Forensic Science International 302 (2019) 109911

Contents lists available at ScienceDirect

Forensic Science International

journal homepage: www.elsevier.com/locate/forsciint

Review of the most common chemometric techniques in

illicit drug profiling
Ana Popovica , Marie Morelatoa , Claude Rouxa , Alison Beavisa,b,*
a
Centre for Forensic Science, University of Technology Sydney, P.O. Box 123 Broadway NSW 2007 Australia
b
Faculty of Transdisciplinary Innovation, University of Technology Sydney, P.O. Box 123 Broadway NSW 2007 Australia

A R T I C L E I N F O A B S T R A C T

Article history: The information generated through drug profiling can be used to infer a common source between one or
Received 22 January 2019 several seizures as well as drug trafficking routes to provide insights into drug markets. Although well
Received in revised form 19 July 2019 established, it is time-consuming and ineffective to compare all drug profiles manually. In recent years,
Accepted 29 July 2019
there has been a push to automate processes to enable a more efficient comparison of illicit drug
Available online 1 August 2019
specimens. Various chemometric methods have been employed to compare and interpret forensic case
data promptly. The intelligence that is produced can be used by decision-makers to disrupt or reduce the
Keywords:
impact of illicit drug markets. This review highlights the most common chemometric techniques used in
Pattern recognition
Comparison metrics
drug profiling and more specifically, the most efficient comparison metrics and pattern recognition
Forensic intelligence techniques outlined in the literature.
© 2019 Elsevier B.V. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2. Drug profiling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.1. Profiling of chemical characteristics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.1.1. ATS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.1.2. Cocaine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.1.3. Heroin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.2. Profiling of physical characteristics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3. Chemometric techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
3.1. Pre-treatments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
3.2. Comparison metrics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
3.2.1. Square cosine function . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
3.2.2. Pearson correlation coefficient . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
3.2.3. Euclidean distance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
3.2.4. Manhattan distance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
3.2.5. Canberra distance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
3.3. Unsupervised pattern recognition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
3.3.1. Principal component analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
3.3.2. Hierarchical clustering analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
3.3.3. K-means clustering . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
3.4. Supervised pattern recognition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.4.1. Partial least squares – discriminant analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.4.2. Linear discriminant analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.4.3. Artificial neural networks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
4. Evaluation of chemometric techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
4.1. Evaluation of CMs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

* Corresponding author at: Centre for Forensic Science, University of Technology, P.O. Box 123 Broadway, Sydney, NSW 2007, Australia.
E-mail address: Alison.Beavis@uts.edu.au (A. Beavis).

http://dx.doi.org/10.1016/j.forsciint.2019.109911
0379-0738/© 2019 Elsevier B.V. All rights reserved.
2 A. Popovic et al. / Forensic Science International 302 (2019) 109911

4.1.1. Discrimination power . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

4.1.2. Confusion matrices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
4.1.3. Receiver operating characteristic curves . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
4.2. Evaluation of UPR techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
4.2.1. PCA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
4.2.2. Cluster validation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
4.3. Evaluation of SPR techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
5. Application and added value of chemometric techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.1. Applications of CMs to illicit drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.1.1. ATS (AM & MA) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.1.2. ATS (MDMA) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.1.3. Cocaine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
5.1.4. Heroin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
5.2. Applications of UPR to illicit drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
5.2.1. ATS (AM, MA & MDMA) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
5.2.2. Cocaine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5.2.3. Heroin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5.3. Applications of SPR to illicit drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
5.3.1. ATS (AM, MA & MDMA) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
5.3.2. Cocaine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
5.3.3. Heroin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
5.4. Added value of chemometric techniques applied to illicit drugs for ILP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
5.4.1. Added value for active investigations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
5.4.2. Added value for understanding drug markets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
6. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
CRediT authorship contribution statement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15

1. Introduction performed on illicit drugs to produce chemical and physical

Illicit drug markets are continually evolving, so it is essential to
understand them in order to disrupt or reduce their impact.
Decisions by law enforcement agencies regarding crime disrup-
tion, prevention and reduction rely on masses of information from
various sources, which is often minimally exploited. The concept of
forensic intelligence focuses on generating information products
for proactive policing, supporting an intelligence-led model rather
than focusing on individual offenders or cases.
A variety of analyses (qualitative and quantitative) are currently
performed on illicit drugs to generate profiles [1]. These profiles, in
combination with chemometric techniques, can be used proac-
tively and generate intelligence products at tactical, operational
and strategic levels [2,3].
Drug intelligence has multiple aims [4–7], for example they are
to (1) identify dealer–user networks; (2) determine the geograph-
ical origin of the seizure; (3) monitor the length of time the
clandestine laboratory has been in operation; (4) gather sufficient
information to create national and international drug databases;
(5) monitor the extent of national and international drug
trafficking; (6) obtain a better understanding of drug networks
and (7) ultimately develop sound strategies to reduce the harm
caused to the society.
To maximise the potential of drug profiling, extensive efforts
have been invested in harmonising analytical methods to
facilitate the comparison of data generated from different
laboratories [8–13]. Visual comparison of profiles is still routinely
used; however, the task becomes laborious and inefficient when
applied to a database of specimens [14,15]. Additionally, the
visual analysis does not allow for an overall view of the drug
problem; hence, it is of limited value for intelligence purposes.
For this reason, drug analysis laboratories have sought out to
automate the comparison processes via mathematical and
statistical methods.
This article aims to review the various chemometric methods
used in drug profiling. Section 2 will explore the current analyses
profiles. Section 3 will explore common chemometrics techniques
used in drug profiling and discuss how they work. Section 4
reviews possible evaluation tools for the chemometric techniques
outlined in Section 3. While Section 5 will look at the application of
these chemometric techniques to drug profiling data and their
added value for intelligence-led policing (ILP).
2. Drug profiling
A drug profile contains information about the chemical or/and
physical characteristics of a specimen. The chemical profile yields
data about the illicit substance along with cutting agents
(adulterants and diluents), precursors, by-products, impurities
and solvents in different ratios. The physical profile (packaging and
appearance) of a drug can be just as complex, creating a particular
picture of a specimen when all measurements are considered [16].
It is important to note that the selection of target variables for
the analytical method will profoundly influence the results
obtained from the chemometric methods applied [17]. There are
criteria that need to be satisfied when choosing target variables:
(1) they must be present in most specimens and have sufficient
concentration; (2) they must be sufficiently numerous to group
specimens with similar profiles and differentiate unrelated speci-
mens; (3) their measurement must be reproducible and repeat-
able; and (4) they should be representative of the specimen to be
described [8].
2.1. Profiling of chemical characteristics
The subsequent sections outline the most common analytical
techniques used to generate chemical profiles for amphetamine-
type stimulants (ATS), cocaine and heroin specimens.
2.1.1. ATS
As many ATS exist as optical isomers, and they differ in
biological activity, it is essential to determine the concentration of
 A. Popovic et al. / Forensic Science International 302 (2019) 109911 3

