person and the degree of change may vary greatly. No two individuals, including
twins can write alike [7-8]. Investigators and forensic document examiners have long been tasked to use
inscriptions and markings as clues to identify their writers. This research aims to study the age related retention
of individual characteristics in handwritings and signatures over a period of time ranging from two to ten years.
Samples of seventy past handwritings and signatures were collected from individuals and compared with the
present handwritings and signatures of those individuals. Individual characteristics that were found to be
retained as well as those that were modified were identified and compared. The subjects were healthy and sound.
In the first step magnifiers were used to examine the writings and signatures and then comparison microscope
was used for detailed document examination. The analysis of writing style and signature characteristics
indicated changes attributable to deviation. In spite of these changes occurring due to age, the characteristics
that were retained are found to be useful in attributing authorship while examining documents.
Keywords: Forensic science, handwritings, signatures, individual characteristics, retention
Introduction
A document can be defined as an item that contain
writings, symbols, marks whether visible or
invisible and conveys a meaning for someone.
Document examination is defined as a discipline in
which a document is examined and analyzed in
order to obtain information that can be used to
serve the justice system based on the scientific
explanations [9]. Handwriting identification is a
process in which handwriting is used as a mean of
comparison and discrimination. In a more specific
manner, handwriting identification can be based on
evaluation of the significance of their similarities
and dissimilarities. The amount of significant
similarities and dissimilarities will be the cue in
concluding whether the handwriting belonged to a
particular person or not in cases like forgery and
fraud. Population studies have been conducted to
identify the class characteristics among the races
[10-12]. Signatures and handwriting have long
played a role in day-to-day activities like business
transactions, casual writing of letters, and listing of
required article in paper before going for shopping.
In forensic science they are used to authenticate
documents, as evidence to establish crime or
innocence, etc. This research aims to study the age
related retention of individual characteristics in
handwritings and signatures over a period of time
ranging from 2 to 10 years.
Materials and methods
(a) Sample collection
Samples of seventy past handwritings and
signatures from seventy volunteers were collected
and these samples were written or signed 2 to 10
years prior to collection. Sample handwritings and
signatures were correspondingly obtained from the
same volunteers. The age of the subjects ranged
from 20 to 35 years and they did not exhibit any
health problem that would affect handwritings. The
past samples designated as Q were serially
numbered as Q1 to Q70 while the present samples
designated as S were serially numbered as S1 to
S70. Regarding past samples, whenever more than
one signature or handwritings was collected from
the same individual, such writings were designated
54
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
with roman alphabet following the sample number
as chronological basis such as Q1a for the past
sample in an earlier year and Q1b for the past
sample in a later year both from the same
individuals.
The details of sample collections are as shown in
Table 1. The handwritings and signatures included
Bahasa Malaysia and English for both of which the
script is same.
Table 1: Details of sample collection
Sample No
Type of sample
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15 to 32
33 to 51,66
52 to 65
67 to 70
Handwriting
Handwriting
Signature
Handwriting
Handwriting
Handwriting
Handwriting
Handwriting
Handwriting
Handwriting
Handwriting
Handwriting
Handwriting
Handwriting
Handwriting
Signature
Handwriting
Handwriting
Year of past sample &
designated as Q
(presumed Questioned)
2000 (Q1a) , 2003 (Q1b)
2002 (Q2)
2002
2003 (Q4a) ,2005 (Q4b)
2003
2004
2004
2005
2005
2005
2005
2005
2004
2004
2006
2006
2006
2006
(b) Method of analysis
Magnifiers were used to compare the words and
signatures and then comparison microscope was
used for detailed examination. The next step is to
record the samples. This was done by mean of
scaled photography using a digital camera. The
writings were cropped using Adobe Photoshop,
enlarged 4 times its original size and were printed
using laser printer have been prepared for
comparison purposes. The original samples of
signatures and writings were examined very
carefully using hand magnifiers and the
characteristics were identified and recorded. The
Year of recent sample &
designated as S
(presumed Specimen)
2009 (S1)
2008 (S2)
2008
2008 (S4)
2009
2008
2008
2009
2008
2008
2009
2008
2008
2008
2009
2009
2009
2009
characteristics were marked in enlarged prints of
the writings obtained as stated before. Thus, the
original samples were preserved as such to prevent
damages and for future reference.
