Forensic Chemistry 18 (2020) 100223

Contents lists available at ScienceDirect

Forensic Chemistry
journal homepage: www.elsevier.com/locate/forc

Scientific foundations and current state of trace evidence—A review T

a b c,⁎
Tatiana Trejos , Sandra Koch , Andria Mehltretter
a
Department of Forensic and Investigative Science, West Virginia University, Morgantown, WV 26505, United States
b
McCrone Associates, 850 Pasquinelli Drive, Westmont, IL 60559, United States
c
Laboratory Division, Federal Bureau of Investigation, 2501 Investigation Parkway, Quantico, VA 22135, United States

HIGHLIGHTS

• ABibliography
review including scientific foundations and current challenges for trace evidence.
• Emphasis on hair,
serves as support for trace evidence examinations and interpretation.
• fiber, tape, paint, and glass evidence.

ARTICLE INFO ABSTRACT

Keywords: In this paper, a review of the literature concerning the scientific foundations of trace evidence analysis is pre-
Hair sented along with a discussion regarding the current challenges for the discipline. Information about the com-
Fiber position, manufacture, transfer, and persistence of trace materials are discussed with a focus on how each aspect
Tape impacts trace evidence interpretations. This paper reviews the literature that describes the underlying validity of
Paint
trace evidence examination methods and interpretive approaches, highlighting experimental studies that de-
Glass
scribe performance measures, capabilities, and limitations of five trace subdisciplines: hair, fiber, tape, paint,
and glass. This review is intended to serve as a foundational document in support of the trace evidence discipline
and the interpretation approach being developed by the OSAC Materials (Trace) Subcommittee.

1. Introduction
The term “trace evidence” refers to physical evidence that is small
and transfers as a result of interaction between objects and individuals
[1]. These materials are often microscopic in size but can yield in-
formation about what may have occurred, where the sample may have
come from, and how the materials may have transferred. The potential
for trace evidence to play a major role in criminal investigations was
first recognized in the early twentieth century by Edmond Locard, a
French lawyer and medical doctor, who believed that the examination
of trace materials could link places, people, and objects to a crime.
Locard’s exchange principle, often quoted as “every contact leaves a
trace,” soon became a maxim of forensic science [1,2].
Trace evidence examiners use a variety of analytical tools to locate
and examine evidence, identify their main components, classify the
materials, and compare items to determine if they could share a
common source. As Locard predicted, trace evidence is often used to
relate what may have occurred during a crime based on materials found
at a scene. For instance, hairs recovered on a victim’s clothing can
support the proposition of contact with the perpetrator. Tape, and any
debris found adhering to its adhesive surface (e.g., hairs, fibers) can be
used to link people or scenes, to each other or to a potential source
material (e.g., textile garment, carpet, tape roll). In hit-and-runs, the
analysis of an automotive paint chip recovered on a victim can help
investigators narrow the search of potential suspect vehicles. Also the
friction of clothing against plastic parts of a vehicle can produce dis-
tinctive thermoplastic fusion patterns of fabrics that can help elucidate
positioning within and around a car at the moment of impact. The
distribution of glass fragments can help explain the relative locations of
suspects or victims at a scene.
Trace examinations often require a series of detailed microscopical
and instrumental analyses to fully characterize the varied features of
trace materials and to evaluate the significance of the observed prop-
erties within the relevant context. Such analyses range from a physical
comparison of items to determine if there is a physical fit (i.e., fracture
match) to an analysis of the color, construction, and composition of the
materials to determine if differences can warrant an exclusion of a
potential source(s). In the absence of a physical fit or exclusion, a class

⁎
Corresponding author.
E-mail address: ahmehltretter@fbi.gov (A. Mehltretter).

https://doi.org/10.1016/j.forc.2020.100223
Received 5 December 2019; Received in revised form 16 January 2020; Accepted 28 January 2020
Available online 31 January 2020
2468-1709/ Published by Elsevier B.V.
T. Trejos, et al.
association can be reported, indicating the materials could have come
from the same source, or from another source that shares the evaluated
characteristics.
1.1. Informative value of trace evidence
Although the capacity for trace materials to indicate a common
source is not individualizing without a physical fit, it is important to
recognize that trace evidence can offer more than a possible/potential
“source attribution.” Trace materials can be used as exclusionary or
associative evidence, and they can provide valuable information at the
onset of an investigation to corroborate case activities and eyewitness
accounts. The transfer and persistence of trace materials can provide
useful information to reconstruct how a criminal event took place, ex-
pose relevant links, and reveal factors that can lead to the recovery of
additional evidence [3–5].
Materials, such as fabric and glass, can be broken, torn, cut or
otherwise separated during forcible interactions. Likewise, vehicle parts
can be bent during a crash, separating fragments of paint from the
original substrate. As a result, it is possible that the materials trans-
ferred during such an activity can be physically re-aligned with the
original source.
Several experimental studies [6–23] have shown that random phy-
sical fits of materials are rare or improbable. These studies have pro-
vided the foundation for physical fits to be considered the highest level
of association in a trace evidence comparative examination [6–23].
However, there is an ongoing need to develop standardized methods to
evaluate the quality of a physical fit and the uncertainty associated with
such conclusions.
Besides physical fits, an association of mass-produced materials
cannot definitively establish that the items came from a single source.
This is commonly known in forensic science as an association of
evidence with class characteristics. A trace examiner uses several
analytical tools to compare the features of evidentiary items to identify
exclusionary differences or similarities. Since the association to a
possible source can have varying degrees of significance depending on
the characteristics of the samples and their prevalence in the relevant
environment, it is imperative that the examiner assess the significance
for an association and express the limitations to any source attributions
or associations. Efforts are underway among the National Institute of
Standards and Technology Organization of Scientific Area Committees
(NIST-OSAC) to create standard guidelines for interpreting forensic
evidence. This topic has become a primary focus among recent research
studies and projects funded by federal agencies.
Trace evidence has played a critical role in numerous cases, from
the early investigative phases to the corroborative stages and in the
presentation of evidence at trial [3,24–34]. Many of these cases also
illustrate the complexity of the examinations and interpretation of trace
evidential materials. Since the laboratory work is labor-intensive and
the instrumentation can be costly, the examination of trace evidence is
often underutilized. Several authors have proposed models for the use
of trace evidence that can maximize its full potential and emphasize its
value in reconstructions and intelligence investigations [3,4,35–38].
In this paper we present an overarching review of the current state
of trace evidence with emphasis in its scientific foundations. While we
recognize that trace evidence can include many diverse materials such
as fire debris, cosmetics, plastics, pollens, soils, gunshot residue and
lamp filaments, our focus here is on the sub-disciplines that are the
primary emphasis of the NIST-OSAC Materials (Trace) Subcommittee:
hairs, fibers, tape, paint, and glass. We start with a discussion of
training, education, and proficiency testing, followed by a sub-
discipline-specific assessment of their scope and challenges. Material-
specific information regarding the nature, composition, manufacture,
and distribution is critical to informed decisions during a case pre-as-
sessment, the collection, and sampling of fragments at the scene and in
the laboratory, and for the overall interpretation of the evidence. As a
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Forensic Chemistry 18 (2020) 100223
result, these topics will be discussed within each subdiscipline.
Ultimately, the expert’s background knowledge of the manufacture and
distribution of each material of interest is crucial when evaluating
evidence within its relevant population.
Most trace materials can provide information about potential
sources as well as activities that may have led to the contact and ex-
change of traces. Accordingly, the foundations for the transfer of ma-
terials and their persistence is an important focus of this review.
Trace evidence has evolved alongside technological advances in
microscopy, analytical chemistry, and informatics. Historically, forensic
scientists, researchers, and statisticians have contributed to the assess-
ment of the scientific validity of test methods currently in practice. The
process has required several cycles of method development, optimiza-
tion, and validation. Inter-laboratory collaborative tests have aided the
evaluation of meaningful collection sets, testing reproducibility by
multiple scientists, and fine-tuning the scope, capabilities, and limita-
tions of the examinations. Ultimately, the results of inter-laboratory
studies assist the adoption of methods and their application to case-
work.
1.2. Scientific organizations, standardized guidelines, and databases
In the late twentieth century the forensic community recognized the
necessity of establishing consensus guidelines and standard methods for
the advancement of the field and this led to the first scientific working
groups (SWGs) being founded in the United States. In 1988, the
Technical Working Group on DNA Analysis Methods (TWGDAM) was
founded, which later became the Scientific Working Group on DNA
Analysis Methods, (SWGDAM). In 1994, the trace community created
the Technical Working Group for Fibers (TWGFIBE), later known as
Technical Working Group for Material Analysis (TWGMAT), which
became the Scientific Working Group for Materials Analysis
(SWGMAT).
In 2014, most of the federal funding to the SWGs stopped, and NIST-
OSAC was created. The OSAC is currently a collaborative body of more
than 500 forensic science practitioners, researchers, and other experts
who represent all levels of government, academia and industry. For
trace disciplines, the OSAC Materials (Trace) Subcommittee has pri-
marily focused on hair, fiber, tape, paint and glass. Other sub-
committees have been addressing additional trace materials such as fire
debris and explosives, geological materials, and gunshot residue.
Similarly, in 1995 the European Network of Forensic Science
Institutes (ENFSI) was founded with the aim to discuss topics of mutual
interest among forensic laboratories in Europe. The ENFSI now has two
standing committees (Quality and Competence, and Research and
Development) and seventeen working groups, including groups on ex-
plosives, gunshot residues, paint and glass, textiles and hairs, and an-
imal, plant and soil traces. In 2004 the Academia Iberoamericana de
Criminalistica y Estudios Forenses (AICEF) for Latin America, Spain and
Portugal was started, and the founding of the Asian Forensic Sciences
Network (AFSN) occurred in 2008.
Since their foundation, these scientific groups have developed nu-
merous consensus documents that have strived for the harmonization of
best practices and have identified research needs for the advancement
of the profession.
Finally, developing and maintaining collections and databases of
trace materials is a critical but challenging task. These resources are
needed to assess the rarity of observing features of interest in a relevant
population and the ability of the analytical methodology to differentiate
samples. For instance, trace collections and databases are currently
used to assist the classification of materials for investigative purposes.
Researchers use such resources to estimate frequencies of occurrence,
random match probabilities, and other quantitative measures to aid in
the interpretation of the significance of the evidence [3,5,37]. However,
there are difficulties in keeping current with the structural or chemical
changes in materials over time and with increasing global trade.