each enantiomer. This can be achieved using capillary electropho- Table 2

Common analytical methods used in cocaine profiling.
resis (CE) [18]. An elemental profile can be generated using
inductively coupled plasma mass spectrometry (ICP-MS). As most Profile type Analytical method References
of the synthetic pathways to illicit drugs employ catalysts, a Adulterants & diluents ATR-FTIR [52,53]
knowledge of the trace elements present in either 3,4-methyl- Alkaloids GC-MS [17,54,55]
enedioxy-methamphetamine (MDMA) or methylamphetamine Alkaloids GC-NPD [45]
Elements ICP-MS [56]
(MA) can potentially reveal the route of synthesis [19–21].
Isotope ratios IRMS [50,51]
Manufacturing by-products can be determined through a tech- Solvents HS GC-MS [47,49]
nique involving a liquid-liquid extraction (LLE) or headspace solid-
phase microextraction (HS-SPME) followed by gas chromatogra-
phy–mass spectrometry (GC–MS) or gas chromatography-flame
ionisation detection (GC-FID) analysis [22–25]. The presence of Table 3
impurities or by-products provides information on trafficking Common analytical methods used in heroin profiling.
routes, supply origin and optimally link seizures [20].
Profile type Analytical method References
Additionally, The Collaborative Harmonisation of Methods for
Alkaloids CE [63]
Profiling of Amphetamine-Type Stimulants (CHAMP) project
By-products GC-MS [13,17,62,63,66,67]
conducted a significant study on developing a harmonised By-products GC-FID [60,61,68,69,70,71,72,73]
profiling method for ATS [26–31]. Adulterants and diluents are Elements ICP-MS [74,75,76]
also frequently encountered in ATS specimens. Conventional Solvents HS GC-MS [49,63]
analysis techniques for these profile types are capillary electro-
phoresis diode array detector (CE-DAD) and liquid chromatogra-
phy evaporative light scattering detector (LC-ELSD) [32,33]. Stable
isotope ratio – mass spectrometry (IRMS) analysis also forms an which more than a hundred have been identified and characterised
integral part of ATS profiling. The 13C/12C, 15N/14N and D/H ratios of [62]. This type of profile once again aims to determine the origin of
a specimen can infer the route of synthesis and subsequently links the plant [62–65] and is an adjunct of the significant alkaloid
to other specimens [18]. The mentioned ATS profile types and their profiling. As with cocaine, during the manufacturing of heroin, a
relevant references have been summarised in Table 1. range of solvents are employed which are determined using HS
GC–MS [49]. This information can provide insights into where
2.1.2. Cocaine processing occurred [46]. This is because solvent combinations
Determining the presence of coca leaf alkaloids is typically the often differ between major drug production regions.
first step when profiling cocaine, see Table 2 for a list of common
cocaine profile types and relevant references. This information is 2.2. Profiling of physical characteristics
useful for determining the origin of the plant. There have been
several articles that have quantified cocaine alkaloids using GC–MS Physical profiling looks at the appearance and packaging of
or a gas chromatography – nitrogen phosphorous detector (GC- illicit drugs and provides complementary information to respec-
NPD) [42–45]. Another profile for cocaine involves analysing the tive chemical profiles. In terms of collection, physical profiles are
residual solvents present, indicating the process used to convert timelier and more resource friendly than their chemical counter-
the drug base to the salt form [46]. Solvent traces can be easily parts. Furthermore, in an instance where chemical and physical
detected using GC–MS or headspace (HS) GC-FID [47–49]. (including packaging) profiles were evaluated, the latter proved
Further information regarding location can be determined by to be pivotal for supporting ILP [77]. This is especially true for
the ratio of carbon and nitrogen stable isotopes [50,51]. For optimal specimens in a pill form. In the literature, the analysis and
linkage capacity, the mentioned method should be used with a evaluation of physical profiles, appearance in particular, is mainly
significant component (alkaloid) analysis. As cocaine is often cut applied to MDMA tablets. A visual inspection of a specimen’s
with various compounds, adulterant and diluent analysis can be physical characteristics can allow seizures to be subcategorised
performed via vibrational spectroscopy, namely attenuated total based on colour, texture, score/logo presence, measurements, or
reflection Fourier transform infrared (ATR-FTIR) spectroscopy general appearance [11,72,78]. The score/logo and tablet measure-
[52,53]. ments are machine dependent and tend to remain unaltered
between batches, allowing for the potential to link seizures [79].
2.1.3. Heroin Furthermore, a closer examination of packaging material may infer
As with cocaine, the initial analyses performed aim to identify links between chemically unrelated seizures. Table 4 indicates the
the alkaloids present, see Table 3 for a list of common heroin profile current articles focusing on profiling of physical characteristics for
types and relevant references. Remaining analytes, if present, are drug intelligence purposes.
normalised to the heroin response. These types of methods The visual inspection of a specimen is usually performed before
are relatively fast, but they lack sensitivity [57–61]. Throughout the any chemical analyses. This is due to the ease and accessibility
heroin production process, many by-products are generated, of of visual characteristics. For MDMA, physical characteristics (i.e.

Table 1 Table 4
Common analytical methods used in ATS profiling. Current articles focusing on physical profiling for drug intelligence purposes.

Profile type Analytical method References Sample type Analytical method References
By-products GC-MS [10,12,25,31,32,33,34,35,36,37,38,39] MDMA Visual [11,79,80]
By-products GC-FID [40,41] Heroin Visual [72]
Adulterants CE-DAD [32,33] Adhesive tape Py-GC-MS [81]
Diluents LC-ELSD [32,33] Adhesive tape VSC [82]
Elements ICP-MS [32,33] Adhesive tape SFE-HPLC [83]
Enantiomers CE [18] Adhesive tape MSP [84]
Isotope ratios IRMS [18] Heroin Packaging General measurements [72]
4 A. Popovic et al. / Forensic Science International 302 (2019) 109911
post-tabletting characteristics) may yield preliminary profiling
data while chemical characteristics (i.e. pre-tabletting character-
istics) provide complementary data. Together they allow speci-
mens to be linked at multiple stages of production [80].
As the two sets of characteristics (physical and chemical) are
attributed at different stages of the MDMA production process,
the individual profiles are complementary [11]. Any similarity in
physical characteristics only goes so far as to suggest MDMA
specimens were tabletted in the same laboratory; common origin
at a pre-tabletting stage should not be inferred [78]. On the other
hand, specimens with differences in physical characteristics may
still be linked at a chemical level. Manufacturers of drug powders
may tablet the same batch at different times, altering moulds in
between or the tabletting step may be conducted in a different
location [78].
Chan et al. [72] hypothesised that, for heroin, the distributor's
trademark was the colour of the specimen. The addition of
adulterants and food dyes added during cutting would allow the
distributor and an easy way to differentiate between different
batches and manufacturers.
Although the use of packaging material in a drug intelligence
perspective may be constrained, it can still provide some insight
into how a criminal organisation prepares its goods for transport
[72,81,85]. The primary constraint is the fact that the packaging
profile may be short-lived, as the materials are readily available.
The benefit of packaging profiling is that it can be used to link
seizures where the drug profile is not useful [81]. Significant drug
seizures often contain subgroups with different impurity profiles,
either due to batch intra-variability or sporadic manufacturing
[78]. Subsequently, linking objects based on chemical character-
istics may prove challenging. In this instance, linking seizures may
only be possible with physical characteristics (appearance and
packaging). The packaging profile can include the dimensions,
colour or general appearance of the package. Furthermore, any
markings on the package, such as stamps, can infer the geographic
location of the sender [85]. Rhumorbarbe et al. [85] also evaluated
the use of physical profiling for determining the accuracy of online
purchase information. It was found that the digital and physical
profiling information were complementary and aided the evalua-
tion of cryptomarket activity.
Additionally, research conducted in the field of physical
profiling also includes the development of analytical techniques
for profiling common packaging materials [78,81–83]. In a three-
part study, Huttunen et al. investigated whether pyrolysis GC–MS
(Py-GC–MS) [81], a Video Spectral Comparator (VSC) [82] and
supercritical fluid extraction with high-performance liquid chro-
matography (SFE-HPLC) [83] could be used to differentiate
adhesive tapes. Although these analytical techniques are chemical,
the resultant profiles ultimately describe a physical entity of the
seized specimen.
Some limitations need to be taken into account when analysing
and evaluating drug packaging [81]. It is entirely possible that
two independent laboratories might be using the same packing
material, hence introducing FPs. Furthermore, changes noticed in
the chemical composition of packaging may be due to a change in
production by the manufacturer. For this reason, the focus of
seizure comparison made through physical profiling should be
relational. That is, rather than aiming to identify the manufacturer
of the packaging and thereby inferring links, analysts should aim to
compare physical profiles to existing physical profiles from
previous seizures.
3. Chemometric techniques
Forensic scientists utilise a multitude of instruments for the
analysis of traces which generate large and complex datasets. These
datasets are subjected to various mathematical and statistical
techniques, which include comparison metrics (CM) for determining
links between objects, pattern recognition for the classification of
objects and the class prediction of new, unclassified objects.
In the case of illicit drug profiles, it is common to ‘pre-treat’ the
data resultant from qualitative or quantitative analyses [76].
Subsequently, metrics which calculate the similarity between
objects are applied to drug data to determine the level of
association between them.
3.1. Pre-treatments
Analyses obtained from two different instruments may not
exhibit the same sensitivity. Pre-treatments (PT) are employed to
reduce the instrumental influence, and therefore represent an
essential part of the overall profiling process [31,86,87]. Also, PTs
are used to minimise the effect of significant differences in the
concentration of compounds present in a specimen [76]. The most
common PTs are outlined inTable 5. This is not an exhaustive list of all
possible PTs, for a more thorough list, please refer to Lociciro et al. [9].
In these various PT approaches, xi is the untreated peak area of
compound i; Ni and Si are the normalised and standardised peak
areas of compound i, respectively.
PTs are typically used in combination with CMs, which are
discussed below. Various PTs have been documented. For instance,
Perkal [88] normalised GC data before comparison of MA
specimens; Klemenc [66] employed normalisation and weighting
when comparing different heroin seizures; Jonson [89] weighted
and normalised data for comparison of amphetamine specimens.
3.2. Comparison metrics
To determine the level of association between profiles, CMs
need to be used. The metrics that will be outlined in this article are
the Square Cosine Function (including modifications), Pearson
Correlation (including modifications), Euclidean distance, Man-
hattan distance and the Canberra distance. There are alternative
techniques, but the five CMs explored in this article (see
Sections 3.2.1–3.2.5) are utilised most frequently [31,69].
These CMs produce a score which represents the similarity/
distance between objects or in this context, illicit drug profiles.
After the drug profiles are separated into two populations, i.e.
linked and unlinked, scores are calculated for each pair of profiles.
The ideal statistical combination of PT and CM must be robust in
differentiating between linked and unlinked populations.
To create the linked population, all specimens in a seizure are
compared against each other. This process is repeated for every
seizure containing more than one specimen. Doing this allows for
the assessment of the intra-variability of a seizure, establishing the
level of similarity between specimens of an assumed common
origin. Similarly, to create the unlinked population, specimens
from different seizures are compared to each other. This
determines the inter-variability between seizures or the level of
similarity between specimens of an assumed different origin [8].
Table 5
Most common PT methods.
PT (abbreviation) Formula
Normalisation (N) Ni ¼ Pxn i
xi
Standardisation (S) xi
i¼1
Si ¼ S:D:i
Logarithm (L) logðxi Þ
p ffiffiffiffi
Fourth root (4R) 4
xi
Normalisation + standardisation (N + S) Ni
S:D:i
Normalisation + fourth root (N + 4R) p
4
ffiffiffiffiffi
Ni
 A. Popovic et al. / Forensic Science International 302 (2019) 109911 5

score of 100 means that the specimen pair has a low correlation.
The formula for the modified SCF (MCF) is:
MCF = 100  SCF