In the initial stage of examinations, words that
revealed multiple characteristics were chosen for
analysis. The identified characteristics were marked
and photographed both from past and recent
samples that were examined along side. Some of
the finding after analysis i.e. attribution of
authorship is as shown in Table 2 and in Fig. 1 to
Fig. 5.
Table 2: Findings after comparing some of the handwritings and signatures
Samp
No.
Fig
No.
Type of sample
2
1
Handwriting
3
2
Signature
4
3
Handwriting
6
4
Handwriting
66
5
Signature
Type of retained individual
characteristics
Findings after analysis
Design of letter, initiation, connection
between letters, termination
design, connection, stroke and manner
of dots
Letter design, connection between
letters, termination
Design of letters, initiation, connection
between letters, termination
Strokes, connection between letters,
termination, idiosyncrasy (smiley)
Both the handwriting belong to the
same person
Both the handwriting belong to the
same person
Both the handwriting belong to the
same person
Both the handwriting belong to the
same person
Both the signatures belong to the same
person
55
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Past year sample 2002 (Q2)
Recent year sample 2008 (S2)
Fig. 1: Comparison of sample No.2 [Handwriting]
Fig. 1 is a handwritten Bahasa Melayu sample. The
writings have been made in different years with a
span of six years i.e. 2002 (presumed questioned)
2008 (presumed specimen). The basic and overall
design of both the questioned and specimen
samples showed similarity and these observed
retention characteristics are indicated by arrows.
The letters “P”, “r”, “u” and “a” present in the past
and present samples showed retained individual
characteristics.
Past year sample 2002 (Q3)
Recent year sample 2008 (S3)
Fig. 2: Comparison of sample No.3 [Signature]
56
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Past year sub-sample (Q4)
2003 (Q4a)
2005 (Q4b)
Recent year sample (S4)
Fig. 3: Comparison of sample No.4 [Handwriting]
Past year sample 2004 (Q6)
Recent year sample 2008 (S6)
Fig. 4: Comparison of Sample No.6 [Handwriting]
57
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Past year sample 2006 (Q66)
Recent year sample 2009 (S66)
Fig. 5: Comparison of sample No.66 [Signature]
Results
All the present handwritings and signatures from
S1 to S70 were found to retain sufficient number of
individual characteristics, enabling comparison and
attribution of authorship with the corresponding
past handwritings and signatures. Some of the
observed finding results were shown in the table 2.
The table shows the sample number, the type of the
sample (handwriting or signature), the retained
individual characteristics which are identified (e.g.
design, stroke and termination).
Discussion
The handwritings and signatures are valuable
evidence in white collar offences during forensic
document investigation. In the samples of signature,
the retention of individual characteristics found to
be more pronounced in the stroke, design, location
of dots and connection. The characteristic retention
in signature is an important factor because
signature is one of the biometrics used for
verification or identification. There is also a sample
which show idiosyncrasy (sample 66) and it is
retained which increased the probability of positive
matching.
The minimum number of retained characteristics in
one word for past handwritings after a lapse of ten
years was 4 and maximum for the same ten years
period was 9. Similarly the minimum number of
retained characteristics in one word from
handwritings after a lapse of two years was 9 and
maximum for the same two years was 14
depending on the size of the samples. In general it
is found that the lesser the number of years for the
past writings, the more the number of retained
characteristics. However even after ten years, the
minimum number of retained characteristics for a
word and signature observed here were found to be
sufficient for attributing authorship indicating that
the individual characteristics are sufficiently
retained as a person become older. In this research,
it is found that all seventy samples of signatures
and writings showed retained individual
characteristics in the recent year samples.