T. Trejos, et al.
2. Education and training
Education and training in criminalistics include higher education in
science, formal training programs at forensic laboratories, certification
processes, and continuing education activities. Most laboratory posi-
tions require forensic scientists to have a science degree prior to em-
ployment. Degree programs in forensic science have increased on col-
lege campuses, providing general knowledge in criminalistics and
specialized discipline-specific coursework (e.g., trace, chemistry/tox-
icology, DNA). The Forensic Science Education Programs Accreditation
Commission (FEPAC) is the official U.S. accrediting body that assesses
and accredits higher education institutions to ensure quality standards
in undergraduate and graduate forensic science programs.
Forensic laboratories also have their own in-house training pro-
grams for new forensic scientists, which can last for several months to
years. Examiner training typically involves specific procedures and
milestones, including the ability to demonstrate fundamental knowl-
edge for a material of interest, competency in the operation of analy-
tical instruments, methodologies, protocols for sampling, analysis, re-
port writing, and oral testimony. New employees are required to pass
such training programs to become qualified to perform examinations on
casework. Some laboratories follow a modular style of training where
an examiner becomes qualified to examine one material at a time. Then
as time and caseloads permit, they can work cases while training in
other materials and learning new techniques and instrumentation. The
American Board of Criminalistics (ABC), SWGMAT, and more recently
the OSAC for Forensic Science have worked to create guidelines
[39–41] to help standardize training programs for forensic scientists
among different laboratory systems with recommendations on testing
topics and procedures [42].
The ABC offers a voluntary peer-reviewed certification to practi-
tioners as a means to demonstrate professional qualifications in one or
more fields of criminalistics. Recertification involves verification of
competency through participation in the wider forensic community:
attending conferences, continuing education workshops or short
courses, serving on committees, or publishing, as well as successful
completion of annual proficiency tests for practitioners with Fellow
status.
As in any scientific discipline, continuing education and scientific
networking are crucial to maintaining professional competence.
Numerous public and private institutions (e.g., universities, labora-
tories, professional organizations) offer formal specialized workshops
and training to forensic practitioners. Participation in short courses and
workshops at forensic meetings are encouraged as a supplement to
training programs. Additional training outside of one’s own laboratory
system offers opportunities to enhance knowledge and network among
other professionals who handle similar technical challenges. Several
international, national, and regional scientific meetings provide annual
venues to strengthen the forensic discipline through open forums.
Additionally, every three years, Interpol produces a review of the lit-
erature published in the past three years for a variety of forensic dis-
ciplines including glass, paint, fibers and textiles [43].
Finally, the professional interactions within scientific groups serve
as an interdisciplinary forum to improve standards, identify research
and training needs, and offer solutions to advance forensic science
disciplines.
3. Proficiency testing
After completing training, a forensic scientist continues to demon-
strate proficiency in his/her specific disciplines through proficiency
testing. Proficiency tests are used to assess the ability of an examiner to
conduct specific examinations, interpret the data, and communicate the
results. These tests are typically administered annually and are separate
from tests taken during training or the final qualification test ad-
ministered to demonstrate competency to perform forensic
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Forensic Chemistry 18 (2020) 100223
examinations. A proficiency test is meant to replicate the types of ex-
aminations found in casework; however, for practical reasons most are
simplified versions of the typical analytical comparison. An examiner
may have to take multiple proficiency tests if they are qualified in
several disciplines.
Commercial proficiency tests are available for all the major trace
subdisciplines [44,45], and practitioners at a variety of forensic la-
boratories across the United States and the world have received them.
Results have repeatedly demonstrated the accuracy of the analytical
methods and that correct conclusions are reported [44,46]. While
proficiency tests typically reveal high levels of correspondence in re-
ported results among participating laboratories, using these test results
to calculate error rates is problematic. Typically, the parameters of a
case differ from the proficiency test, and properly trained and qualified
examiners are not the only test takers. These test results have also
shown varying approaches to report writing [45,47].
4. Trace evidence interpretation and report writing
With increasing technological advances and worldwide efforts to
validate and standardize examination protocols among different la-
boratories, the focus of trace evidence is also evolving into an estab-
lishment of standardized interpretation methods. The forensic com-
munity is moving away from traditional conclusions such as “the trace
material could have come from the known source” with no further
explanation of what that means [48,49]. The report must be transparent
and informative to the trier of fact. Examiners should provide a report
that documents the methods used, the results and conclusions reached
from those examinations, along with any limitations to their conclu-
sions, and an assessment of the significance of the results. The devel-
oping partnerships of forensic practitioners with other experts in sta-
tistics, law, academia, and industry is vital to the development and
application of functional strategies in this direction.
Forensic analysis is moving toward a two-pronged approach of
professional judgment and statistics to analyze results, interpret data,
and draw conclusions. Descriptive statistics can be useful to analyze
and present data in a meaningful way, such as by showing data patterns
via QC charts, spectral overlay, box-plots, or describing a measure-
ment’s median, range, variance, precision, bias, accuracy, and un-
certainty. Inferential statistics estimate parameters about a population
from a subset of the population to reach conclusions regarding hy-
potheses under consideration [50]. Inferential statistics, including fre-
quentist and Bayesian frameworks, provide a means to address prob-
abilities [50]. Frequentist frameworks estimate probabilities from
repeatable random events (i.e., define probability as the long-term
frequency of occurrence of an event). In contrast, Bayesian frameworks
describe probabilities as degrees of beliefs or logical support [51]. In
Bayesian inference, the prior probabilities or beliefs (i.e., those that
occur before the results/data/information are considered) represent the
assumptions about the unknown parameters. Then, these probabilities
are updated with the data to yield posterior probabilities [50]. In other
words, the Bayesian approach can be used to revise beliefs about un-
certain propositions in the light of new information or evidence [51].
In the frequentist and Bayesian approaches, the interpretation of
evidence can be conducted in multiple steps; in the Bayesian mode, it
can also be conducted in a continuous mode [52–62]. The multi-step
models use comparison criteria to analyze data and then to make binary
decisions of match/non-match. A variety of statistical frequentist
techniques can be applied to answer questions of common source, such
as hypothesis tests, confidence intervals, or range tests [50]. When all
the measured properties of a known and questioned sample are indis-
tinguishable, then the possibility that the compared items originated
from the same source cannot be eliminated. After the comparative stage
(step one), the results are evaluated at a source level (step two) to ex-
plain the significance of the findings (i.e., the rarity of the material’s
characteristics). The final step is the evaluation of these results on an
T. Trejos, et al.
activity level to determine and explain the relevance of the findings
under the given circumstances (i.e., assessment of the evidence con-
sidering competing propositions of alleged activities, and factors such
as transfer mechanisms and persistence) [55,63,64].
While steps two and three are Bayesian in nature, in the frequentist
framework they do not necessarily apply quantitative assessments or
make formal use of statistics to assess mutually exclusive propositions.
The following information can be used in the process to assign a qua-
litative significance to the evidence (i.e., verbal scale of conclusions): a)
peer-reviewed studies that describe the discrimination of features and
methods, b) variations of the measured properties within and between
samples, c) databases, and d) survey studies that evaluate transfer,
persistence, and frequency of occurrence of background materials.
Bayesian statistical methods were first reported in 1963 [65]. Its
application in the interpretation of trace evidence date to the 1970s
[66,67] and continues to be a topic of research [30,57,59,62,68–77]. A
Bayesian framework uses an estimate of a likelihood ratio (LR) to
provide forensic scientists with measurable tools to assess the weight of
evidence. To make estimates, an expert uses searchable databases and
surveys on transfer and persistence as input to the mathematical
models. Advantages of this framework are that it enables a quantitative
assessment for the quality of an association or exclusion, and formally
evaluates the evidence under two mutually exclusive propositions. The
analysis can be conducted sequentially or continuously [75]. There are
several open source packages and accessible programs that can assist
experts in the calculations [56,59,76,77].
Although some forensic laboratories apply Bayesian methods in
their interpretation, this is not yet a common practice. The US judicial
system is an adversarial system where the court is impartial, and juries
decide on a verdict based on competing arguments. This is in contrast to
the inquisitorial systems found in other countries where the court is
actively involved in finding out the facts and can be expected to gain
expertise in statistical methods to help evaluate the evidence presented
before it [78–82]. Legal structure and admissibility of evidence in the
U.S. varies from criminal versus civil courts, and by the court levels
(e.g., county, state, federal). Also, some states use Daubert admissibility
rules (e.g., judge as gatekeeper), while others still use the Frye rules
(e.g., acceptance rule) [83–85].
Accordingly, it is the authors’ opinion that the operation of US
jurisdictions makes the adoption of Bayesian methods more challenging
as compared to some other countries (e.g., Netherlands, New Zealand,
Australia, UK). Some challenges to the estimation of LRs are the de-
termination of the appropriate relevant population in a geographical
area as large as the US, the lack of consistency in LR scale ranges across
disciplines, and that the performance evaluation of LR methods are not
yet standardized. In the trace evidence discipline, some of the sig-
nificant challenges to applying the Bayesian approach are the scarcity
of databases and the existing gap in formal training of examiners in
using these models. Several efforts are being conducted to narrow this
gap and facilitate possible future adoption of these approaches
[56,59,68,69,71,72,77].
The OSAC Materials (Trace) Subcommittee is currently working on
the development and validation of an interpretative approach that
proposes the use of a verbal scale of conclusions to explain the sig-
nificance of the findings. Although the concept is not novel [70], this
group has described material-specific casework examples as a means to
help with the harmonization of the decision process among end-users.
Moreover, the proposed model is suitable for multi-step methods,
continuous methods, and for qualitative or quantitative assessments of
the value of the evidence. As a result, the approach allows the use of
information derived from classic, frequentist, and Bayesian models to
explain the value of findings.
Interpreting the significance of any piece of evidence also rests on a
foundation of knowledge regarding the composition, growth or manu-
facture, distribution, transfer, and persistence of each trace material.
The following sections detail the five main trace materials that are the
4
Forensic Chemistry 18 (2020) 100223
primary focus of the NIST-OSAC Materials (Trace) Subcommittee. The
combination of references cited throughout this paper represent a
comprehensive review of the current state of trace evidence and serve
as background information and a bibliography supporting the inter-
pretation approach being developed by the OSAC Materials (Trace)
Subcommittee.