3.2.2. Pearson correlation coefficient

The Pearson correlation coefficient (PCC) is another popular and
well-documented CM used in drug intelligence [9,10,12,13,17,31–
37,39,54,69]. As with the SCF, the PCC correlation is a measure of
the angle between the two vectors a and b. Hence, the smaller the
angle, the more correlated the values are, with the correlation
determined by:
Fig. 1. Schematic of most common linked vs unlinked population distribution, Xn
adapted from Janzen et al. [45]. ½ðai  aÞðbi  bÞ
rab ¼ qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
Xn
i¼1
Xn
i¼1 i
ða  aÞ2 ðb  bÞ2
i¼1 i
The frequency of scores in both populations are often plotted as a P
histogram. Ideally, there should be no crossover, meaning that the where a ¼ 1n ni¼1 ai ; and analogously for b. The PCC correlation
true positive (TP) and true negative (TN) rates should be 100%. function is quite often modified to scale the results between 0 and
However, it is very common to find that the two populations overlap, 100, this is achieved through the formula:
see Fig. 1. Once the scores of the two populations are elucidated, a ð1  rab Þ
threshold value (THV) is set. This THV allows for the determination of PCC ¼  100
2
linkage for new specimens, e.g. it is possible to assess the level of
similarity between a new specimen and the existing specimen in the
database. Based on the position of the THV, links between the new 3.2.3. Euclidean distance
seizure and the specimens in the existing database can be uncovered. The Euclidean distance (EUC) is another measure of specimen
Information of this nature is beneficial for intelligence-led policing as similarity and is one of the simplest methods used. When two or
it uncovers previously unknown links. more variables are measured, each specimen can be represented by
This position of the THV is dependent on the decision makers a point in an n-dimensional space. When considering points a and
needs and can be altered for different purposes [33]. If a law b, the EUC is the length of the line segment connecting them [90].
enforcement agency is limited to specific resources, the decision The distance between the two points is written as:
vffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
maker might choose to set the THV to minimise false positives u n
uX
EUC ¼ t
(FPs). If the aim is to maximise the intelligence value of the data, 2
ðai  bi Þ
then the value can be shifted to minimise false negatives (FNs). i¼1
It is often the case that large seizures exhibit high intra-
variability [78]. This becomes a significant issue when linked and This method has been utilised extensively for the comparison of
unlinked populations need to be defined. Significant variability in various drug types [11,25,31–33,36,38,40,41,45,68].
the linked population will increase the rate of FNs in subsequent
statistical analyses. For this reason, seizure membership needs to 3.2.4. Manhattan distance
be clearly defined before CMs are attempted. The Manhattan distance (MAN) or taxi-cab geometry gets its
The information generated from applying chemometric tech- name from the perpendicular streets of Manhattan [91]. This grid
niques to forensic case data is often stored in a structured memory layout means that the shortest distance a car could take between
[5]. As the memory is updated, the optimum combination of CM two intersections is equal to the distance calculated by the MAN
and PT needs to be periodically re-evaluated to ensure the most distance. This distance can also be used in cluster analysis (CA) [92]
significant separation between populations is achieved. and is given by the formula:
The subsequent sections outline the theory behind the most Xn
frequently used CMs. In the equations below, ai and bi are the pre- MAN ¼ i¼1
j ai  b i j
treated responses of impurity i, in specimens a and  b,
where the distance between a and b is the sum of the absolute
respectively; a and b are the mean pre-treated peak areas of all difference of their coordinates. Unlike the EUC, which is composed
compounds in specimens a and b, respectively; n is the maximum of one-line segment, the MAN distance contains multiple line
number of compounds present in a specimen; z is the maximum segments.
number of compounds that are zero for specimens a and b.