Conclusion
This research was conducted on seventy samples of
handwriting and signatures to study the age related
retention of individual characteristics and its
forensic significance. The analysis was done on the
individual characteristics that were normally
assessed in forensics’ handwriting analysis. From
the study, it was found that even after ten years
most of the individual characteristics are retained.
Even though some changes were seen, still positive
identification could be made using the retained
individual characteristics. This research indicated
that individual characteristics such as initiation,
stroke, design of letters, location of dots,
connection between letters and termination are
more frequently retained even as time progresses.
This study indicated individual characteristics are
found to be retained in handwriting and signature
examination in spite of aging process.
Acknowledgements
The author is thankful to the volunteers for
providing the samples to conduct this research.
Acknowledgement is due to Dr. P. T. Jayaprakash,
Associate Professor, Forensic Science Programme,
Universiti Sains Malaysia, Kubang Kerian for his
support.
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
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changes in multi-finger interactions in adults
during maximkum voluntary finger force
production tasks. Human Movement Science.
27: 714-727.
Horton, R. A. (1996). A study of the
occurrence
of
certain
handwriting
characteristics in a random population.
International Journal of Forensic Document
Examiners. 2:95-102.
Franck, F. E. (2000). Handwriting is Unique:
Twin Studies. Presented at the American
Society of Questioned Document Examiners,
Ottawa, Canada.
8. Gamble, D. J. (1980). The handwriting of
identical twins. Journal of the Canadian
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9. Morris, R. (2000). Forensic handwriting
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Additional information and reprint requests:
Nataraja Moorthy T
(Email: nataraja@kb.usm.my)
Forensic Science Programme
School of Health Science
Universiti Sains Malaysia
16150 Kubang kerian, Kelantan, Malaysia
59
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Identifying Profile of Female Prisoners in Malaysian Prison
Using Carlson Psychological Survey
Nurul Hazrina Mazlana, Geshina Ayu Mat Saata, Affizal Ahmada
a
Forensic Science Programme, School of Health Sciences,Universiti Sains Malaysia, Kelantan, Malaysia
ABSTRACT: In Malaysia, the number of female prisoners seems to be increasing annually. To date, there has
been no empirical evidence regarding antisocial behavior, chemical abuse, or thought disturbance among female
prisoners in this country. With a goal to build a profile on female offenders in a Malaysian prison, empirical
knowledge on antisocial behavior, chemical abuse, thought disturbance, and self-deprecation are gathered in
through the Carlson Psychological Survey (CPS). There are two groups of females in this study; the study group
(n=134) consists of female prisoners from a female prison, and the control group (n=100) consists of females
from the society. Semi-structured survey was used in this study. Profile types were given to each respondent and
comparison was made between the study and control group. The findings concluded that female offenders have
a higher risk (α <0.05) of chemical abuse, thought disturbance, antisocial tendency, and self-depreciation than
females from the general population.
Keywords: Carlson Psychological Survey, female prisoners, profile.
Introduction
Female prisoners have always been a minor subject
in research and study. For a very long time, theory
and research relating to crime had been focusing on
male offenders [1, 2]. As a result, correctional
program that has been used for female offenders is
based on male criminality and often failed to
provide the appropriate option for the needs of
female offenders [1, 3]. The lack of information
and knowledge about them has caused difficulties
in designing an appropriate rehabilitation program
for this population. Although there is an existing
program for the rehabilitation of prisoners, a better
program may be created based on a better
understanding of the prisoners themselves. One
way to gather the information regarding the factors
for the appropriate rehabilitation is by creating a
profile of prisoners. This profile should consist of
empirical knowledge including antisocial behavior,
chemical abuse, thought disturbance, as well as
self-depreciation. This empirical knowledge can
give us an idea of why a woman commits a crime.