5. Hair evidence
5.1. Growth, development, and classification
Hair is a biological fiber that grows from the skin of mammals, and
its growth and development continues to be well studied among diverse
biological sciences [86–93]. Humans have thousands of hair follicles
that regulate the cyclical growth of hairs [94], the shape of hair fibers
across the body [95], and hair pigmentation [96,97]. Because each hair
follicle is its own mini-organ driven by complex biological processes,
variation in the positioning and patterning of the internal micro-
structure can be found among individual hair fibers. Hairs emerge from
the follicle as a hardened, keratinized material organized into three
layers: cuticle, cortex, and medulla. Within this layered structure are
melanosomes, the pigmentation organelles for hair; cortical fusi; and
occasionally ovoid bodies; all held together by a cell membrane com-
plex (CMC). The presence and arrangement of the structures that make
up a hair, their patterning, and relative abundance can differ among
mammalian groups and by somatic regions. Hair fibers also vary along
their length in morphological form, size, and in the patterning of the
pigmentation within. Melanosomes are the pigment granules that im-
part color to a hair: eumelanin (brown/black melanin) and pheome-
lanin (yellow/red melanin). It is the size of the various pigment gran-
ules and the amount and distribution of the different melanin types that
account for much of the variation in visible hair color [98–100]. The
pigmentation in hair can vary among the different hairs on a person’s
head, along the length of a hair, and as a person ages [101].
5.1.1. Hair types
Among most mammalian species, guard and underfur hairs are in-
termixed on the body and differentiable by microscopical analysis of
morphology and microstructure. Species and somatic region (i.e., body
area) can often be determined from the shape of a hair and the ar-
rangement of the microstructures within each layer [102–105]. Human
hair forms are separated by regions of the body and are recognizable by
differences in morphology (i.e., its size, cross-sectional shape, degree of
curl) [103,106–109]. Scalp hairs exhibit greater variation among
people than hairs from other body regions so they are more often mi-
croscopically compared in forensic casework [110]. However, pubic
hairs, facial hairs, and other body area hairs can also be compared to
hairs of the same body region. Prior to classifying the somatic origin of
a hair, examiners should have a working knowledge of the varied hair
forms among human populations as well as transitional hairs between
body regions.
5.1.2. Ancestry
Specific hair traits (e.g., cross-sectional shape, degree of curl, pig-
ment density) have been related to population ancestry [111–114],
though intra-individual and intra-group variation can be extensive
[105]. These traits have been used to differentiate hairs among popu-
lation groups [111,114–119]. While there are traits that are commonly
observed among individuals who share a genetic ancestry (i.e., Eur-
opean, sub-Saharan African, Asian/Native American), using a defined
racial classification taxonomy and the historical terminology (i.e.,
Caucasoid, Negroid, Mongoloid) is no longer accepted as valid. Studies
focused on the extent of variation in traits among the hairs on a person’s
scalp and among those of similar genetic ancestry have revealed that
strict taxonomic classifications are not scientifically supported as traits
are more continuous across human populations than historical
T. Trejos, et al.
classifications allowed [120–124]. With increasing levels of admixture
around the world and differing interpretations of racial classifications
associated with cultural and language groups, it is also important to
note that how someone self-identifies may not align with how a hair
examiner relates hair traits to a biogeographic ancestry phenotype. Hair
classification terminology has changed over time because of the
growing recognition that racialized classification schemas are proble-
matic and not always indicative of the extent that ancestry is often
mixed [125–127]. Forensic science is moving toward identifying the
phenotypic traits observed in an evidential hair sample (e.g., degree of
curl, color, cross-sectional shape, density of pigment granules, thickness
of cuticle) and indicating the particular combination of traits is sug-
gestive of coming from someone related to a broad geographic ancestry
group (e.g., European, Asian or Native American, African), yet not
limited to any one population.
5.2. Transfer and persistence studies
Humans shed approximately 100 hairs a day [128] and these hairs
often transfer to our clothing and around our environment. If collected
at a scene or from a piece of evidence, hairs can be informative about
potential contact and activity. How easily hair transfers, persists in its
transferred state, and remains accessible for collection is critical in-
formation to the examiner. Hairs are a specific type of fiber, so much of
the research regarding the transfer and persistence of hair rests on
studies conducted with different fiber types, including wool hairs.
Several studies have evaluated the effect that activity and the types of
recipient garment have on transfer and persistence rates [129–131].
One common source of forensic hair evidence are the hairs collected
from victim’s hands; however, a review of casework found the majority
of those hairs were from the victim themselves, not another person
[132]. In another study [133] an examiner collected hair from his
clothing at varied times during a normal work day for a period of one
month. The examiner found that all the transferred hairs suitable for
microscopical comparison purposes were from close family members
with whom he had personal contact [133]. Another source of hairs on
clothing are hairs that transferred by indirect (or secondary) transfer
from garments that have been washed together [134].
Multiple studies have found that the type and length of contact af-
fect transfer, with indirect transfers being more common than direct
(primary) transfer, and the type of recipient garment greatly influences
the persistence of hairs [129,135,136]. The presence or absence of a
root or whether the hairs have been artificially treated does not appear
to have much influence on how long a hair will remain on a garment
[136]. Instead, the type of textile has been shown to have more of an
effect on the persistence of hairs similar to the reported retention and
loss of fiber data, with woolen garments retaining hairs for a longer
period of time than non-woolen clothing [136,137].
While most human hair research has focused on scalp hair, some
research has focused on the transfer and persistence of pubic hairs
[138]. It was found that, of the 110 heterosexual consensual sexual
interactions studied, pubic hair transfers occurred among 17.3% [138].
Further, transfers were more often from the female to the male, while
cross-transfers (both individuals in a couple transferring pubic hairs to
the other) were rare as it was observed only once in the study [138].
Results for the transfer and persistence of animal hairs have also
been similar to those found for fibers and human hairs
[130,131,139–142].
5.3. Forensic examination protocols
Once hair evidence is received in the laboratory for analysis, a
microscopical examination can provide information on whether evi-
dential material is a hair or other fibrous material, if it is human or non-
human in origin, the body area or possible biogeographic ancestry of
the person it may have originated from, suitability of the root end of a
5
Forensic Chemistry 18 (2020) 100223
hair for nuclear DNA, and if there is any evidence of artificial treatment,
decomposition, or other damage.
A material can be identified as a hair, but hair evidence cannot be
used to identify an individual as the source of the evidence based on a
microscopical comparison. While we tend to think of individuals and
their features as biologically unique, except for identical twins, a co-
incidental association of two hairs that originate from different sources
can occur when similarities are found in the patterning of hair micro-
structure [124,143]. Unlike manufactured materials, which are known
to be mass-produced yet can vary among production lots, the potential
range of variation among the hairs on a person’s scalp and within the
human population is not known. Accordingly, the number of other
potential sources that a hair could be microscopically similar to, and
also be included as possible sources, has not been well established.
For reliable comparison of evidential hairs, the reference samples
must be obtained from the same body region as the hair of interest (e.g.,
head hairs can only be compared to head hairs).
Best practices and guidelines have been developed by practitioners
for forensic examination of hairs. The SWGMAT Human Hair
Examination Guide [144], SWGMAT Hair Training Guide, the ASTM
documents under development [40], and the European Best Practice
Manual for the Microscopic Examination and Comparison of Human
and Animal Hairs [145] are consensus documents which serve as a
model from which laboratories could adapt the methods to their own
training and hair examination protocols.
If exclusionary differences are found between compared samples,
then examinations cease, and it can be concluded that the samples are
dissimilar; and based on the known sample, the source of the reference
sample cannot be included as a possible source of the questioned hair.
However, consideration should be given to how representative the
sample is and whether there could be additional variation not captured
by the reference hairs that could be similar to the evidential hair.
If the characteristics of an evidential hair are not able to be dis-
tinguished microscopically from the characteristics of the known hair
sample, then it can be concluded that the hairs may share a common
source. In this scenario, the number of other people who could share
similar microscopic hair characteristics is unknown. As such, this type
of conclusion is not an identification of one person to the exclusion of
all others. That being said, studies demonstrate that microscopical
analysis can generally differentiate hairs sampled from different people
[124,143,146–150]. An important aspect of forensic reporting includes
a transparent acknowledgment regarding the limitations of an analysis.
Some limitation statements for hair reports may include a statement
that hairs are not a means of individual identification, that an assess-
ment of ancestry is based on hair traits that are only broadly general-
izable to continentally defined populations, and the fact that it is not
known how many other individuals may share similar microscopic
characteristics (and could thus be included as additional potential
sources).
It is recommended that microscopical hair comparisons are followed
by mitochondrial DNA analysis (mtDNA) or, if there is soft tissue pre-
sent at the root end, nuclear DNA (nDNA). Studies have evaluated the
correlation between microscopical conclusions and mtDNA results in
forensic hair analyses with greater than 85% accuracy for microscopical
hair comparisons among hair samples that shared a mitochondrial
profile [124,148,151]. However, the basis of two of these studies
[148,151] were casework samples and results, so the true source of the
hairs and mtDNA profiles were not known for corroboration of accu-
racy. The combination of microscopy followed by mtDNA can increase
the accuracy of potential source inclusion or exclusion for forensic
purposes, and greater significance can be attributed to conclusions re-
levant to a court case; however neither microscopical analysis nor
mtDNA alone can definitively identify a hair to a single person
[124,148,151]. Proteomic research on hair offers a promising comple-
ment to microscopy but has yet to be validated in many forensic la-
boratories [152–155]. Proteomic analysis of hair has limitations similar
T. Trejos, et al.
to mitochondrial DNA in that neither technique can provide an absolute
identification of a single individual; however, in combination with
microscopy these techniques can provide valuable information for
questions as to who the source of a hair may have been and what may
have happened in a case [156].
Laboratories that send hairs directly to nDNA without microscopical
analysis miss important information (e.g., is the hair human?, was the
hair dyed?). Additionally, there is a greater potential to conduct costly
analyses that are not necessary if the unknown hair is microscopically
dissimilar to the known sample of the person from whom it was col-
lected (i.e., victim’s hair found on victim’s body). Ideally, microscopy
should work in tandem with mtDNA, nDNA, or proteomic analyses to
provide the greatest information about evidential hair and its potential
similarity or dissimilarity to a reference sample.
5.4. Discrimination and interlaboratory studies
The ability to discriminate hairs among different peoples by mi-
croscopical analysis has been evaluated through research studies
[124,143,146–150]. These studies demonstrated that comparison mi-
croscopy can discriminate among hair samples, including some iden-
tical twins given a sufficient known hair sample, but the data does not
support identification of hair to a single individual by microscopy
alone.