3.2.1. Square cosine function 3.2.5. Canberra distance

The Squared cosine function (SCF) is a widely utilised and well- The Canberra distance (CAN) is a metric often used for
established method for differentiating between linked and compounds with values close to zero [93]. It first appeared in
unlinked specimen drug populations [9,10,12,31–33,36,68,69]. the literature over 50 years ago and the metric was modified by the
To estimate the similarity of two vectors, the angle between them same authors to present the more commonly used Adkins form of
is calculated. The correlation (SCF) value between the two the Canberra distance [94], its formula is given by:
chromatograms is given by: 1 Xn ðjai  bi jÞ
2 3 CAN ¼
2 nz i¼1 ða þ b Þ
i i
ða b þ a b þ    þ a b Þ
SCF ¼ 100  4 5
1 1 2 2 n n
  2 2 2 The distance is not affected if one of the values in the equation is
a21 þ a22 þ    þ a2n  b1 þ b2 þ    þ bn
zero, i.e. if ai or bi are zero the fraction becomes 1. However, this
In many cases, the modified cosine function is used so that a distance is very sensitive to a small change when both values are
score of 0 indicates a high correlation between specimens and a close to zero. While the MAN and Canberra distances are similar,
6 A. Popovic et al. / Forensic Science International 302 (2019) 109911
the benefit of the latter is that it is not as influenced by compounds
with large values [95].
3.3. Unsupervised pattern recognition
The main goal of these mathematical methods is to correctly
group objects of similar architecture together and different objects
in discrete classes. The concept of class knowledge is what differs
supervised pattern analysis from unsupervised techniques. In
unsupervised pattern recognition (UPR), natural clusters can be
identified in the dataset. The classes are determined through the
application of unsupervised techniques and are not required
initially. In contrast, supervised techniques require the knowledge
of specimen origin to develop models.
Conventional UPR techniques in the field of drug intelligence
include principal component analysis (PCA) and CA. Although
clustering research has its origins in the late 1800s, no widely
accepted theories for clustering exist [96,97]. For this reason,
hundreds of clustering algorithms have been developed over time
(see Fig. 2 for an outline of types of clustering methods). It is not
feasible to outline and explain each algorithm in the scope of this
article. For that reason, clustering algorithms used extensively in the
drug profiling field will be mentioned, i.e. hierarchical clustering
and k-means clustering (KMC). A more exhaustive collection and
explanation of clustering algorithms can be found in [98,99]
3.3.1. Principal component analysis
One limitation of multivariate data is that visualisation of data
with many variables is not feasible. To overcome this issue, some
variables can be excluded through dimensionality reduction. PCA
can achieve this and has been widely utilised in the field of drug
intelligence/profiling [10,11,41,52,53,60,100]. Data dimensionality
is reduced as a result of PCA rotating the coordinate system of the
dataset [101]. The rotation creates uncorrelated variables called
principal components (PC), which are orthogonal and uncorrelated
to each other. This process aims to eliminate high variable
correlation, which in turn would reduce the amount of irrelevant
data from the visualisation or further analysis.
Eigenvalues are a measure of the overall variance of the dataset
for a particular PC. PC scores represent the relationship between
specimens, while PC loadings represent the relationship between
variables and their associated PC.
3.3.2. Hierarchical clustering analysis
One of the most popular ways of grouping objects is hierarchical
cluster analysis (HCA), which presents the clusters in the form of a
Fig. 2. Different types of clustering algorithms.
dendrogram. The y-axis of the dendrogram represents the
similarity/distance between objects, which are plotted on the x-
axis. Different CMs are used to create a similarity or distance
matrix which is used to form the dendrogram. In an ILP context,
HCA is often used to create classes which cluster specimens based
on similarity [5]. For example, if chemical characteristics are used,
specimens are grouped into chemical classes (CCs) if their
similarity exceeds the chosen THV.
HCA dendrograms can be elucidated through agglomerative or
divisive clustering. Agglomerative clustering fuses individual objects
into small groups which are subsequently fused into larger groups
until all objects belong to one cluster. Divisive clustering begins with
a single cluster (containing all specimens) which is divided into
smaller and smaller groups until each object is its own group. There
are many linkage methods which are used to calculate the distance
between clusters and thus influence the structure of the dendro-
gram, (see Fig. 3). These linkage methods include:
a Nearest neighbour, which uses the distance between the two
nearest objects belonging to separate clusters.
b Furthest neighbour, which uses the distance between the two
furthest objects belonging to separate clusters.
c Centroid method, which uses the distance between the centres
of two separate clusters.
d Averaged linkage or group average clustering method, which
calculates the mean distance between all pairs of objects
belonging to separate clusters. The method is intermediate
between the single and complete linkage strategies, thus
attempting to compensate deficiencies of one strategy by the
advantages of the other [102].
e Ward’s method, which computes the distance between clusters
as a minimum of within-cluster variance. The two clusters that
are characterised by the smallest variance between their objects
are merged to build a new cluster.
HCA does not indicate the variables, which contribute most to
the classification of objects in the dataset, unlike PCA, where the
loadings indicate the variation contained within each variable.
There may be a loss of information in the dendrogram generated by
HCA, especially if clusters are not well resolved. However, HCA
does present all the variation in the dataset, in contrast to PCA,
where only a percentage of the variation is typically presented.
3.3.3. K-means clustering
Another popular method used for finding natural clusters in a
dataset is KMC. The goal of this algorithm is to partition all objects
 A. Popovic et al. / Forensic Science International 302 (2019) 109911
Fig. 3. Visualisation of different HCA linkage methods; (a) nearest neighbour,
(b) furthest neighbour, (c) centroid, (d) average and (e) Ward.
into k clusters so that the within-cluster sum of squares (WCSS) is
minimised. In contrast to HCA, the number of clusters, k, needs
to be specified before analysis. Additionally, it is possible that
the lowest WCSS may not be achieved for each cluster as the
technique is optimisation based. This optimisation is achieved
through varying the centroids of the k clusters until no movement
of an object from one cluster to another will minimise the WCSS
[103].
In comparison to HCA, KMC is better suited to larger datasets as
visualisation via dendrogram can get challenging to interpret with
many specimens. One drawback of KMC is that results are
influenced by choice of k. KMC results may differ if the algorithm
is re-run, whereas with HCA, the results are reproducible. In
contrast to HCA, KMC does not produce a graphical representation
of the clustering result. Instead, KMC produces a list of objects that
belong to each cluster. KMC is often paired with PCA to visualise
results or reduce dimensionality before analysis [104].
3.4. Supervised pattern recognition
In supervised pattern recognition (SPR), the number of clusters
is known in advance. Rather than locating natural clusters, SPR
aims to assign unclassified objects to an existing cluster. SPR is
often referred to as classification or discriminant analysis (DA).
Conventional SPR techniques in the field of drug intelligence
include partial least squares – discriminant analysis (PLS-DA)
[12,31,52,53,105], linear discriminant analysis (LDA) [10,71,106–
109] and artificial neural networks (ANN) [60,61,100].
3.4.1. Partial least squares – discriminant analysis
Partial least squares regression (PLSR) is a regression method
based on covariance. PLS-DA is the result of fitting PLSR to DA,
which in turn is a supervised classification method. The PLS
algorithm is used to explain and predict the membership of
specimens to several classes using quantitative or qualitative
explanatory variables. The class membership vector is populated
with “dummy” variables. These variables usually have a value of
either 0 or 1 if they do or do not belong to a class, respectively [52].
During the prediction stage, the closer a specimen is to 1, the more
likely it is that it belongs to that particular class.
In contrast to PCA, which focuses on dimensionality reduction,
PLS-DA is a technique for identifying and measuring variables that
cannot be measured directly (latent variables). Thus, PLS-DA scores
describe the position of each specimen in each determined latent
variable.
7
PLS-DA can be applied in many cases when classical DA cannot
be applied. For example, when the number of observations is low,
and when the number of explanatory variables is high. When there
are missing values, PLS-DA can be applied to the available data.
Additionally, PLS-DA is a type of parametric technique and is
applied to data which is known to have a normal distribution [7].
3.4.2. Linear discriminant analysis
Dimensionality reduction and classification are two common
reasons LDA is applied to a dataset. It is an SPR technique as it
requires knowledge about group membership. The distance
between a specimen and the class centre is calculated and a
new set of axes are calculated. The goal of LDA is to minimise the
distance between specimens of the same class and to maximise the
distance between individual classes.
As LDA requires that the number of specimens exceeds the
number of variables, it is often preceded by PCA [110]. It should
be mentioned that LDA assumes normally distributed data and
common class covariance matrices [111]. However, this is more so
relevant for LDA used as a classifier and not as a dimensionality
reduction technique. In saying that, the technique is still capable of
providing good results even if the assumptions are violated [112].
3.4.3. Artificial neural networks
There have been few articles documenting the successful
classification of illicit drugs using ANNs [60,61,100]. The benefit
of this technique is that ANNs can learn and adapt as opposed to
other chemometric techniques which strive to find the correct
answer [113]. In contrast to PLS-DA, ANNs, which are a type of non-
parametric method are applied to datasets where the distribution
is unknown or known to be not-normal [7]. ANNs are quite useful
for solving non-linear relationships, even in the most complex
datasets [88].
The most common structure consists of neurons arranged into
three different layer types, i.e. input (I), hidden (H), and output (O)
layers, (see Fig. 4). Various types of ANN structures exist with the
multi-layer perceptron (MLP) method featuring most regularly in
the literature [114]. Other types of ANN structures include the
radial basis function (RBF) [115]. MLP usually has one or more
hidden layers, while RBF usually has one. This single hidden layer
allows for faster computation when compared to other methods
[116]. Other ANN structures include probabilistic neural networks
(PNN), which have an additional layer (summation layer) between
the input and hidden layers [117]. Additionally, some articles have
looked at clustering techniques coupled with ANN, for various
applications [118,119].
ANNs tend to learn through the application of training data
which contains defined inputs (e.g. target variables) and outputs
(e.g. chemical classes). Each neuron in the hidden layer receives
its inputs from the training data and summates the product of
inputs and their associated weights. An activation function is
applied to the weighted sum, generating an appropriate output
value [103]. This output serves as the input for the final layer, i.e.
the output layer. The most common activation function is the
sigmoid function, which is used to solve non-linear patterns [103].
As well as the ability to differ the activation function, different
training algorithms can be applied to the ANN. The most common
training algorithm is back-propagation [103]. In this algorithm,
data is fed-forward through the ANN to optimise the weights of
each neuron in the hidden and output layer. This is done to
minimise the error between the actual output and the training
output values [60]. The error in the output layer is then ‘back-
propagated’ to the other neuron layers in the model and weight
values are continuously altered to arrive at the minimum error
[120]. An alternative training algorithm is the conjugated gradient
descent, which is more sophisticated [60].
8 A. Popovic et al. / Forensic Science International 302 (2019) 109911
Fig. 4. Visualisation of different ANNs; (a) radial basis function, (b) multi-layer
perceptron, (c) probability ANN.
4. Evaluation of chemometric techniques
4.1. Evaluation of CMs
4.1.1. Discrimination power
It is vital that statistical methodologies can discriminate
between specimens of differing origins. Thus, calculating the
discriminating power (DP) of the CM is essential. It is through the
calculation of mean and standard deviations of the comparison
scores that the discrimination is calculated [9]. Hence, discrimi-
nation can be estimated by:
mðunlinkedÞ  s ðunlinkedÞ
¼
mðlinkedÞ  s ðlinkedÞ
where m is the average and s is the standard deviation. This
essentially measures the ability of the CM to differentiate between
linked and unlinked specimens. The higher the discrimination, the
better the method is at differentiating unlinked specimens [31]. It
should be noted that the ratio is only an estimation as it assumes
the populations follow a normal distribution, which is often not the
case with casework specimens.
Fig. 5. Sections of a confusion matrix; (a) two-class, (b) three-class.
4.1.2. Confusion matrices
Confusion matrices are a popular evaluation tool for several
chemometric techniques [52]. In the case of combination
evaluation, confusion matrices evaluate the extent of TPs, FPs,
FNs and TNs of the chosen model, (see Fig. 5). The layout of the
matrix is quite intuitive as the instances in the predicted and true
classes are represented in the rows and columns of the matrix.
Using the formulas provided in Table 6, the sensitivity (true
positive rate) and specificity (true negative rate) of a model can be
evaluated. Sensitivity is defined as the probability of getting a
positive result if the specimens were in fact linked. Similarly,
specificity is defined as the probability of getting a negative result if
the specimens were, in fact, unlinked [121]. The closer to one each
of those parameters is the better the model is at classification [53].
This model works just as well for models with more than two
classes (e.g. ANN model), although the structure of the matrix is
slightly altered, (see Fig. 5 for a three-class example).
4.1.3. Receiver operating characteristic curves
Receiver operating characteristic (ROC) curves are another
common way for visualising and assessing the overall efficiency of
statistical combinations [28,122]. This type of curve elucidates the
ideal statistical combination, i.e. the combination with the smallest
intra-variability and greatest inter-variability. As mentioned in
Section 3.2, there will almost always be some overlap between
linked and unlinked populations (see Fig. 1).
For every THV there will be some level of FPs and FNs. ROC
curves are a plot of the sensitivity (true positive rate) as a function
of 1 – specificity (FP rate), see Table 6 for formulas. A test with
perfect separation between the linked and unlinked populations
would have a 100% true positive rate and a 0% FP rate. Therefore,
the closer to 1 the area under the ROC curve (AUC) is the more
efficient the statistical combination [9]. Any model that produces a
curve close to or under the reference line (i.e. 0.5) is a poor
combination of PT and CM. That is, the combination produces a
large overlap between the linked and unlinked populations. A
typical ROC curve would resemble Fig. 6.
Either confusion matrices alone or in combination with ROC
curves can be used to evaluate the ideal statistical combination of
PT and CM.
Table 6
Formulas of certain evaluation technique measures.
Measure Formula
TPþTN
Accuracy
Sensitivity
TPþFPþFNþTN
TP
TPþFN
Specificity TN
FPþTN
 A. Popovic et al. / Forensic Science International 302 (2019) 109911
Fig. 6. Ideal ROC curve example and reference line.
4.2. Evaluation of UPR techniques
4.2.1. PCA
It is possible to determine an ideal number of PCs using several
approaches. These approaches can indicate a different number of
PCs for the same data.
The more variables that load onto a particular PC, the more
critical the PC is in summarising the data. Eigenvalues with a value
of 1.0 indicate that the PC contains the same amount of information
as a single variable. Therefore, the Kaiser method suggests only to
keep PC with eigenvalues above 1.0 [123]. However, there has been
some criticism that this method may allow for too many
components [124].
The Cattell scree test [125] is another method of determining
an ideal amount of PCs to keep. Scree plots are a graphical
representation of eigenvalue magnitude as a function of their
associated PC. They often show a steep curve, followed by a bend
and then a shallow gradient (scree). The ideal number of
eigenvalues will be before the scree, where the line starts to
decrease smoothly.
Each PC accounts for a certain percentage of the variance in a
dataset. The first PC will account for the most variability, followed
by the second and so on. It has been suggested to only retain the
PCs that have a cumulative explanation over a certain percentage.
The percentage chosen is pre-determined and case specific. The
cumulative percent explained method is more subjective than
the Kaiser or scree plot methods.
4.2.2. Cluster validation
Determining the optimal number of clusters in a dataset is
a fundamental issue in partitioning clustering, such as KMC,
requiring the user to specify the number of clusters (k) to be
generated. The number of clusters is somewhat subjective and
depends on the technique and parameters used for clustering.
There are three main methods (elbow, silhouette and gap)
which can be used to determine the optimum number of clusters
[126]. These methods can be used for most clustering techniques.
They will be explored in the following subsections.
4.2.2.1. Elbow method. The elbow method is employed when
attempting to estimate an optimal number of clusters for k means
clustering. Existing methods cannot currently determine the exact
value of k; the elbow method remedies this by looking at the total
9
within-cluster sum of squares (WCSS) as a function of the number
of clusters to provide an accurate estimate. More specifically, the
method works by inserting a range of k values in clustering
algorithms, computing a WCSS for each k value. These values are
used to plot the WCSS curve. The bend in the curve (elbow point)
indicates where the rate of decrease sharply shifts and as such
correlates with an optimal number of clusters [126].
One thing to note with k means clustering is that decreasing the
WCSS increases the distance between clusters. The essence of
partitioning methods, like k means clustering, is to minimise the
WCSS as much as possible as it measures the compactness of
clustering.
4.2.2.2. Average silhouette method. The average silhouette method
is an alternative to the somewhat ambiguous elbow method. It
essentially measures how well objects fit in a cluster and can be
used with any clustering technique [126].
After clustering has been performed, each object is scored a
value from 1 to 1. The closer to 1 the more similar the object is to
its own cluster compared to other clusters. This is repeated for a
range of k values. The estimate of the optimal number of clusters is
found by locating which configuration contains the highest
average silhouette score [127].
4.2.2.3. Gap statistic method. The gap statistic is an approach that
can be applied to any clustering method [128]. It is a formalised
version of the elbow method and employs the standardised
logarithm of the WCSS to calculate the optimum number of
clusters [126].
The idea behind this method was to compare the logarithm of
the WCSS against a distribution with no apparent clustering [129].
Once again, this process is repeated for a range of k values. Ideally,
the clustered specimens will be distant from the randomly
distributed points. The clustering configuration which maximises
the gap statistic is a good estimation of the optimal number of
clusters.
4.3. Evaluation of SPR techniques
Although DA and ANNs differ slightly in functionality, their
resultant models can be evaluated in similar ways. In saying that,
the methods mentioned in this section can evaluate most SPR
techniques. Once an SPR technique has classified new specimens,
cross-validation is often performed to test the classification
accuracy. There are several methods available for cross-validation,
which include resubstitution, hold-out method and the leave-one-
out method [110]. As mentioned in Sections 4.1.2 and 4.1.3,
confusion matrices and ROC curves are not just suited to CMs. They
have been used in many cases to evaluate SPR techniques including
PLS-DA, LDA and ANN models.
Additionally, “predicted vs measured” plots can be used to
evaluate the quality of the regression model fitted to the data [31].
As the name suggests, these plots assess the correlation between
the predicted values and actual values of a specimen. The closer the
correlation between the two axes the better the PLS-DA or ANN
model is at predicting classes. This type of plot visualises the
prediction error for calibration and validation datasets. Other
methods of evaluating prediction error have been outlined in
[130,131].
Apart from the methods mentioned above, PLS-DA and LDA
models can be evaluated via several graphical methods. These
include score plots or loading plots [12].
DA score plots vary to that of PCA due to the supervised nature
of the former technique. While PCA transforms a dataset based on
the greatest orthogonal variation in the data. DA, on the other
hand, produces a similar transformation but is informed by
10 A. Popovic et al. / Forensic Science International 302 (2019) 109911