Furthermore, the existence and prevalence of
antisocial behavior among the female offenders can
be explored through the profile. Appropriate
profiling may result in the suggestion of a better
rehabilitation program for the female offenders.
The purpose of rehabilitation is to restore people
who offend to their previous or better condition.
This rehabilitation program can be implemented in
prison which is created to punish as well as to
rehabilitate a person who has offended. How well
this works depends on how effective the
rehabilitation program is. In order to create an
effective
rehabilitation
program,
sufficient
knowledge about the prisoners themselves is
needed. It is impossible to reach the complete and
effective rehabilitation for the prisoner without an
initiative and concentrated effort towards
appropriate diagnosis.
The objectives of this study are:
1. To build a profile of female prisoners in a
Malaysian prison.
2. To prepare empirical knowledge regarding
antisocial behavior, chemical abuse, thought
disturbance and self-depreciation.
3. To identify factors that may increase the
possibility of a female to commit an offence.
Method
Participant
This study involves two groups of respondent. The
first group is the study group (divided into 2
subgroups; Malaysian and non-Malaysian)
consisting of female prisoners from one prison
(named as prison A). For the Malaysian subgroup,
the numbers of respondents are 46 inmates, while
for the non-Malaysian subgroup, 88 respondents
participate in the study. The second group (n=100)
is the control group, the free living females in
Kubang Kerian and Kota Bharu, Kelantan.
Measure
Carlson Psychological Survey (CPS) designed by
Carlson (1982) [4], was used in this study. CPS is a
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
psychometric instrument designed specifically for
individuals who offend or are convicted of a crime.
It enables information regarding antisocial behavior,
chemical abuse and thought disturbance to be
gathered in one survey at one time. It has four
reliable scales which are chemical abuse (α=0.73),
thought disturbance (α=0.80), antisocial tendencies
(α=0.82) and self-depreciation (α=0.67). A validity
scale was included for the purpose of validating the
respondent test-taking attitude. This measurement
contains 50 items with a level four literacy level
which is easily understood by respondents.
Each item in CPS has five responses. The
respondent needs to choose one that is most related
to her and complete the 50 items. From each
response given, a raw score is calculated for each
scale and from each score, a profile is chosen.
These scores are transferred to a profile sheet to see
the profile pattern. The profiles are named as Type
1 to Type 18.
comparison between convictions in study group.
For statistical analysis, multivariate analysis of
variance (MANOVA) was used. For the profile
analysis, the data from the SPSS were transferred
to a scoring sheet. From the scoring sheet, the raw
score for each scale (chemical abuse, antisocial
tendencies, thought disturbance, self-depreciation
and validity) was calculated. The raw score was
then referred to a reference guide for classification
and the profile type was obtained. From the raw
score also, a profile sheet was filled and the pattern
of score observed. The main objective in using
MANOVA is to determine the affect of
independent variables on the dependent variables.
In this study, the independent variables were
ethnicity, age, type of crime and length of
incarceration for the study group. For the control
group, the independent variables were ethnicity,
age and working status. For dependent variables,
scores of chemical abuse (CA), thought disturbance
(TD), antisocial tendency (AT) and selfdepreciation (SD) were selected for both groups.
Data collection
The CPS was prepared according to the number of
participants. The study group answered the survey
in groups of ten in prison A. The instrument was in
English version. Survey was semi-structured and
the researcher had a chance of interacting with the
respondents but not directly interviewing the
respondents. In some cases where the respondents
were unable to understand the language, Malay was
considered. For the control group, participants who
volunteered to participate were given the survey,
and asked to return it as soon as they finished the
survey.
Analysis
The analyses included a comparison between the
study group and the control group, comparisons
between subgroups within the study group, and
Result and Discussion
Demographic information
According to Table 1, Indonesian had the highest
percentage (68%) followed by Malay (19%).