It should be noted that some historical research [146,147,157] is
often criticized for the statistical analyses employed. This research has
also been misused by some to overemphasize the accuracy of their own
casework, and the statistics have also been misinterpreted by legal
scholars [158,159] to undermine the validity of hair comparisons.
However, this work did show high levels of comparison accuracy
among study samples. The problematic use of and interpretation of
these statistical analyses do not reflect on the accuracy of microscopical
hair analysis. Hair examiners need to be aware of the limitations to the
application of statistical analysis from research: such statistics must not
be applied to casework or be used as an error rate for a case or ex-
aminer, as laboratory methods and procedures may differ from the
research protocol.
5.5. Collections and databases
Reference collections serve as an important resource for comparing
non-human hairs and identifying the features observed microscopically
within the hairs from peoples with known ancestry, disease status,
decomposition phase, and sun exposure. Many laboratories maintain
collections of hair samples from people of known ancestry and hair
samples from different body regions to use in training and as reference
materials. Additional collections have been created by and curated in
natural history museums [160], universities (e.g., Pennsylvania State
University Department of Anthropology), and human remains research
centers (e.g., University of Tennessee at Knoxville, Texas State Uni-
versity). Such collections may be useful in training or research.
While extensive collections are helpful and ideal for training and as
reference material, creating a database of hair features or images is not
a practical tool for forensic identification or for statistical analyses
applicable to forensic casework because of the range of variation that
can be present within hair samples [161]. Features in hair can vary
throughout their growth cycle, and there are a range of nutritional
factors, diseases, or environmental conditions which can influence the
expression of hair traits within a sample [92,162]. While some hair
traits may follow similar patterns (e.g., pigment that aggregates in
streaks, clumps, or patches) among the hairs of one individual, those
hairs can exhibit a wide range of variation in microscopical and mor-
phological appearance (e.g., color, curl, diameter) over time. A data-
base describing the color or density of hair pigment at one point in time
or at a particular length away from the root end would not be useful
long-term, and such a database would be limited in its ability to capture
6
Forensic Chemistry 18 (2020) 100223
a representative sampling of the features of all the hairs from multiple
persons.
Research on hair traits and the variation among samples is ongoing.
While database searches for hair traits or image-based comparison are
not currently applicable to forensic hair examinations, future dis-
coveries may offer new aspects of hair to analyze. Specialists would be
well served by keeping current with the scientific research being un-
dertaken on hair among the textile and cosmetic industries
[95,163–165], and within the fields of dermatology [166–171], an-
thropology [101,172,173], developmental biology [91], and genetics
[87,174–176] as much is still being learned about human hair growth,
its development within the body, and variation among peoples.
6. Fiber evidence
6.1. Composition and manufacture
Fibers serve as the basic unit of textiles and are typically described
as a unit of material, synthetic or natural, whose length is significantly
longer than its width. Natural fibers include plant material from stems,
seeds, or leaves; animal fibers comprised of hair, wool, or silk; and
mineral fibers such as asbestos. Manufactured fibers are created from
regenerated cellulose, synthetic polymers, or inorganic materials such
as glass. Fibers may differ in length (staple or filament), composition
(natural, manufactured, or mineral), the type or amount of additives
incorporated during production (e.g., pigments, dyes, delustrants), their
shape, and treatment or post-production alteration.
Textile manufacturing involves the production of natural and syn-
thetic fibers, spinning those fibers into threads and yarns, creating
fabrics or other textile materials (e.g., weaving, knitting), and the
processes by which fibers and fabrics are colored with dyes or pigments
[177–179]. Man-made fibers (with the exception of glass fibers) are
produced by converting a polymer into a liquid state and then extruding
through a spinneret. The spinneret openings come in a variety of shapes
and produce distinctive cross-sectional characteristics to the fibers (e.g.,
oval, round, trilobal, pentagonal, octagonal). As the polymer exits the
spinneret, it solidifies in air or a solvent before being drawn. The pro-
cess of extruding a fiber through a spinneret is called spinning (wet,
dry, melt, and gel). The drawing process orients the polymer chains
along the fiber axis, increasing its mechanical strength. This process
imparts optical properties useful for identification of fiber-type and for
sample comparisons. Additionally, during fiber manufacture, two or
more polymers can be spun together into a single fiber to provide
special performance features for the end product. Throughout the
production process, fibers take on a wide range of characteristics that
are responsible for the varied fabric types, colors, comfort, and per-
formance features dictated by a global market and constantly changing
fashion trends. Fiber types can be characterized and differentiated by
features such as color, thickness, diameter, cross-sectional shape, di-
chroism, fluorescence, birefringence, sign of elongation, melting point
and chemical composition of its polymers, dyes, pigments, and ad-
ditives.
Natural fibers are produced by breaking down the leaf, stem, or seed
portion of a plant into a fibrous material ready to be spun into yarns
[178,179]. Animal fibers are cleaned to remove debris or other material
(e.g., sericin in silk) and then the fibers are aligned for spinning or
felting [179]. Once usable fiber forms are produced, the fibers can be
spun into yarns differentiated by thickness, number of strands or plies,
and direction of twist. These yarns are then processed into fabric by
knitting, weaving or by creating a non-woven material.
Color can be added at various stages in the manufacturing process.
Pigments can be added to the polymer solution prior to fiber extrusion
or it can be applied to textiles as a surface color. Dyes typically im-
pregnate a fiber, and the stage in the manufacturing process may be
detectable if the whole length of the fiber is not colored. When tightly
twisted yarns or fabrics with a close weave are dyed, portions of
T. Trejos, et al.
individual fibers may not be exposed to the dye bath, thus leaving
portions that are not completely dyed lighter in color. Textiles may also
be overdyed in reaction to market fluctuations (changes in seasonal
styles) or aftermarket processes (recycled fibers). Washing a textile can
alter the color or intensity (hue) as can exposure to UV light. The col-
orfastness and lightfastness of textile materials can impact the number
of potential donor sources of an evidential fiber. Although textiles are
typically produced in batches, changes in the intensity or color of a
material from use and exposure or the application of detergents or
delustrants can provide further discriminating information [180,181].
For more extensive information regarding the composition, manu-
facture, or distribution of fibers refer to the Fiber Organon and the
American Fiber Manufacturers Association website. The American Fiber
Manufacturers Association website is a good source to keep current
with the types of fibers and amounts in production. The Journal of the
Textile Institute and the Textile Research Journal are additional resources
for current and future trends in fiber and colorant technology.
6.2. Transfer and persistence studies
For a fiber to become potential questioned evidence, it must transfer
from one object or place to another and then be collected. How easily it
transfers, persists in its transferred state, and remains accessible for
collection is critical information to the fiber examiner. Fibers can be
shed from yarns and textiles through a range of activities including
abrasion, washing, and stretching/straining a textile material. Physical
damage to a textile material can expose yarn ends and reduce yarn
cohesion or twist and allow individual staple fibers to be lost [182].
Normal wear and activity patterns lead to fibers being shed and
transferred to other people or objects through contact.
An extensive body of literature has been written on the transfer of
fibers as well as how long those fibers may persist on a given person or
object [130,131,183–198]. Specific research has been conducted on the
frequency of different fiber types to transfer [199,200] and the activ-
ities and types of recipient garments that could affect fiber retention
[130,186,189,200,201], as well as varied environmental conditions or
exposure [191,202] and secondary transfer [196–198]. After transfer-
ring, fibers can be lost and/or redistributed on a garment through the
actions of gravity, the activity of the person wearing the garment
[183,184,186,190] or through transportation of the evidence [203].
Features of the recipient garment (tightness of weave, fiber type)
greatly influence the rate of fiber persistence [130]. Transferred fibers
are typically lost within the first hour or two [131] under normal ac-
tivity patterns. Washing and exposure to outdoor elements can impact
the persistence of transferred fibers [183,190,191,194,195,202,204].
6.3. Forensic examination protocols
Once fiber evidence is received in the laboratory for analysis, the
physical, optical, and chemical features of fibers are assessed and
compared to determine whether fiber properties differ or correspond.
Best practices and guidelines have been developed among US and
European practitioners for forensic examination of fibers and for the
specific techniques used during analyses [205–209].
When examining damaged fabric, an examiner evaluates the po-
tential for a physical fit to be made between torn edges. A physical fit of
two fabric pieces is considered the highest level of source attribution in
textile examinations [210,211]. When a physical fit conclusion is made,
it is the examiner’s opinion that the possibility is negligible that the
compared textile materials originated from different damaged sources.
When a physical fit is not found or possible, the examiner evaluates
the physical features of the material such as color and construction,
optical properties of the fibers such as fluorescence, and chemical
properties such as fiber composition or type. The order of examinations
is typically done from least destructive to most destructive, but the
extent of discrimination provided by each analysis is also considered.
7
Forensic Chemistry 18 (2020) 100223
An examination typically starts with light microscopy and comparing
physical characteristics with a comparison microscope. Microscopical
analysis is non-destructive and provides discrimination among fibers
based on differences in color, size, shape, and other microscopic char-
acteristics such as the presence of delustrants [177,212–214]. Optical
properties (e.g., refractive index, birefringence, dichroism) are eval-
uated with polarized light microscopy (PLM) and fluorescence light
microscopy, and spectra of colored fibers are compared with UV/Vis
microspectrophotometry (MSP). Fourier transform infrared micro-
spectroscopy (FTIR) is typically used to confirm manufactured fiber
type determined by PLM and to determine fiber sub-type. While this
technique can be highly discriminating as to fiber type, it does not have
the discriminating power to differentiate fibers that have the same
major chemical composition but are different in size, color, and shape.
The majority of fibers can be discriminated by comparison microscopy,
PLM, and MSP, so the additional resolving power FTIR may bring to a
forensic fiber analysis to further discriminate samples is typically low.
Additionally, FTIR is semi-destructive to fiber morphology so is typi-
cally used later in a forensic fiber protocol.
If any exclusionary differences between compared samples are
found at any point, then examinations can cease, and it can be con-
cluded that the samples do not share a common source. If the micro-
scopic characteristics, optical properties, and chemical composition of
an evidential fiber are not able to be differentiated from the known
fiber sample at each stage of the fiber examination protocol, then it can
be concluded that the fibers could share a common source. This con-
clusion does not mean that an evidential fiber came from the same
textile material as the known sample, since fibers and textile materials
are often produced in large quantities/batches/lots.
6.4. Discrimination and interlaboratory studies
Fibers can be microscopically discriminated by color, shape, size,
and presence of delustrants, even under low magnification [215–217].