between-group variability to better reveal group structure
[132,133]. DA loading plots can be interpreted similarly to PCA
loading plots. Highly correlated variables have similar weights in
the loading vectors and appear close together in the loading plots
of all dimensions.
5. Application and added value of chemometric techniques
5.1. Applications of CMs to illicit drugs
The combination of PT and CM will influence the rate of FPs and
FNs obtained for a particular dataset. Subsequently, the selection of
the THV can be modified to reflect the appetite for risk, resource
availability or other operational factors.
Table 7 outlines various statistical combinations found in the
literature, referring to a variety of illicit drug types. As mentioned
in previous sections, the optimum combination will ensure the
greatest separation between linked and unlinked populations,
which in turn minimises FP and FN specimen linkages. In some
cases, authors have chosen to focus on one statistical combination
rather than comparing modifications. For simplicity, it should be
noted that the optimum or only statistical methodology in the
respective reference is in bold. The remainder of the references
present in Table 7 is a combination of lower ranked options, which
the reader should not be discouraged to try as the effectiveness of
the lower ranked options is not significantly different. The
combinations have been ranked based on the DP or the AUC for
the ROC curves stated in the article, either-or is present in the
referenced articles.
The following sections break down the preferred and optimum
statistical combinations for each illicit drug type, as per the
available literature.
5.1.1. ATS (AM & MA)
For amphetamine (AM) and MA, the EUC is the most widely
utilised metric for determining the linkage between specimens of
AM and MA. This may reflect the ease of computation and familiar
nature of this method. However, in the cases where authors have
experimented with metrics other than the EUC, it generally does
not produce the greatest separation between populations. In terms
of statistical combinations, N + 4R/PCC and N + 4R/SCF consistently
produced lower FP rates for a variety of studies.
Four articles focused their efforts on using only EUC as a
distance measure [25,38,40,41]. This CM showed satisfactory
discrimination between the linked and unlinked populations.
Dujourdy et al. [12] evaluated EUC, SCF and PCC and determined
that the two latter metrics provided exceptional discrimination
between the two populations, with SCF producing a slightly lower
FP rate. This result was also supported by further studies [32,36]
with SCF the most effective statistical combination when
compared against other metrics.
When multiple PTs were compared, N + 4R or N + 2R produced
the best discrimination between populations [12,31,32]. Ander-
sson et al. [31] theorised that these PTs are more discriminatory
than N as they minimise closure effects. Closure, or the constant
sum, introduces dependence between normalised variables so that
if one large variable goes up, the others automatically go down
because their sum is fixed [134]. Using a treatment such as N + 4R
reduces the influence of large peaks and classify data with more
accuracy, thus minimise closure effects.
5.1.2. ATS (MDMA)
In many cases involving MDMA, PCC with data PT using
normalisation and its modifications, namely N + 4R, has been found
to produce the greatest separation between linked and unlinked

Table 7
Statistical combinations of CMs and PTs used for drug profiling, references shown in cells. Where applicable, the optimal CM for the article is shown in bold. The second and
third ranked combinations in the relevant articles are included as well. Any article that has evaluated only one combination is italicised.

CM PT ATS (AM & MA) ATS (MDMA) Cocaine Heroin

SCF L [36]
N [37] [9] h , [47], [136] [69], [68] c
N+L [56]
N + 2R [12] [10]
N + 4R [31], [12], [32]a , [32]b [33]a , [33]b
N+S [9]g , [9] h
N + S+L [56]
PCC 4R [56]
L [36]
N [10], [35] [54] [69]
N+L [35]
N + 2R [12] [39], [10], [34] e [17] [17]
N + 4R [31], [32]a,b [33] , [33]a , [35], [34] f
b