According to age, most of Malaysian (12%) was
within the age range of 20 to 29 and the same was
for the non-Malaysian subgroup (28%). For the
length of incarceration, most were sentenced within
six months to 12 months length (47%).
As shown in Table 2, most of respondents within
the control group were Malays (76%). For range of
age, most of the respondents were within 20 to 29
years old (64%). For working status, most of the
respondents were students (48%) followed closely
by working in various occupations (45%).
Table 1: Summary of demographic information for the study group which includes ethnicity, age and length of incarceration
Ethnicity
Age
Length of
incarceration
Demographic Information
Malay
Chinese
Indian
Indonesian
20 to 29
Malaysian subgroup
Non Malaysian subgroup
30 to 39
Malaysian
Non Malaysian
40 to 49
Malaysian
Non Malaysian
50 and above Malaysian
Non Malaysian
Less than 6 months
6 months to 12 months
More than 12 months
Frequency
27
10
8
97
17
40
11
35
15
19
3
2
45
65
32
Percentage
19.00
7.00
5.60
68.30
11.97
28.17
7.75
24.65
10.56
13.38
2.11
1.41
31.00
46.50
22.50
Total
142
57
46
34
5
142
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Table 2: Summary of demographic information for the control group which includes ethnicity, age and working status
Demographic Information
Malay
Ethnicity
Chinese
Indian
20 to 29
30 to 39
Age
40 to 49
50 and above
Working
Working status
Non Working
Student
Profile Type
According to Table 3, 13 profile types were
categorized in this study. Some of the results were
unidentified as there were no suitable profile types
for the raw score as per the CPS manual. The
suitable profile for these scores has not been
produced. Comparison was made between means
of the usable scores from this study to the means
given by the CPS manual.
Profile type 5 was the most frequent profile type
for the Malaysian incarcerated subgroup (17%).
This means, these individuals have high antisocial
tendency and poor social skills. Attention should be
given to these individuals since suicidal thoughts
are possible amongst them. Profile type 6 and 7
were the second most frequent (15 %) amongst the
study subgroup. Profile type 6 is indicated by a
passive-aggressive personality. People with this
profile type show no remorse for their offences and
often denying responsibility. Profile type 7
demonstrates a disturbed personality and
psychiatric treatment is recommended for this
group. The other profile types are less frequent as
compared to these three profile types.
Among the non-Malaysian subgroup, most of them
(62.5 %) have unidentified profile type. As
mentioned earlier, unidentified means that the
profile types suitable for the scores of individuals
in this group has not been produced. This is likely
as these individuals did not have any drug or
antisocial related offences. Profile type 7 and 11
were the second highest profile types (16%).
Profile type 11 is generally emotionally passive and
is likely to be influenced by others. People of this
profile may engage in depression since they
demonstrate restlessness and strange behaviors. A
high score in antisocial tendency shows that this
group is at risk of having antisocial behavior. This
Frequency
76
11
13
64
16
16
4
45
7
48
Percentage
76
11
13
64
16
16
4
45
7
48
Total
100
100
100
profile type was the second highest for the control
group as well (35%).
Most of the respondents (44%) in the control group
have unidentified profile type. This was expected
as the control group represented the normal
population. It was assumed that people from the
normal population are generally less likely to have
antisocial behavior or indulge in activities related
to chemical abuse.
An overview on the result of profile type showed
that there are differences among groups. The
Malaysian subgroup has a set of profile type which
is different from the non-Malaysian subgroup and
the control group. This indicates that female
offenders in the Malaysia subgroup have different
characteristics and this can be explained through
their profile types. This also may be useful to show
differences between the offender population and
the non-offending population.
Statistical analysis
According to Table 4, it showed that chemical
abuse and antisocial tendency are very common
among the offenders population. Both factors
occurred across the ethnicity, the types of crimes
and the length of incarceration. Chemical abuse
occurred across the age range as well. This proved
that chemical abuse and antisocial behavior are
high among offenders [6, 7, 8]. The occurrence of
chemical abuse and antisocial among this
population also showed that there is a positive
association between these two factors [9, 10, 11].