A variety of dye combinations may lead to similar macroscopic colors
on fibers and textiles; however, MSP and thin layer chromatography
(TLC) can further identify and characterize the dye or pigment com-
ponents [217–219]. Color is arguably one of the most discriminating
characteristics used in fiber examinations; however, the uptake of col-
orants is more uniform in manufactured fibers than in natural fibers
where the structure and appearance are typically more variable. Ac-
cordingly, the use of MSP to compare spectra provides a method by
which color measurements are standardized, repeatable, and the data
can be quantitatively analyzed [220,221]. This method serves as a
complement to visual assessments of fiber color as well as a verification
of an examiner’s ability to perceive slight color differences.
Research focused on several common colors and fiber types found
high discriminating power when using a combination of light micro-
scopy, fluorescence microscopy, and MSP [222,223]. Depending on the
dye type and fiber color, discriminating power ranged from 89% for
black fibers dyed with direct dyes, 93% for black fibers dyed with re-
active dyes, 93% for orange cotton fibers, and 99.8% for green cotton
fibers [215,223]. Non-denim blue cotton fibers were found to have a
discriminating power of 96% when compared visually with microscopy
and MSP [217].
Studies of fiber populations found in casework or public locations
[219,224,225] and the extent that comparably colored fibers can be
discriminated [215,222,223,226] have done much to inform the for-
ensic community about the discrimination power of forensic fiber
analyses and their limitations, particularly in relation to common fiber
types and colors [215,222,223,226]. Some colors and fiber types (e.g.,
blue denim cotton) are ubiquitous in clothing material, irrespective of
style and seasonal changes so their significance as evidence has been
limited.
When interpreting the significance of fiber evidence, consideration
should be given to how common the fiber is within the general
T. Trejos, et al.
environment by assessing general fiber populations [227–232]. Fur-
thermore, some locations or regions may have a greater affinity for
particular colors (e.g., colors of sports teams) than other areas, or have
different prevalence of fiber types (e.g., cotton vs wool) based on
weather conditions and climate (e.g., hot/humid, cold/dry). Fiber po-
pulation studies [199,224,227–234], color block studies
[215,217,223,226,235–238], and target fiber studies
[198,215,219,225,236,239–241] address the frequency of occurrence
of similar color and fiber type, and these studies should be considered
when interpreting the significance of fiber evidence.
Interlaboratory studies typically reveal high levels of correspon-
dence among laboratories; however, as with proficiency tests, any sta-
tistical values calculated from the results found in these studies should
not be used to bolster or undermine the validity of results obtained in
criminal casework as the parameters of a case typically differ from re-
search.
Research being undertaken by textile conservationists, archaeologists,
polymer chemists and for industrial applications has expanded the
knowledge regarding how long fibers may be preserved in different
environments [242–244], methods to potentially analyze aged or
weathered fibers [181,245,246], sample analysis with the least destructive
methods possible [247], and new fibers used in various industrial and
medical applications [248–253].
6.5. Collections and databases
Many forensic laboratories have reference collections maintained
within laboratory systems comprised of various fiber types from vali-
dated source materials. These collections are used for training and re-
search purposes. Spectral libraries are often constructed from these
collections or commercially available libraries are purchased to aid in
the identification of fiber composition by FTIR. It is practically im-
possible to create a population database representative of all the fiber
types and dye combinations that are commercially available at any one
point in time, but attempts have been made to use smaller laboratory
reference collections and databases created from fibers found in case-
work for relative abundance analyses among varied fiber populations
[184,199,215,223,224,235,254,255]. Such collections have primarily
been used for research purposes, due to the constantly changing textile
market (e.g., style, color, season), differences in the durability of dif-
ferent fibers types, and colorfastness of the dyes used. When compar-
isons to any commercial or in-house databases are applied to casework
and statistical analyses are conducted, the limitations and uncertainty
must be explicitly stated. Nonetheless, these collections serve as useful
reference material for identification, classification, and comparison of
fibers and can be an important resource in the examination and training
processes.
7. Tape evidence
7.1. Composition and manufacture
Tape is defined at a general level as a narrow flexible strip or band,
but for purposes of trace evidence analysis, the term “tape” refers to
pressure-sensitive (PS) tape. PS tape can be defined as a film to which a
PS adhesive is attached, which allows the film to adhere to a surface
following the application of light (finger)-pressure. At a minimum, PS
tape consists of a film/backing and adhesive layer but may include
fabric (e.g., duct tape) or glass (e.g., strapping tape) reinforcement. This
review focuses primarily on duct tape, electrical tape, and packaging
tape, since they are the most commonly encountered types of tape in
U.S. forensic laboratories [256].
The tape industry utilizes hundreds of raw materials in manu-
facturing. Tape manufacturers may either manufacture their own
components or purchase them and then mix the raw materials in dif-
ferent recipes (ingredients and relative amounts). Tapes are generally
8
Forensic Chemistry 18 (2020) 100223
produced as jumbo rolls that are subsequently cut down by the manu-
facturers or distributors into smaller rolls for purchase.
For extensive information regarding history, composition, or man-
ufacture, refer to Benedek [257], Johnston [258], or Satas [259]. The
Pressure Sensitive Tape Council’s Products Directory and the Adhesives
and Sealants Industry newsletter are periodicals that are good resources
for current and future trends in tape and adhesives technology.
7.1.1. Duct tapes
There are multiple ways to produce duct tape depending on
manufacturer preferences, and these different production methods/
processes result in the variability observed among manufacturers and
products. Typically, backings are made of polyethylene with colorants
added (e.g., aluminum for silver-colored tapes). The fabric
reinforcement is made of cotton, polyester, a cotton-polyester blend,
or less commonly nylon, but these fibers/yarns can be constructed in a
variety of ways. Adhesives are most commonly made of natural
rubber plus a tackifying resin and inorganic fillers (e.g., titanium
dioxide, calcium carbonate), though other compositions can be
expected.
7.1.2. Electrical tapes
Electrical tapes consist of a plasticized PVC backing and usually a
synthetic adhesive (e.g., styrene-butadiene rubber). There are fewer
physical features present for evaluation of electrical tapes (i.e., no re-
inforcement layer) than duct tapes but there is still a wide range of
chemicals that can be used, which leads to the variability observed
among the different manufacturers and products.
7.1.3. Packaging tape
Packaging tapes consist of a polypropylene backing and usually a
synthetic adhesive (e.g., acrylic). There are fewer physical features
present for evaluation in packaging tapes than there are for duct tapes,
but there is still a range of optical and chemical analyses that can dif-
ferentiate some manufacturers and products.
7.1.4. Other
Other tapes include strapping tape, Gaffer’s tape, friction tape, of-
fice tape, and masking tape. Each of these has their own range of
chemicals used in their manufacture based on their intended end use.
7.2. Transfer and persistence studies
Tape evidence is often purposefully used in the preparation for or
commission of a crime, rather than passively transferred. Therefore,
transfer studies like those for other trace materials are less relevant, and
no studies regarding transfer and persistence of tape itself were found
within the literature review. On the other hand, the adhesive of the tape
usually captures other trace materials on its surface, making it a
common substrate for other trace evidence. Persistence would primarily
involve evaluating how long the tapes would stick to surfaces via the
adhesive under a variety of conditions (e.g., under physical stress such
as stretching, environmental conditions such as heat and water).
Manufacturers extensively study properties such as adhesion, tack, and
shear resistance in developing their products [258] but this has not
been studied much in relation to forensic evidence.
7.3. Forensic examination protocols
Once tape evidence is received in the laboratory for analysis, the
physical and chemical features are analyzed and compared to de-
termine whether or not the properties correspond. First, the tape is
examined visually and microscopically, then with a series of instru-
mental examinations in order to evaluate and compare the chemical
properties. A textbook chapter has been written that extensively covers
the forensic analysis of tape [260].
T. Trejos, et al.
Best practices and guidelines have been developed by practitioners
for forensic examination of tapes and for the specific techniques used
during analyses [261,262]. SWGMAT’s Guideline for Forensic Ex-
amination of Pressure Sensitive Tapes provides a basic overview of
examinations, with the additional ASTM and SWGMAT guidelines
providing more detailed information about specific techniques.
SWGMAT also created an extensive bibliography of relevant
tape publications up until May 2012 that can currently be found
on the American Society of Trace Evidence Examiners website
(http://asteetrace.org) [263] under the Resources tab.
As one of the first steps in the analysis, the examiner evaluates the
potential of a physical fit between the ends of different pieces of tape
relative to each other to determine whether they can possibly be rea-
ligned to form one continuous piece. A physical fit of two tapes is
considered the highest level of source attribution in tape examinations
[264]. When a Physical Fit conclusion is made it is the examiner’s
opinion that the possibility is negligible that the compared tapes ori-
ginate from different torn/cut tape sources. Several studies have been
published for validation of such examinations [8,20,265].
When a physical fit is not observed, the examiner evaluates the
physical features such as color and width and chemical properties such
as the adhesive composition. Examinations are typically ordered from
least destructive to most destructive, but also considers the extent of
discrimination provided by each analytical method. For instance, phy-
sical examinations yield high discrimination power [266–269] and are
generally simple to perform, so they are conducted first. On the other
hand, pyrolysis-gas chromatography would most likely be the final
analytical step because the technique is destructive, though only mi-
crogram amounts of materials are needed, and the technique is more
time-consuming compared to others [270].
If any exclusionary differences between compared samples are
found at any point, then examinations cease, and it can be concluded
that the samples do not share a common source. If the characteristics
and properties of an evidential tape are not able to be distinguished
from the properties of the comparison sample at every stage of the tape
examination protocol, then it can be concluded that the tapes may share
a common source. A conclusion that tapes could have a common source
does not necessarily mean the questioned tape came from the known
roll, but either it or one like it that shares the same properties eval-
uated. Rolls are often made in batches, so rolls of the same product are
expected to be indistinguishable, but discrimination studies demon-
strate that different tape products can be readily differentiated
[266,268–270].
7.4. Discrimination and interlaboratory studies
Tapes can be microscopically discriminated by color, construction,
physical measurements, and other properties [266–269]. Physical
characteristics have been demonstrated to be highly discriminating for
duct tapes (e.g., 99.6% for a set of 82 different sources, 3321 com-
parison pairs) due to the complexity of duct tape construction [269].
Instrumental techniques, such as FTIR, scanning electron microscopy/
energy dispersive spectroscopy (SEM/EDS), and pyrolysis–gas chro-
matography/mass spectrometry (Py-GC/MS), which are generally
standard in forensic science laboratories, can further differentiate tapes
based on their chemical compositions [268–274].