h g
N+S [9] , [9] [13]
EUC 2R [36]
4R [11]
N [45] [68] d
N + 4R [31], [32] b, [32]a [33]a,b
S [41], [25], [40], [38] [11]
CAN N [69]
N+L [9]g
N + 4R [31]
MAN 4R [11]
S [11]
a
Organic impurities.
b
Inorganic impurities.
c
Major impurities.
d
Minor impurities.
e
LLE method.
f
HS-SPME method.
g
Within-laboratory study.
h
Between-laboratory study.
 A. Popovic et al. / Forensic Science International 302 (2019) 109911
populations [10,33–35,37,39]. It is always good, if time permits, to
explore all combinations of CMs (including their modifications)
and PTs.
Morelato et al. [33] applied PCC, SCF and EUC metrics to
inorganic impurity data, with PCC found to produce the greatest
separation of linked and unlinked populations and the lowest FP
rates. This outcome was also observed for additional studies,
although for organic impurities, where multiple metrics were
compared [34,35]. In another study by van Deursen et al. [39], PCC
was evaluated as the only method. Although a good separation of
populations was achieved, the authors concluded that a larger
specimen set was needed to increase the accuracy of their analyses.
Additionally, in their final remarks, it was suggested that
alternative statistical combinations should also be trialed. This is
unsurprising as different metrics can have varying effects on the
structure of the linked and unlinked score distributions.
In contrast, two studies [10,33] utilising MDMA organic
impurities concluded that the use of SCF, rather than other
metrics, produced slightly lower FP values for both datasets. An
additional study by Gimeno et al. [37] presented the SCF as a means
of possible identification of batch-level relationships based on
MDMA tablet impurities. However, the profiling technique needs
to be applied to more MDMA specimens to set an accurate THV for
distinguishing populations. In the sequential study to Weyermann
et al. [10], Marquis et al. [11] evaluated EUC and MAN distances for
the diameter, thickness, weight and score of seized MDMA tablets.
In this case, it was found that EUC resulted in a separation of linked
and unlinked populations. One reason for this might be that the
EUC tends to perform better than MAN when the dataset contains a
low number of outliers [135].
The greatest separation between populations in most cases
was achieved when using the N + 2R and N + 4R PT method [10,33–
35,39]. It should be noted that studies [11] and [37] did not
evaluate the performance of N + 2R or N + 4R.
5.1.3. Cocaine
Although cocaine profiling and the subsequent generation of
intelligence products have been documented in the literature
[9,17,45,47,54,56], the summary presented in Table 7 depicts the
lack of versatility of statistical methods used in the articles above.
Only articles [9] and [56] explored more than one CM, and in
both cases, SCF and PCC outperformed other metrics. In the case of
[9] where a cross-border inter-laboratory specimen comparison
was attempted, it was found that the same metrics produced the
greatest discrimination between populations. Two articles [17,54]
evaluating PCC only, reported that a small overlap of the linked and
unlinked score distributions was observed. Broséus et al. [54]
noted that this degree of overlap is reliant on sampling and the
statistical combination should be evaluated using a larger
specimen set. Nonetheless, PCC has proven to be a viable CM for
the differentiation of cocaine specimens. In contrast [136], and [47]
reported minimal FP and FN rates when using SCF with N. It is
interesting to note that Dujourdy and Besacier [47] tested whether
the addition of cocaine base would influence the similarity
measurement. This was done to test the applicability of the
Table 8
Common unsupervised and supervised pattern recognition techniques split by drug type.
Pattern recognition technique ATS
UPR PCA [10,11,41,100]
HCA [12,25,36,41,100]
KMC
SPR PLS-DA [12,31]
LDA [10]
ANN [100]
11
profiling method to specimens which have not properly crystal-
lised and thus half base half HCl.
As [17,45,47,54,136] only looked at one PT it is difficult to report
which is commonly the optimum. Lociciro et al. [9] determined
that out of 66 statistical combinations, N + S paired with SCF and
PCC provided optimum results. Liu et al. [56] evaluated 50
combinations and reported that N + S + L proved to have the
greatest DP. Interestingly, S + N tended to have a very low DP for all
CMs.
5.1.4. Heroin
Critically evaluating an optimum statistical combination is
difficult at this stage due to the lack of diversity in combinations
applied in the few articles that exist for heroin profiling.
In general, few studies apply all five metrics mentioned in this
review article to a single dataset. One such study by Esseiva et al.
[69] found that there was an overlap between linked and unlinked
populations, no matter which statistical combination was used.
Their findings showed that the EUC and MAN, in comparison to
CAN, SCF and PCC, showed a larger distribution of linked scores and
were therefore excluded from further analysis. Although only one
PT was explored, N, its combination with SCF proved to produce the
lowest rate of FPs [69]. Dufey et al. [68] investigated the use of both
minor and major impurities for determining links between heroin
profiles. In another interesting article, Broséus et al. [13] compared
the performance of PCC on its ability to separate linked from
unlinked specimens within each analytical method and between
analytical methods. Although the methodology was applicable for
their aims, the authors suggest that further exploration of
statistical combinations should be conducted. This type of research
is at the forefront of developing solutions to the issue of method
harmonisation.
In contrast to other drug types, a search of the literature shows a
small selection of PTs utilised for heroin data. It is highly
recommended that other PTs are explored for a thorough
evaluation of the optimum statistical combination.
5.2. Applications of UPR to illicit drugs
5.2.1. ATS (AM, MA & MDMA)
The visualisation and identification of clusters in ATS datasets
has been achieved in several studies which employed PCA and
HCA, see Table 8. Overall, PCA provided suboptimal results when
utilised for pattern recognition of ATS specimens. Although this
was the case in several articles [10,11,41,100], the method should
not be overlooked as it was successful for other drug types, as
explained in the subsequent sections. Furthermore, except for one
study [100] by Waddell et al., HCA successfully recognised known
and identified new patterns in several datasets [12,25,36,41].
Several articles utilised PCA to group specimens according to
their original seizure [41,100] and country of origin [10,11].
Suboptimal results were reported for all four articles as the
structure of the groups did not coincide with additional specimen
information. It is not guaranteed that specimens coming from the
same seizure are linked; this is only an assumption. For this reason,
Cocaine Heroin
[52,53,107,139,141,142] [60,61,66,71–76,143]
[47,52,53,56,136,138,140,142] [66,67,71,73,75,144]
[61,67,71,75,143]
[47,52,53,105,138,139] [74]
[106,107] [71,108,109]
[60,61,66]
12 A. Popovic et al. / Forensic Science International 302 (2019) 109911
specimen membership to pre-classified groups needs to be
carefully evaluated via visual inspection of score plots. That is,
specimens logged as belonging to the same seizure should be
carefully evaluated before attempting a pattern recognition
technique.
Furthermore, PCA was unable to detect patterns between
countries, likely due to the production of MDMA utilising
predominantly the same route (reductive amination) [10,11].
The variation in MDMA tablets can be attributed to the high seizure
intra-variability in both studies. In both cases, the authors
recommended that statistical analysis should be applied to
specimens collected in similar periods to arrive at more reliable
conclusions about patterns in the data.
In contrast, most articles involving HCA reported successful
results [12,25,36,41] except [100]. Despite indicating the potential
grouping of tablets, not one original seizure was correctly clustered
by HCA in the latter article [100]. The ability of the technique to
correctly group seizures was limited by the high intra-variability of
the specimens. It is difficult to overcome this issue as MDMA
tablets produced clandestinely will almost always exhibit such
variability [100].
However, in the case of studies [41] and [36], HCA was able to
group specimens into their original seizures except for a few
misclassifications. Krawczyk et al. [41] used EUC and PCC as
distance measures, and the single linkage method to determine
cluster structure. Upon viewing the resultant dendrograms, it was
elucidated that HCA was an adequate technique to evaluate class
membership. The visual nature of the dendrogram allows analysts
to easily set a THV as to whether specimens belong to the same
group or not. The score on the y-axis of the dendrogram can be
linked to certain FP and FN values, which can then be used to set an
appropriate THV [56]. Although the analysis performed by
Kuwayama et al. [36] also contained minimal error, it is important
to factor in the small specimen size. To ensure the accuracy of the
technique, the specimen size should be increased.
Both Dujourdy et al. [12] and Kuwayama et al. [25] identified
interesting trends in their respective MA datasets using HCA. The
former study performed HCA on a group of MA specimens
previously determined as being produced via ephedrine (EPH),
(achieved through PLS-DA, see Section 5.3.1). However, MA
synthesised via EPH can be achieved in a variety of ways, e.g.
through Birch reduction, Nagai and Moscow routes [137]. Distance
calculation between specimens via EUC paired with the Ward
linkage method identified the presence of six clusters indicative of
different synthesis routes. Furthermore, it was possible to indicate
which target compounds were characteristic of each cluster,
yielding information about the applicability of the chosen target
compounds. In the latter study by Kuwayama et al. [25], authors
clustered MA crystals from different countries based on charac-
teristic impurity peaks. These clusters provided information about
tendencies of specimens in different countries, in particular, the
lack of naphthalene derivatives present in Thai MA specimens.
5.2.2. Cocaine
As cocaine mainly exists in two forms, i.e. as the salt and base, it
is essential to be able to differentiate between these two chemical
forms as they indicate the extent of the cutting process
[52,53,138,139]. An alternate use of UPR can be seen in
[47,56,107,140] and [141,142]. The aim of these articles was the
elucidation of common geographical origin and batch trends,
respectively.
In [52,53,139], score plots of the first two PCs graphically
display the separation between different cocaine forms. In the case
of Rodrigues et al. [52] three main groups were observed; high
purity cocaine HCl, high purity cocaine base and low purity cocaine
(both HCl and base). The PCA conducted by Marcelo et al. [53] was
not as definitive but indicated a separation between cocaine HCl
and base, with the purity and therefore potential adulterant
presence to be inferred by the wide range of each group. The latter
article [139] elucidated that cocaine specimens of various purities
could be differentiated via PCA. Additionally, using loading plots,
authors were able to infer which impurity peaks were responsible
for the discrimination between specimens.
Monfreda et al. [107] and Nielsen et al. [141] obtained alkaloid
and residual solvent profiles for cocaine seizures. In both cases, the
alkaloid profile was more descriptive of the seizure subgroups than
the solvent profile. It was hypothesised that due to the lack of
variation in latter profiles, multiple batches were manufactured
using the same organic solvent [141]. Combining the two profiles