Through this study, thought disturbance and selfdepreciation did not show a strong occurrence
amongst the offender population. However, this
does not indicate that there is no thought
disturbance and self-depreciation in the entire
population.
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Table 3: Summary of the profile type for both groups
Groups
Study group
Malaysian
subgroup
Non
Malaysian
Subgroup
Control group
Profile Type
1
2
3
5
6
7
8
9
10
17
18
Unidentified
6
7
10
11
13
Unidentified
6
7
11
13
Unidentified
The control group did not show any strong
prevalence of all these factors except across the
ethnicity. Between ethnicities, there are occurrence
of chemical abuse and self-depreciation. However,
comparison of means for chemical abuse between
the study group and the control group showed that
the control group has much lower mean than the
study group. This indicates that there are
occurrences of chemical abuse in the control group.
However, incidences might be low and not
significant for this study.
Comparison between groups, including the
subgroups showed a positive result. As expected
for the outcome, there is a prevalence of all the
dependent variables amongst the study group. The
Malaysian subgroup showed a high prevalence of
chemical abuse and antisocial tendency. On the
other hand, non-Malaysian subgroup showed a high
prevalence of thought disturbance and selfdepreciation. This showed that the offender
population has higher a risk of chemical abuse,
antisocial tendency, thought disturbance and selfdepreciation compared to the general population [6,
7, 8].
Abuse of drug is significantly related to crime
against property and the involvement in criminal
activities is likely to support their drugs
dependency. Criminal behavior among drug abuser
Frequency
2
2
2
8
7
7
1
1
5
3
3
5
2
15
2
15
2
60
7
3
35
11
44
Percentage
4.35
4.35
4.35
17.40
15.22
15.22
2.17
2.17
10.87
6.50
6.50
10.87
2.08
15.63
2.08
15.63
2.08
62.50
7.00
3.00
35.00
11.00
44.00
Total
46
96
100
is most likely as a result of influence by friends as
well as poverty [12]. Therefore, aside from illegal
use of drug, they might have been involved in
property crime as well.
As mentioned earlier, there are positive association
of chemical abuse and antisocial tendency amongst
the Malaysian subgroup. Antisocial behavior had
been defined as related to substance dependence
and legal problems [13]. The result of this study
showed that there is a relationship between
antisocial behavior and chemical abuse. The high
prevalence amongst offenders showed that
antisocial behavior is likely to be related to
unlawful behavior as well.
Antisocial behavior had been described to include
violations of rules and social norms, various forms
of aggression, serious patterns of disruptive and
aggressive behavior and lack of guilt [14, 15].
People with antisocial behavior are more likely to
be involved in a more serious and violent crime due
to the aggressive nature and the lack of guilt. To
illustrate, drug abuser is more likely to involve in
property crime. There is possibility for more
violent crime to happen during the commission of
said crime if the drug abuser also has antisocial
attitude. When there is obstruction during robbery,
another crime may occur such as murder.
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
Table 4: Summary of the MANOVA tests on all independent and dependent variables in this study for both groups
Independent and
dependent variables
interaction
Ethnicity – all the
dependent variables
MANOVA Tests
1. Multivariate tests
 α = .000, significance differences in the dependent variables across ethnicity
2. Tests of BetweenSubjects Effects
 α for CA and AT < .05, CA and AT differ significantly across ethnicity.
 Due to insufficient numbers of sample for each ethnic, comparison among
ethnics cannot be done.
3. Estimated Marginal
Means
1. Multivariate tests
 CA and AT are likely to occur among ethnic in this group.
2. Tests of BetweenSubjects Effects
 α for CA < .05, CA differs significantly across age range.