For tapes with fewer physical features to evaluate (e.g., electrical
tape, packaging tape), these chemical examinations are more critical for
discrimination. For electrical tapes in particular, different discrimina-
tion powers were obtained for the various studies depending on the
sample set and the techniques used, but FTIR, SEM/EDS, and Py-GC/
MS, all have been demonstrated to be useful for discriminating different
tape products. For instance [268], Py-GC/MS alone was able to dis-
criminate 83% of adhesives and 81% of backings in a set of electrical
tapes known to contain rolls from different products and duplicate rolls
of the same product; in combination with other techniques, 96%
9
Forensic Chemistry 18 (2020) 100223
discrimination was able to be achieved on the same sample set. Tapes
have been found to typically be the same product/manufacturer when
they are unable to be distinguished following a comprehensive com-
parison as demonstrated by the combination of these studies
[266–274].
Another set of studies have evaluated within-roll variation and
found that tapes are consistent along their lengths, but differences can
sometimes be found between different rolls of the same product
[275–278].
Additional technologies such as X-ray fluorescence spectroscopy
(XRF) [279], isotope ratio mass spectrometry (IRMS) [280,281], sector
field inductively coupled plasma-mass spectrometry (SF-ICP-MS) [282],
laser ablation-ICP-MS (LA-ICP-MS) [283–285], and laser induced
breakdown spectroscopy (LIBS) [283,284] are receiving increased at-
tention in forensic research laboratories around the world, with results
that are promising and comparable to or even better than conventional
methods [279–283,285].
7.5. Collections and databases
There are few duct tape manufacturers in North America and even
less in other parts of the world. For these reasons as well as the wide
range of characteristics available for comparison between different duct
tapes, this tape is amenable to sourcing (i.e., determination of manu-
facturer/product). The largest known duct tape collection in North
America is the FBI’s National Forensic Tape File (NFTF), which has
approximately 700 duct tape samples and is used for manufacturer/
product searches. Other collections exist but on a smaller scale and are
generally not used for source determinations. The accompanying da-
tabase is housed in an Access®-based software program, which allows
for text- and numerical-based searches in conjunction with energy
dispersive spectroscopy spectral searches to determine a list of potential
manufacturer/product candidates. Manufacturing representatives are
then contacted for corroboration and distribution information. The FBI
has published a casework example of the use of the NFTF and other
resources [286]. Other types of tape are not amenable to sourcing due
to the range of manufacturers and countries of origin, but collections
and databases can still be created and used to assess the range of pos-
sible physical features and chemical properties and evaluate the dis-
crimination power and relative merits of the methodology.
8. Paint evidence
8.1. Composition and manufacture
Paint is defined as a surface coating designed to protect or decorate
a surface. It can also be stated as a suspension of a pigment in a vehicle.
The term “vehicle” in this context is used by the paint industry to de-
scribe the liquid portion of the paint comprised of the solvent and the
binder. The solvent is the volatile portion of the vehicle and is used to
aid in the production, consistency, and application, after which the
solvent is lost from the paint. The binder is the fraction of the paint
vehicle that provides adhesion both within the paint film and between
the paint and the substrate; it is often called a resin and is the non-
volatile portion of the paint vehicle. A pigment, a finely powdered solid
dispersed in the coating binder, may be inorganic, such as titanium
dioxide, or organic, such as quinacridone. Pigments may impart color
and opacity or, in the case of extender pigments, may be used to add
bulk to the formulation at a reduced cost or to change physical prop-
erties such as strength and weatherability. Additives may also be in-
cluded in paints such as driers, plasticizers, anti-skinning and anti-set-
tling agents, and preservatives.
The paint industry utilizes hundreds of raw materials in manu-
facturing. Most paint manufacturers do not manufacture their own re-
sins and pigments but instead purchase them and then mix the raw
materials in different recipes (ingredients and relative amounts).
T. Trejos, et al.
For extensive information regarding composition, manufacture, or
distribution, refer to Marrion 2004, Streitberger 2008, or Wicks 2007
[287–289]. Automotive News, Paints and Coatings Industry, and Coatings
World are periodicals that are good resources for current and future
trends in paints and coatings technology.
8.1.1. Automotive paints
One of the most common types of paint submitted to forensic la-
boratories is automotive. Several steps are involved in original equip-
ment manufacturing (OEM) automobile painting and are most fre-
quently performed in the following order: metal pretreatment,
electrodeposition, primer-surfacer application, and finish coat applica-
tion(s), each of which provides a layer of paint [288]. Different plants
use different paints for their layer systems. Vehicles may require repair,
either at the factory or aftermarket, which can further affect the layer
structure of the paint system. Typically, the more complex the layer
system, the more unusual it is.
A wide variety of polymers can be used as paint binders and cross-
linking agents, including, but not limited to, acrylic, alkyd, epoxy,
melamine, polyester, styrene, urea, and urethane [290].
8.1.2. Architectural paints
Architectural paint, frequently called household paint and sold in
many retail stores, is also frequently submitted for analysis. Often these
paints are base white and can be tinted in-store to produce a wide range
of colors. Architectural paint can consist of numerous layer structures,
depending on the number of times the substrate has been painted. The
lifetime of a structure is generally much longer than that of an auto-
mobile, and the paint is rarely removed before subsequent layers are
added, resulting in the potential for a sample to have many layers made
up of various colors and chemistries.
The resins used for architectural paints can be classified as either
waterborne latex systems or organic solvent-borne systems. Latex sys-
tems include acrylics, polyvinyl acetate (PVA) – acrylics, PVA-ethy-
lenes, and styrene-butadienes. Organic solvent-based resin systems may
consist of acrylic enamels, acrylic lacquers, alkyds, nitrocellulose lac-
quers, or urethane enamels [290].
8.1.3. Other
Other paints include spray, bicycle, motorcycle, maintenance (e.g.,
tools, bridges), marine, and aircraft paints. Each of these has their own
range of chemicals used in their manufacture based on their intended
end use and expected environmental conditions [290].
8.2. Transfer and persistence studies
For paint to become potential questioned evidence, it must transfer
from one object or place to another and then be collected. How easily it
transfers, persists in its transferred state, and remains accessible for
collection is critical information to the paint examiner. Spray paints and
household/architectural paints can transfer wet or dry to clothing and
other objects, and vehicle paint can transfer to other vehicles, individuals
and their clothing, and other objects. Studies were undertaken to de-
termine the background levels of paint flakes on clothing items unrelated
to a crime [291] and related to a crime [292]. Both studies determined
that paint can be found, single-layered paints are more common than
multi-layered paints, and the number of chips found is inversely related
to the number of layers observed. Careful examination of clothing can
establish a person’s exposure to aerosolized spray paint, though not ne-
cessarily establish him/her as the sprayer [293]. The most common areas
to observe aerosolized paint on the sprayer were that person’s lower
torso, wrists, and knees [294]. Very fine droplets do not persist as long as
larger wet paint transfer as they can be readily washed away [293]. Some
studies have reported the efficiency of spray paint sampling methods and
the influence of substrate and weathering effects [295–298]. Regarding
transfer of tool (e.g., crowbar) and household paints, chemical
10
Forensic Chemistry 18 (2020) 100223
composition and age influenced how much paint transferred, and cross-
transfer and secondary transfer were both observed [299]. Cross- or two-
way-transfers of automotive paint are considered by examiners as
stronger evidence than one-way transfers [300,301]. It is expected that
this is because multiple types of paint would have transferred, and two
different associations would be reported, compounding the circum-
stantial evidence supporting contact between the two objects.
8.3. Forensic examination protocols
Once paint evidence is received in the laboratory for analysis, the
physical and chemical features of paints are analyzed and compared to
determine whether or not the properties correspond. This is done by
first examining the evidence visually and microscopically, then with a
series of instruments in order to evaluate and compare the chemical
properties. Several publications have been written that extensively
cover the forensic analysis of paint [290,302–304].
Best practices and guidelines have been developed by practitioners
for forensic examination of paints and for the specific techniques used
during analyses [41,305–308]. ASTM E1610 provides a general over-
view of paint examinations, details the strengths and limitations of each
method, and describes the fundamentals of an appropriate analytical
scheme.
SWGMAT created an extensive bibliography that covers the state of
paint analysis up to 2012 and includes summaries of many publications,
which can currently be found on the American Society of Trace
Evidence Examiners website [263] under the Resources tab. Every three
years, Interpol issues a review article covering paint analysis; the most
recent ones cover 2013–2016 and 2016–2019 [43].
As one of the first steps in the analysis, the examiner evaluates the
potential of a physical fit between the known paint and a questioned
chip. A physical fit of two paint chips is considered the highest level of
source attribution in paint examinations [308]. When a Physical Fit
conclusion is made, it is the examiner’s opinion that the possibility is
negligible that the compared paint chips originate from different da-
maged paint sources.
When a physical fit is not found, the examiner evaluates the physical
features such as color and layer structure and chemical properties such
as the binder composition. The order of examinations is typically done
from least destructive to most destructive, but also considering the
extent of discrimination provided by each analysis. For instance, phy-
sical examinations yield high discrimination power [309–311] and are
generally simple to perform, so they are conducted first. On the other
hand, pyrolysis-gas chromatography may be the final analytical step
because the technique is destructive, though only requires microgram
amounts of materials [304], and the technique is more time-consuming
compared to others.
As with other trace materials, if any exclusionary differences between
compared samples are found at any point, then examinations cease, and
it can be concluded that the samples do not share a common source. If
the characteristics and properties of an evidential paint could not be
distinguished from the properties of the comparison sample at every
stage of the paint examination protocol, then it can be concluded that the
paints could share a common source. A conclusion that paints could have
a common source does not necessarily mean the questioned paint came
from the known, but either it or one like it in all the same properties
evaluated. Paints are often made in batches, so containers of the same
color and product, and vehicles of the same color from the same plant are
expected to be indistinguishable, but discrimination studies demonstrate
that different paint products can be readily differentiated.
8.4. Discrimination and interlaboratory studies
For automotive paints, studies starting in the mid-1970s until the
present day have repeatedly demonstrated that a comprehensive ana-
lysis is capable of discrimination powers well above 99% for
T. Trejos, et al.
automotive paint, and the samples that were not differentiated were
found to have originated from the same make and model in approxi-
mately the same time frame (i.e., same model year or within a couple
model years) [312–316]. It is expected that the samples from each of
these indistinguishable pairs originated from the same automobile
factory. These studies typically consisted of samples collected at junk
yards, auction yards, and other damaged vehicle sources and ranged in
samples set sizes from 200 to 500.