did not yield more information than the alkaloid PCA alone. Rather
than combining them, it was suggested that the two profiles could
be complementary. Production batches, identified through alka-
loid profiles, could be linked to the same manufacturer by the
presence of common residual solvents [141].
The HCA analysis seen in [52,53,138] shows that specimens
were clustered by chemical form and purity level. This confirms the
analysis conducted by PCA, where applicable. HCA performed by
Rodrigues et al. [52] and Perez et al. [138] predominantly grouped
the specimens by high and low purity. Marcelo et al. [53] opted to
group specimens into more definitive clusters. That is, HCA results
indicated that specimens were split into two major groups (HCl
and base), and subsequent groups were formed based on the type
of adulterant present. With some misclassification, HCA separated
all specimens based on assumed geographic location
[47,56,140,142]. This analysis allowed for the identification of
previously unknown links between specimens. Additionally, the
purity level could be attributed to certain locations, highlighting
the extent of adulterants used in various regions [140]. Further-
more [136], used HCA as a preliminary clustering technique to
group similar specimens prior to PT/CM analysis. This use of the
technique does not feature greatly in the literature but is useful
when the specimen to seizure relationship is not clear or known.
5.2.3. Heroin
A range of UPR methods has been applied to heroin sets with
the purpose of batch comparisons [60,66,71,73,143,144] and
geographical origin determination [61,67,72,74–76], using either
organic or inorganic components of the heroin production
process.
A clear trend across the range of heroin profiling projects was
the exploration of different combinations and sequencing of
techniques. Overall, PCA was demonstrated to be effective as a
preliminary screening technique [60,61,66,73,75,76,143]. Howev-
er, the technique does not perform as well for low purity specimens
[71]. Some studies identified that combinations of variables are
present in specific concentrations in heroin coming from different
origins [74]. This notion allowed for unsupervised classification of
heroin specimens using PCA score and loading plots [72,74].
However, the specimens were split into two broad origins.
Additionally, it is not guaranteed that the variables were present
at the point of plant cultivation, they could have been added at any
point of the manufacturing process. It would, however, be
interesting to assess the applicability of PCA to classify specimens
into sub-groups and therefore more specific origins.
As mentioned, many authors reported a sequential application of
an UPR clustering technique such as HCA or KMC following PCA. HCA
was used to evaluate the clustering of specimens [66,67,71,73,75,144],
with some authors also conducting a thorough comparison of different
combinations of linkage methods and CMs [67,71]. These studies
identified that the Ward linkage method consistently outperformed all
other options including average, centroid and median; when used in
combination with either squared EUC or squared PCC CMs.
 A. Popovic et al. / Forensic Science International 302 (2019) 109911
The final UPR method to be featured in research involving
heroin profiling data is KMC [61,67,75,143], which can also be
applied as a semi-supervised pattern recognition method, as
demonstrated by [71]. It is interesting to note that contrary to the
articles mentioned above, one article did not use KMC as a
confirmatory technique [67]. Instead, it was used as a dimension-
ality reduction technique, with HCA confirming the results.
5.3. Applications of SPR to illicit drugs
5.3.1. ATS (AM, MA & MDMA)
As a plethora of possible manufacturing routes exists for synthetic
ATS [137], two studies [12,31] sought the use of PLS-DA to extract
information regarding synthesis routes for their respective datasets.
Andersson et al. [31] used PLS-DA to evaluate the optimum PT for AM
route discrimination, in particular, the Leuckart, nitrostyrene and
reductive amination routes. While Dujourdy et al. [12] used the same
technique to discriminate between MA manufactured via benzyl
methyl ketone (BMK) and EPH.
Evaluation of the PLS-DA models was carried out using score plots
and “predicted vs measured” plots. Score plots, in both cases, gave an
indication of the model with optimum discrimination. In the case of
one article by Dujourdy et al. [12], score plots graphically illustrated
that the model successfully differentiated specimens synthesised
using BMK and EPH routes. It also gave an indication of the large
extent of EPH manufacturing, shown by the sizable difference
between the two groups. In the case of [31] by Andersson et al., visual
inspection of the score plots indicated that the N + 4R PT provided the
greatest spread between synthetic route groups.
As visual inspection can be quite subjective, the results of the
score plots were confirmed through “predicted vs. measured” plots.
Dujourdy et al. [12] elucidated that their model produced a very high
correlation coefficient for both calibration and validation, demon-
strating the applicability of their model in discriminating between
MA synthesised via BMK and EPH. The authors also used their
validated model to make predictions with new unclassified speci-
mens, which resulted in a 5% misclassification rate. Andersson et al.
[31], on the other hand, reported that N + 4R produced the highest
correlation coefficient for the “predicted vs. measured” plots.
An alternative DA applied to ATS data was explored by
Weyermann et al. [10]. In this study, LDA was applied to MDMA
data to ascertain whether a reduction of target variables offered
the same level of discrimination. It was found that the model
employing the reduced set of variables presented minor classifi-
cation. Furthermore, the model was also able to identify the
variables mainly responsible for specimen discrimination.
With the limited literature on ANN development for ATS,
Waddell et al. [100] provided an interesting insight into the topic.
The authors classified ecstasy tablets into their original seizures
using various MLP models and retained the six optimum ANN
configurations. These configurations produced an accurate classi-
fication rate of 96–99%. Misclassification can become a major issue
if seizure intra-variability is high. However, this type of variation
will always be present in clandestinely produced illicit substances
[100]. It is a true test of the ANN to be able to discriminate between
linked and unlinked specimens.
5.3.2. Cocaine
As mentioned in Section 5.2.2, cocaine can exist in several
forms. For this reason, it is useful to be able to differentiate
between these forms. Several articles [47,52,53,105,138,139] have
evaluated the use of PLS-DA for classifying cocaine specimens.
Dujourdy et al. [47] attributed certain solvents to three
presumed specimen origins (Peru, Bolivia and Columbia) with
the aid of a PLS-DA loading plot. Rodrigues et al. [52] developed
two PLS-DA models which successfully discriminated between
13
cocaine specimens of low and high purity and specimens of HCl
and base form. Although score plots were not presented in the
article, authors presented confusion matrices, which summarised
the results. The models assessing purity and chemical form
produced sensitivity and specificity in the range of 83–97%. The
models developed by Marcelo et al. [53] and Grobério et al. [105]
each correctly identified 100% of specimens as being either cocaine
HCl or base. In the case of [105] by Groberio et al., the lack of error
was achieved after outliers (4.4% of the dataset) were removed.
Two articles utilised LDA for the classification of cocaine
specimens [106,107]. In the former study [106], specimens were
subjected to different storage conditions and their alkaloid and
residual solvent profiles were generated over time. LDA predicted
the level of association between specimens. The authors suggested
improving the statistical comparison of cocaine profiling by
combining the two profiles. The stability of residual solvent
profiles over time, as well as the specificity of alkaloid profiles,
proved to lower prediction error rates. In the latter article [107],
LDA was modelled on pure cocaine specimens. Test “cut” speci-
mens (containing cutting agents) were classified according to
clusters observed through PCA. In both cases, prediction error only
started occurring when specimens contained more than one
cutting agent and a cocaine percentage of less than 60%.
5.3.3. Heroin
A limited range of SPR methods has been applied to heroin
profiling data, with the typical objective of achieving geographical
origin determination [61,71,74,109]. For heroin profiling, ANN
architectures such as RBF and MLP have been featured several
times in the literature [60,61], with Ratle et al. [61] evaluating
alternative ANN such as PNN and an additional non-parametric
method k-ANN. PNN and k-ANN outperformed the MLP and RBF
methods, but this outcome may be attributable to the application
of a dimensionality reduction technique such as PCA. However,
when discriminating between heroin batches, Klemenc [66] found
that PCA and HCA were less erroneous than k-ANN. It should be
noted that this misclassification by the SPR technique could be
attributed to the small specimen size.
Another SPR technique applied to heroin profiling data is DA.
The technique can be used to discriminate specimens based on
geographic origin [71,74,109] or batch trends [108]. Additionally,
DA was evaluated in comparison to other chemometrics [71,74]
where it was identified as the superior method. In all four cases,
DA performed well with some misclassification. Amongst other
things, the error could be due to the small specimen size,
uncertainty of the country of origin or period of the specimens.
As DA learns from available specimens, a small dataset could
potentially overtrain the model and misclassify specimens
[71,108]. Furthermore, if there is a high inaccuracy in the original
assumption of the country of origin, this could increase the
misclassification rate during the training of a model [109].
With regards to spatiotemporal trend analysis, it would be
beneficial to build SPR models that can discriminate between
different batches with the same geographic origin. For this to be
achieved the outputs of models should not be restricted to the
assumed country but should include subgroups to account for
the change in the manufacture of heroin specimens [109]. Overall,
there would appear to be a significant opportunity to explore SPR
as the bulk of research in this space is occurred over a decade ago.
5.4. Added value of chemometric techniques applied to illicit drugs
for ILP
The sections above provide a consolidated review on the
application of the most common chemometric techniques con-
cerning drug profiling. The subsequent sections go through the
14 A. Popovic et al. / Forensic Science International 302 (2019) 109911
added value of chemometric techniques applied to illicit drugs for
ILP, in particular, their added value towards active investigations
and understanding drug markets. In an ILP perspective, forensic
case data is usually stored in an organised memory [5]. This
includes profile(s), scores, link values relating to specimens [77].
Previously unknown connections between specimens can be un-
earthed through applying chemometric techniques [36,41]. This is
often achieved through a combination of CMs and HCA to create
classes of specimens based on similarity, which are subsequently
integrated into the memory [60]. Classes that are produced
through clustering can be chemical or physical, depending on the
selected characteristics. The memory needs to be updated conti-
nuously to provide accurate information in a forensic intelligence
application, this includes CCs as well as any investigative
information.
5.4.1. Added value for active investigations