 Due to insufficient numbers of sample, comparison among age range cannot
be done.
3. Estimated Marginal
Means
1. Multivariate tests
 CA is likely to occur among age range.
2. Tests of BetweenSubjects Effects
 α for CA and AT < .05, CA and AT differ significantly across the types of
crime.
 Due to insufficient numbers of sample for each types of crime, comparison
among them cannot be done.
3. Estimated Marginal
Means
1. Multivariate tests
 CA and AT are likely to occur among types of crime.
2. Tests of BetweenSubjects Effects
 α for CA and AT < .05, CA and AT differ significantly across the length of
incarceration.
 Due to insufficient numbers of sample, comparison among length of
incarceration cannot be done.
3. Estimated Marginal
Means
1. Multivariate tests
 CA and AT are likely to occur among different length of incarceration in
this group.
 α = .00, significance differences in the dependent variables across the
ethnicity
2. Tests of BetweenSubjects Effects
 α for CA and SD < .05, CA and SD differ significantly across ethnicity.
 Due to insufficient numbers of sample for each ethnic, comparison among
ethnics cannot be done.
3. Estimated Marginal
Means
1. Multivariate tests
 CA and SD are likely to occur among ethnic in this group.
2. Tests of BetweenSubjects Effects
 all the α values are more than .05, no dependant variables contribute to the
significant multivariate effects. Dependent variables are not affected by the
age range.
 all α values are more than .05, no significant multivariate effects in working
status.
Study group
Age range – all the
dependent variables
Types of crime – all
the dependent
variables
Length of
incarceration – all
the dependent
variables
Control group
Ethnicity – all the
dependent variables
Age range – all the
dependent variables
1. Multivariate tests
Between groups
Working status – all
the dependent
variables
Groups – all the
dependent variables
Results
 α >.05, no significance differences in the dependent variables across age
range
 α = .000, significance differences in the dependent variables across the
types of crime
 α = .000, significance differences in the dependent variables across the
length of incarceration.
 all α values are more than .05, no significant multivariate effects in age
range.
1. Multivariate tests
 all the α values are more than .05, no dependant variables contribute to the
significant multivariate effects. Dependent variables are not affected by the
working status.
 α = .000, significance differences in the dependent variables across groups.
2. Tests of BetweenSubjects Effects
 α for all dependent variables < .05, all dependent variables differ
significantly across groups.
3. Estimated Marginal
Means
 Malaysian subgroup has the highest means for CA and AT (means= 23.96,
32.98). Malaysian subgroup has higher score in CA and AT than the others.
 Non-Malaysian subgroup has highest means for TD and SD. The nonMalaysian subgroup score higher in both compare to the others.
2. Tests of BetweenSubjects Effects
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Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
The non-Malaysian subgroup showed a high
occurrence of thought disturbance and selfdepreciation compared to the Malaysian subgroup.
This may indicate that as a group non-Malaysian
incarcerated offender have some forms of mental
problems. Thought disturbance has been defined as
disturbance in the form of thought and may involve
confusion, hallucination, and delusion [16]. The
type of crime that might be committed by people
with thought disturbance is less likely to be a
violent crime or property crime. The high
prevalence amongst the non-Malaysian subgroup
may refer more to disturbance in thought due to
pressure being away from home, which is
Indonesia. Self-depreciation by its nature does not
seem to predispose an individual to antisocial
related crimes. Self-depreciation is defined as
undervalue of oneself and it is more likely due to
the feeling of inferiority and weakness. People with
high self-depreciation are more likely to be a
victim rather than a criminal.
High prevalence of thought disturbance and selfdepreciation amongst the non-Malaysian subgroup
does not mean the Malaysian subgroup does not
suffer from these two factors. In fact, the means of
both variables for the Malaysian subgroup are very
close to the non-Malaysian. They are also very high
compared to the control group. This showed that
there are likelihoods that the Malaysian subgroup
may have thought disturbance and self-depreciation
as well.