It was expected that factory repairs would be less common than the
standard OEM systems observed and that the addition(s) of factory
repaint layers would increase the significance of an association. A study
[317] was conducted to test this expectation and evaluated approxi-
mately 1000 OEM paint samples. It determined that while most systems
are standard systems (~92%), one additional clearcoat/basecoat (CC/
BC) combination occurred in ~6% of samples, two CC/BC system re-
pairs occurred in ~1%, and three clearcoat/basecoat system repairs
(four total CC/BC sets) occurred in ~0.28% of samples. The total per-
centage of samples with factory repairs was in alignment with rates
quoted by the automotive industry and substantiates the expectation
that the addition of each subsequent factory repair layer system in-
creases the significance of an association.
It is further expected that the presence of aftermarket paint layers
can increase the significance of an association. A vehicle owner having
his/her automobile repainted would have a selection of different shops
they could use, and different shops have different paint supplies and
combinations they use in their repaint materials. A factory, on the other
hand, typically uses the same paints in its repaints as it does in its
original paint system. Depending on the reason for an aftermarket re-
paint (e.g., change of color, damage from a crash), the original layers
could be completely or partially removed before the vehicle is re-
painted, or the repaint layers could be put directly on top of the existing
paint layers. In general, the more layers present for evaluation, the
more significant the findings can be.
Architectural paint studies, most of which included approximately
1000 or more samples, have similarly demonstrated that a compre-
hensive analysis is capable of discrimination power well over 99%
[297,311,318,319]. Even single-layered white architectural paints,
considered “worst-case scenario” comparisons, resulted in 99.35%
discrimination among fifty different sources [320].
A “worst-case scenario” evaluation of black spray paints also found
a discrimination of 99.4% [321], which corroborate the conclusions
reached in other similar studies [322–324]. The aforementioned paint
discrimination studies also discuss the discrimination power at various
stages of the analytical scheme, providing the examiner basis for
making choices of analytical methods and offering information to assess
the significance of comparative analysis.
Spray paint is expected to be more prone to heterogeneity concerns
than other types of paint encountered in forensic laboratories due to the
need to shake the can prior to application. This is exacerbated by a
likely reduced need of the painter in criminal activities to have a
homogeneous application since the paint is typically being used to di-
rectly commit a criminal act (graffiti) as opposed to having been ap-
plied for aesthetic purposes in non-criminal activities. In order to ad-
dress these issues, several studies have been conducted to evaluate the
effect of shaking on chemical analysis and comparison of spray paints
[325,326]. Results confirm that there is variation in color and spectral
data collected for spray paints shaken to various extents. Such knowl-
edge is helpful in assessing the significance of spectral variations be-
tween questioned and known samples being compared.
While shaking has an effect on the chemical composition of spray
paints, variations may also occur due to batch differences. In order to
evaluate batch-to-batch variation in spray paints, an inter-laboratory
study involving eleven laboratories was conducted [327]. Results from
this study indicate that for colored samples, techniques aimed at color/
pigment characterization performed better than those aimed at binder
or elemental composition; for white samples, those aimed at binder
11
Forensic Chemistry 18 (2020) 100223
composition were most effective; and for black samples, Py-GC/MS was
the best technique.
Additional technologies such as LA-ICP-MS [328–330] and Raman
spectroscopy [331–340] have also received attention in forensic re-
search and art laboratories around the world, with results that are
promising and comparable to conventional methods. Raman spectro-
scopy has shown complementary information to FTIR, particularly in
the identification of pigments. However, LA-ICP-MS has not been
widely adopted in forensic laboratories due to its cost, as well as the
high discrimination rates possible with the more conventional methods.
Regarding the accuracy of analytical methods and consistency
among laboratories, a collaborative study of black automotive paints
demonstrated inter-laboratory reproducibility of spectra collected on a
variety of FTIR instruments and accessories [341]. Some minor varia-
tions were observed, with thin layers presenting expected difficulties.
Additionally, an inter-laboratory study organized by the Royal Cana-
dian Mounted Police (RCMP) in Canada further demonstrated that
different examiners and different laboratories arrive at correct conclu-
sions when provided mock case samples [342].
8.5. Collections and databases
Paint collections and databases exist that are used for make/model/
year searches or as an aid for determining frequency of occurrence of a
particular sample. The most widely used is the Paint Data Query (PDQ),
which was developed by and continues to be maintained by the RCMP
[343,344].
A series of studies has been recently conducted to enhance PDQ
library search algorithms to ATR-FTIR spectra as well as prefilters to
identify potential vehicle make and models [345–347].
PDQ is a collection of automotive paints with an accompanying
database of information for over 22,000 multi-layered paint samples
(more than 90,000 individual layers) (Version 1.0.0.60 – 2018) from
foreign and domestic OEM automobiles marketed predominantly in
North America since about the 1970s. This compilation of information
is based on layer structure and the chemistry of each layer. Text-based
searches specifying compositions and spectral searches can both be
performed after determining the layer structure and chemistry of the
individual layers from an evidentiary paint sample.
In addition to providing potential make/model/year/plant in-
formation from a questioned sample, the information in the database
allows for statistical information to be generated in order to aid in a
significance assessment and to track trends and changes in formula-
tions. PDQ is not designed to be a population database representative of
all the vehicles on the road, so it is not a population database, but in-
stead attempts to have a sample of every type. One of the collaborative
validation studies mentioned previously [342] demonstrated that most
forensic participants were able to correctly determine the manu-
facturing plant and model year range of small automotive paint samples
using PDQ, and the use of PDQ has been documented in casework and
at trial [345–350]. PDQ has also been used to evaluate the prevalence
of OEM factory repairs [317].
Three more collections include the refinish color books available
from the various paint manufacturers, such as Axalta, BASF, and PPG
Industries; the National Automotive Paint File (NAPF), which is main-
tained and used exclusively by the FBI Laboratory; and the European
Collection of Automotive Paints (EUCAP), maintained by the
Bundeskriminalamt in Germany. The former two compile the colors
used on vehicles marketed in North America, and the latter is comprised
mostly of paint from vehicles found in Europe.
Any forensic laboratory can contact the manufacturers and receive
copies of the repaint pages for its own collection; a case example of its
use in conjunction with PDQ is available [25]. Automobile manu-
factures and their paint suppliers contribute to the FBI’s NAPF by
submitting color standards. This collection contains paint colors of
automobiles available in North America on domestic cars from 1934
T. Trejos, et al.
and foreign cars from 1955. Some manufacturers submit full paint
system panels for inclusion in NAPF, which the FBI forwards to the
RCMP for inclusion in PDQ. Similarly, there is cooperation among the
RCMP, FBI, and BKA in sharing samples among collections. Some for-
ensic laboratories also maintain collections and databases re-
presentative of their casework.
Other types of paint are not amenable to sourcing due to the range
of manufacturers and distribution channels, but collections and data-
bases can still be used to assess the range of possible physical features
and chemical properties, and to evaluate the discrimination power and
relative merits of the methodology, as demonstrated in many of the
previously referenced works.
9. Glass evidence
9.1. Composition and manufacture
Glass is a material that is made up of a complex mixture of atoms
that combine asymmetrically without crystallization [351]. The relative
amounts of its constituents determine the properties of the final pro-
duct, including hardness, resistance, transparency, viscosity, density,
refractive index (RI), and color.
There is extensive information regarding the composition, manu-
facture, and distribution of glass available within the scientific litera-
ture [53,302,352–355]. The online periodical Glass Magazine
(www.glassmagazine.com) is a good resource from the National Glass
Association (www.glass.org) for current and future trends in glass pro-
duction. Some manufacturers such as Pilkington and the Nippon Sheet
Glass Company (NSG group) publish annual reports describing statistics
on market demands, market routes by glass type, geographical location
and annual production of glass manufacturing plants [356,357].
Glass manufacturing begins with the mixing of raw materials and
homogenizing them at high temperatures (~1500 °C). The major con-
stituent of glass is silicon dioxide, which is responsible for building the
network of the glass structure. Other components (e.g., sodium, cal-
cium, and magnesium) make it more difficult for atoms to rearrange
and crystallize, and also reduce the melting point and viscosity of silica.
Many other elements are added to the formulation of glass, such as
secondary formers (e.g., aluminum, zirconium, titanium), decolorants
that improve transparency (e.g., iron, chromium, selenium, manga-
nese), refining agents that prevent the retention of gas bubbles in the
glass (e.g., calcium), and stabilizers that provide chemical resistance
(e.g., calcium, magnesium, lead, strontium, barium, zinc). Broken glass
ready for recycling, also known as cullet, is used in manufacturing
plants to decrease production costs [354].
Glass manufacturing plants are often established near natural re-
sources of the primary raw materials, as it represents a cost-effective
strategy. These raw materials, such as quartz (silicon dioxide), lime-
stone (calcium oxide and magnesium oxide), and trona (sodium car-
bonate), not only contribute to the major glass composition but also to
trace contaminants that are an important elemental signature of the
end-product. Most of these trace elements are a result of the geological
history of the mining sites, and therefore, it is expected that the ele-
mental composition of glass from factories located in different geo-
graphical sites will vary widely [75,352,354,355,358–360].
Over 500 glass production plants operating throughout North
America, Asia, Europe, South America and the Middle East serve the
global market for glass [361,362]. Innovation has produced significant
changes in glass formulations to make it stronger, lighter-weight, and
more energy efficient. Shatter-resistant screens in mobile phones, ultra-
clear glass for self-driving vehicles, self-cleaning glass in buildings, and
low-e glass windows are just a few examples of current market trends.
The two primary methods of production are the float glass process
that produces flat glass, and the glassblowing process, that produces
containers such as bottles and jars. There are significant differences in
the production methods and main ingredients of glass that can lead to
12
Forensic Chemistry 18 (2020) 100223
their classification into end-use categories by measuring their physical
characteristics, optical properties, and major elemental components
[355,363]. Trace elements can be unintentionally incorporated into the
glass during the manufacture, such as zirconium and aluminum that can
leach in the glass melt as a result of the wearing of the refractory bricks
used on the lining of the furnaces. These inadvertent trace additions can
be used to differentiate glass produced in a single plant over short
periods of time [352,355,364].
Glass and its physicochemical properties are complex and diverse
due to its composition, manufacture, distribution, and temporal history.