If the subset of specimens to be explored is restricted to a
chosen event, chemometric techniques can be used to support a
hypothesised link based on other investigative information. This
approach is rather reactive as specimens to be compared are
restricted to certain cases where links are inferred by other
information. Granted, the hypothesised links may be confirmed or
refuted by the chosen statistical measure, but the potential to
uncover links that are not indicated by investigative information is
severely limited. This process is somewhat restrictive as it severely
confines the potential of drug profiling to support ILP.
Another approach would be to systematically compare new
specimens against existing cases with the aid of chemometric
techniques. The physical result of systematically applying CMs to
drug profiling data could take the form of a record of linked cases
(including previously unknown links). Further applying CA could
index specimens based on their similarity, in turn creating classes.
This would produce a form of intelligence that generates a clearer
insight into the connectivity of the specimens in question to the
drug market. This insight could allow investigators to focus their
resources on exploring instances where specimens are seemingly
linked to unrelated cases. Ideally, these instances should be
supported by other types of information. Furthermore, the process
used to arrive at the link needs to be continuously evaluated to
ensure an appropriate level of FPs and FNs.
An example of both approach was demonstrated by Esseiva
et al. [60]. In this article, several heroin seizures were chemically
profiled for the purpose of confirming and potentially elucidating
further links. Four clusters of specimens, seemingly linked through
investigative information, were further examined through chemi-
cal profiling and chemometrics. The hypothesised links were all
chemically confirmed. If the analysis had stopped there (as it is
traditionally the case), investigators would have missed links with
other seizures not initially connected to the particular investiga-
tion. However, the chemical profiles were compared to all other
specimens in the memory and new connections were revealed that
were previously hidden when consulting investigative information
only [60]. This approach made visible unsuspected links and
provided a more complete picture of the extent of the market.
5.4.2. Added value for understanding drug markets
Chemometric techniques are a valuable tool for supporting ILP;
the approach outlined above could not be achieved by presenting
raw data to investigators. So, as well as systematically comparing
new specimens against the memory, it can be explored retrospec-
tively to understand drug markets and ultimately predict trends
that could prevent future events from happening [145,146]. To
have further confidence in the links and CCs identified through
chemometric techniques, they should be validated with other
sources of information [147,148].
The creation of CCs is quite a useful feature in an ILP context.
Their creation allows us to infer that specimens part of the same
class were once part of the same physical unit, when major
impurities were profiled [16]. Taking the example of Broséus et al.
[16], a drug profiling method coupled with chemometric
techniques was used to study the evolution of cocaine and heroin
drug markets as well as devise measures for promptly disrupting
developing segments of the network. The findings of such analysis
included the degree of drug homogeneity, spatiotemporal market
trends and expected outcomes of preventative measures.
In more detail, the degree of drug homogeneity can be inferred
from mapping the connections between specimens. A network
with a low number of CCs and a high number of connections
indicates consistency amongst specimens [16]. Additionally, hubs
in the network can be identified by observing seizures that link
previously unconnected cases. These instances can be confirmed
through quantitative measures of social network analysis, such as
centrality scores [149]. Based on the centrality scores and number
of CCs per seizure, Broseus et al. [16] showed that cocaine multi-
profile seizures are commonly observed at an upper level of the
distribution chain while this is not the case for heroin. This may
highlight how physical units with different chemical profiles are
managed and possibly blended along the distribution chain, an
information difficult to obtain through traditional market analysis.
Furthermore, spatiotemporal analysis can be performed on
the network to identify points of interest or a need for trans-
jurisdictional collaboration. Using static visualisations may limit the
intelligence that can be generated from network evaluation.
Therefore, it is recommended that the network be studied over a
chosen time (e.g. trimester) to understand the evolution of the drug
market fully. Temporal analysis can elucidate information about CCs
life and infer the emergence or extinction of criminal networks.
Classes deemed significant in size and observed to be emerging
deserve more focus from investigators and should be neutralised
promptly. As all information regarding a specimen is not always
available, SPR can be used to classify entities to gain a better
understanding of markets. There have been many instances of
successful classification, covered in Section 5.3. These articles mainly
classified specimens and highlighted patterns based on geographic
location, route of synthesis or batch trends. As well as temporal
analysis and enrichment of the memory, intelligence can be gathered
from evaluating spatial data. Trans-jurisdictional connections can
identify problematic areas and identify instances where collabora-
tion between jurisdictions should be increased [16]. Additionally, at
an international level, network analysis could identify countries that
are pivotal to drug markets. This could aid the suggestion of strategic
targets for disrupting international drug trade [145].
It is possible to receive an indicator of how well implemented
strategies are disrupting drug markets by analysing the extent
of certain CCs at a given time. To expand on this idea, law
enforcement could focus their resources on CCs deemed to be
important. This would initially result in an increase in connections
within the network as more seizures are made, then ideally a
decrease in connections as the criminal activity has been quenched
due to the focused efforts of law enforcement. However, this isn’t
always the case, criminal organisations adapt to police efforts and
not all attempts at market disruption are successful. That is not to
say that something cannot be gained from this. These unsuccessful
efforts can guide law enforcement in understanding flexibility of
criminal organisations and help further develop proactive strate-
gies for ILP [16].
6. Conclusions
The illicit drug trade is a widely recognised social and judicial
problem. However, there are opportunities to aid decision makers
 A. Popovic et al. / Forensic Science International 302 (2019) 109911
in disrupting the problem through the collection and appropriate
exploration of forensic case data. Chemical and physical profiles,
coupled with chemometric techniques, can be used to highlight
various patterns in the forensic case data. In the case of illicit drug
data, these patterns are related to batch trends, links between
specimens, geographic location or route of synthesis.
The most common chemometric techniques outlined in the
literature include CMs, unsupervised and supervised pattern
recognition.
CMs can be very useful in determining the level of association
between profiles. Multiple metrics should be applied to any dataset
to identify the optimum. The EUC distance has been widely used
for determining linkages as it produces excellent results. However,
when other metrics are explored alongside EUC, it proves to be
suboptimal. These cases show that either SCF or PCC provide the
lowest rate of FPs and FNs. In all the articles included in this review
either SCF, PCC or EUC were superior to other metrics. The optimal
combination of CM and PT needs to be periodically re-evaluated to
accommodate the addition of new specimens to the database. This
will ensure that maximum separation is achieved between linked
and unlinked populations.
The UPR techniques featured in this article include PCA, HCA
and KMC. The occasional error encountered with PCA was most
likely due to incorrect assumptions relating to specimen member-
ship. Otherwise, the technique was successful in preliminary
screening and dimensionality reduction. The two clustering
techniques (HCA and KMC) explored in this article highlighted
interesting patterns between specimens. The UPR techniques are
commonly used sequentially and often to confirm PCA results. The
visual nature of the HCA dendrogram could explain the popularity
of the technique. Significantly fewer articles feature KMC. The two
clustering techniques are not without their drawbacks and the goal
of the data analysis should be considered before choosing one or
the other. Furthermore, one limitation of the clustering techniques
is the requirement of choosing a THV, whether that be a score
relating to a CM or the number of desired clusters. This THV might
not always be an obvious choice and if chosen incorrectly, could
severely limit the information acquired from the clustering
techniques. Additionally, unlike PCA, HCA does not indicate which
variables contribute most to the classification of objects in the
dataset.
When complementary specimen information is available, SPR
techniques can be used to generate prediction models. When
compared to UPR, superior results were often observed for DA.
However, misclassification was encountered in some cases. This
error was usually due to small specimen size, the uncertainty of the
country of origin or lengthy time span of specimens. Several ANNs
have been applied to illicit drug data in order to classify specimens.
In most cases, the ANN produce sufficient results, but the efficacy
may vary if unsupervised techniques are applied first.
The optimum ordering of chemometric methods will depend on
the desired goal. In many cases, PCA is a good starting point. It
can be followed by CA or SPR depending on the availability of
additional information. A prediction model can be generated
without additional class information if CA is performed ahead of
SPR. However, CA can also follow SPR if a known class is thought to
contain further clusters.
Various configurations of chemometric techniques can aid the
interpretation of forensic case data, in particular, their ability to
add value towards active investigations and understanding drug
markets. Drug profiling has proven useful in confirming links
originally hypothesised by investigators and has also demonstrat-
ed the potential of going a step further and identifying previously
unconnected entities. Furthermore, several articles have demon-
strated the ability of forensic case data, coupled with chemometric
techniques, to support ILP in a strategic context. This was achieved
15
through studying the evolution of drug markets, devising
measures for prompt disruption and thorough evaluation of such
measures to ensure success of police efforts.
CRediT authorship contribution statement
Ana Popovic: Conceptualization, Data curation, Writing -
original draft, Writing - review & editing. Marie Morelato: Writing
- original draft, Writing - review & editing. Claude Roux: Writing -
review & editing. Alison Beavis: Conceptualization, Writing -
original draft, Writing - review & editing.
Acknowledgements
This work is supported by the Australian Research Council grant
(LP160100352). The authors would like to acknowledge the project
partners: NSW Forensic & Analytical Science Services, Australian
Federal Police, United Nations Office on Drugs and Crime,
Univertity of Lausanne.
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