Conclusion
The profiles of female offenders have been
successfully generated at the end of this study. It
shows a set of profiles which differs from the
control group. This profile types are unique to each
offender and this proves that every offender has
different needs for rehabilitation, as mentioned in
the introduction. Each profile type contains all the
empirical knowledge needed and explains what is
necessary for a better rehabilitation.
The analysis carried out evidenced the remaining
objectives in this study. Chemical abuse and
antisocial tendency have been proved to be high
amongst the female offenders. These showed that
both variables are very likely to become the factors
for a female to commit a crime. Both factors also
showed a high probability to be associated with
each other. A comparative analysis proved that
chemical abuse and antisocial tendency are more
likely to occur in the offender population compared
to the general population.
subgroup. However, this does not mean that both
do not occur amongst the Malaysian subgroup. The
result of the analysis showed that the Malaysian
subgroup has a high probability to suffer thought
disturbance and self-depreciation as compared to
the control group. Therefore, the offender
population is more likely to have thought
disturbance and self-depreciation. Both are also
likely to become the factors for a female to offend.
Limitations of the Study
The first limitation is the CPS. The CPS is prepared
based on studies done in America. The baseline
means and standard deviation are based on such
studies. As far as this study is concerned, this
survey is not fully applicable to the Malaysian
population. While it may produce some good
results, however not all the results were obtained as
anticipated. An example is, the profile type which
cannot be identified. This may be due to the
limitation in the scale created by Carlson which is
not fully suitable in Malaysia. Furthermore, items
in CPS may not be as relevant to the Malaysian
environment. There is a possibility that some
behavior unique to the Malaysian offending
population were excluded.
The second limitation is the interaction with the
respondents. This study was done based on a semistructure study. It involved limited interaction
between the researcher and the respondents. Due to
this, there is a probability that the respondents
could not answer the CPS to their best ability. The
third limitation is language, mainly for the nonMalaysian subgroup. Differences in some
vocabulary and use of words limit the proper
interaction between the researcher and the
respondents. There is possibility that the
respondents in this group were unable to fully
understand the questions given.
Acknowledgements
The authors thank Jabatan Penjara Malaysia for
giving the chance to conduct this study.
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Additional information and reprint requests:
Affizal Ahmad
(Email: affizal@kb.usm.my)
Forensic Science Programme
School of Health Sciences
Universiti Sains Malaysia
16150 Kubang Kerian, Kelantan, Malaysia
66
Malaysian Journal of Forensic Sciences, 2010, Vol. 1, No. 1
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References: References should be based on sequential numerals in square brackets [3]. All references should be
listed in numerical order in the Reference section with the format authors, years, title of paper, journal title,
volume, issue number, and pages.
Example: Andrasko J., Stahling S. (1999). Time since discharge of spent cartridges. Journal of Forensic Sciences 44(3):
487-95.
Copyright: Submission of a manuscript implies that the work has not been published before, not under
consideration for publication elsewhere, and has been approved by all-co-authors.
Proofs: PDF page proofs will be emailed to authors for checking, and should be returned within 5 days to the
Editor.
Typescripts: Malaysia Journal of Forensic Sciences operates an on-line submission and reviewing system.
Authors should log on in order to submit an article at www.forensics.org.my. If any difficulties are encountered,
please contact
Editor-in-Chief
Malaysia Journal of Forensic Sciences
Email: mjofseditor@gmail.com
Subscriptions
To subscribe Malaysia Journal of Forensic Sciences, please register here. Subscription (for June 2010 issue) is
free.
Malaysia Journal of Forensic Sciences is unclassified (as of October 2010), and is published electronically on
the Forensic Science Society of Malaysia at www.forensics.org.my and hence require Internet access to view.
The publication itself is never sent electronically (that is, as attachments). Access to Malaysia Journal of
Forensic Sciences is free (for October 2010 issue).
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