Therefore, a random match of two broken glasses that originate from
different sources is possible, but unlikely. Indeed, the random match
probability from glass typically encountered in casework has shown to
be relatively small (e.g., less than 0.2% for LA-ICP-MS [69,365,366]
and smaller than 2% for micro-XRF [358].
9.2. Transfer and persistence studies
For glass to become potential questioned evidence, it must transfer
from one object or place to another and then be collected. How easily it
transfers, persists in its transferred state, and remains accessible for
collection is critical information to the glass examiner. Further, in order
for glass to transfer to other objects or individuals, it must be broken. As
a result, the mere presence of glass fragments indicates the person or
object has been in contact with broken glass, either directly or in-
directly [367–370]. Therefore, unbroken glass is not a probable donor
source of broken glass. The information regarding transfer and persis-
tence mechanisms and frequencies of occurrence of background glass is
critical during the investigation of a crime [53,75].
When glass shatters, fragments of all sizes travel in the direction of
the breaking force and in the opposite direction [371]. Therefore, glass
can be deposited on objects located within the vicinity of the incident.
Although transfer and persistence can be variable, there is a degree of
predictability based on information derived from experimental studies
[372–375]. For instance, how much glass is transferred from a broken
window to the vicinity and how much of it remains on the receiver is
influenced by the breaking mechanism and applied force [375–378],
the distance and relative location from the breaking point
[369,370,376], the type and thickness of the glass [53,369,379], the
type of garment worn by an individual [369,372,380–383], the time
elapsed between an incident and the arrest or recovery of glass and
post-breaking activities [53], and the environmental conditions [382].
The number of factors that influence deposition and retention of
glass in or on a questioned item is extensive. Nonetheless, there are
open source statistical packages that are available to examiners to help
estimate transfer probabilities based on information derived from the
case, such as number of fragments recovered, location of the fragments,
and type of window [53,59]. It is also possible to use statistical methods
to predict the number of fragments transferred onto a garment at the
moment of the breaking from the number and distribution of fragments
found on the ground [375]. Glass tends to fall from the receiver over
time, with fragments larger than 5 mm lost more rapidly than smaller
fragments. Fragments smaller than 1 mm could persist up to 8–24 h
depending on the retention properties of the substrate and the post-
breaking activities [369,376].
The prevalence of glass on members of the general population not
involved with glass-breaking activities is also important information.
There are two major types of background survey studies: those con-
ducted on members of the regular population and those conducted on
individuals who have a higher chance of having glass fragments on
their clothing or person, such as suspects who have been implicated in
breaking and entering activities and individuals often exposed to glass.
The recovery of background glass in these population studies
[291,367,384–391] has been focused on shoes, upper and lower
clothing, and headwear. However, recovery methods vary from
shaking, taping, hand-picking, and vacuuming. Some studies focus on
T. Trejos, et al.
surface glass, while others recover glass from pockets and inner areas
[291,384–392]. Differences in experimental designs on regular popu-
lation has contributed to the wide range of results among published
findings [291,367,384–391]. For instance, the reported frequency of
finding glass on the general population ranges from 0 to 44% on the
surface of clothing, from 1.6 to 40% on pockets of clothing, and from 5
to 10% in footwear. The data suggests that geographical and socio-
economic circumstances have an impact on the levels of background
glass; the frequency of occurrence reported in Australia, New Zealand
and Canada are considerably smaller than the frequencies reported in
England and Ireland (<2% to 40%, respectively)
[291,367,386,388,390]. The higher levels reported in the United
Kingdom (UK) are thought to be a result of a particular period of time
when the cities were undergoing high levels of construction. In general,
there is a low probability of finding multiple single-source glass frag-
ments on a person not exposed to broken glass, which provides support
for glass to be examined in forensic casework.
From surveys on suspects who have been arrested by police,
clothing was examined from 689 individuals suspected of criminal ac-
tivity involving broken glass from 405 cases at the UK and Ireland
forensic laboratories [387]. In 68% of the cases, glass was recovered
that was associated to the control source, and in 25% of the cases, glass
that was recovered was different from the control sources. The authors
reported that although it is not unusual to find non-matching glass on
suspects, it is rare to observe more than three non-matching glass
particles from a single source on one individual. On the other hand, a
survey conducted in Australia showed that, from 87 garments from non-
glass casework only 3% bore glass fragments, suggesting that people
involved in an offense not involving broken glass have considerably
lower background glass than offenders suspected of breaking activities
[389]. Similar findings were reported from New Zealand [367].
As expected, individuals exposed to broken glass have higher levels
of glass retention. Low levels of glass were reported on headwear of a
random population in Sydney (2.6%, n = 232) in comparison to gla-
ziers (94.4%, n = 25) [392]. Relatively elevated percentages of glass
were also found on jackets from law enforcement and examiners
dealing with broken glass environments, where 14% of 44 personnel
had 1 to 11 multi-source glass fragments [391].
9.3. Forensic examination protocols
Once glass evidence is received in the laboratory, analysis starts
with a physical examination. When a broken window does not shatter
completely, examiners can assess the fractures to provide information
regarding the direction of the breaking force and sequence of events
[353,355,393]. As one of the first steps in the analysis, the examiner
also evaluates the potential of a physical fit between the known glass
and a questioned fragment. A physical fit of two pieces of glass is
considered the highest level of source attribution in glass examinations
[355]. When a Physical Fit conclusion is made, it is the examiner’s
opinion that the possibility is negligible that the compared glass frag-
ments originated from different broken glass sources.
When a physical fit is not found, the examiner evaluates the physical
features such as thickness and color, optical properties such as RI, and
the elemental composition [302]. Practitioners have developed best
practices and guidelines for forensic examination of glass. The ASTM
E1967 (Automated Determination of Refractive Index), ASTM E2330
(ICP-MS), ASTM E2926 (u-XRF), and E2917 (LA-ICP-MS) are consensus
documents which serve as a model for examination and interpretation
protocols. SWGMAT created a series of guidelines for the collection,
handling, and examination on glass evidence that are available through
the ASTEE website [263]. The OSAC Materials (Trace) Subcommittee is
currently developing an overarching standard guide for forensic glass
analysis and comparison.
During the analysis of glass, both physical and chemical properties
can be used to classify the glass according to their intended use or their
13
Forensic Chemistry 18 (2020) 100223
major composition. Glass is classified by end use as sheet glass (archi-
tectural and vehicle windows), container glass (jars, bottles), decorative
glass (lamps, vases) and specialty glass (fiber optics, optical lens). By
composition, glass is classified as soda-lime (containers, windows),
leaded glass (houseware, decorative glass) and borosilicate glass
(cookware, industrial glass) [353,355,363].
The RI of a glass is a result of its composition and thermal history
and can be measured in many ways, including the Emmons double
variation, dispersion staining, the Becke line method, and automated
immersion methods [355,394]. However, automated oil immersion
refractometry with phase contrast microscopy has been adopted almost
exclusively in forensic laboratories around the world due to the in-
herent advantages of modernization, automated measurements, speed
of analysis, improved precision, and availability of international stan-
dardized methods [355,395–397].
RI can be measured and compared before and after laboratory an-
nealing to enhance source classification, particularly for tempered glass
and non-continuous glass fibers [355,398]. The extent of change in RI
can help distinguish between tempered glass (ΔRI ~ 1 × 10−3) and
non-tempered glass (ΔRI ~ 2 × 10−4 to 6 × 10−4) [53,399–403].
Elemental analysis is one of the most informative steps in glass
comparisons [353,355]. Several studies [404–419] have demonstrated
that the most informative elements in glass comparisons are those
present at minor and trace levels and therefore, an essential attribute of
the analytical method is its sensitivity. The techniques that have found
widespread use in forensic laboratories are SEM-EDS, micro-XRF, ICP-
optical emission spectroscopy (ICP-OES), ICP-MS, LA-ICP-MS, and LIBS.
Three of these methods hold international recognition as standard
methods micro-XRF, ICP-MS and LA-ICP-MS [420–422] and are pre-
ferred for glass comparisons due to their superior analytical perfor-
mance and informing power. A description of the capabilities, ad-
vantages and challenges of these elemental analysis methods can be
found elsewhere [75,352,353,364–366,396,423–430].
As with other trace materials, if any exclusionary differences be-
tween compared samples are found at any point, then examinations can
cease, and it can be concluded that the samples do not share a common
source. By contrast, if the characteristics and properties of the com-
pared items cannot be distinguished at every stage of the glass ex-
amination protocol, then it can be concluded that the glasses could
share a common source. Given that glass is a mass-produced material,
an association implies that the items originated from either the same
source or another source of broken glass with the same properties. The
combined methods used for the examination of glass have demon-
strated to be highly discriminatory, often differentiating glass produced
in different plants or the same manufacturing site over short production
periods [407]. Since there are many plants producing glass over many
years, and glass has to be broken to become a likely source, coincidental
indistinguishable glass items can exist but are infrequent.
9.4. Discrimination and interlaboratory studies
As previously stated, glasses can be discriminated by color, RI,
elemental composition, and other properties. Research has focused on
evaluating the intra- and inter-glass variability of these properties and
the discrimination potential of the techniques, which includes assessing
the analytical performance characteristics, such as accuracy, sensitivity,
selectivity, bias, reproducibility, and repeatability.
9.4.1. Refractive index
The RI of glass is one of the properties carefully monitored by glass
makers as part of their quality control, and as a result, the distribution
of RI across a windowpane or bottle is relatively homogeneous.
Localized homogeneity of the RI of glass is often a reasonable as-
sumption. For instance, the variation in RI across a pane of sheet glass is
reported to be between 2 × 10−5 and 2 × 10−4 for sheet glass
[394,431,432] and 5 × 10−5 to 3 × 10−4 for container glass
T. Trejos, et al.
[433,434]. However, studies have indicated that this is not always the
case, and special sampling procedures should be considered when
dealing with surface versus bulk glass, tempered glass, or glass with
contamination debris [401,435–438].
The RI of glass has been shown to be measured with high precision
and accuracy [397]. The robustness of these measurements was re-
ported, within a one-month period to be 0.24% RSD. Precision (re-
peatability) of the method was in the order of 2 × 10−5. These results
agree with interlaboratory studies that reported repeatability of
1.5 × 10−5 to 2.5 × 10−5 and inter-laboratory reproducibility of
3.8 × 10−5 to 5.0 × 10−5 [439,440].
The measurement and instrumental variations and within-source
variations of RI are often smaller than the variation among different
sources of glass, providing useful discrimination of glass objects.
However, the discrimination of glass by RI is somewhat limited (~3–5%
random match probability), and therefore RI analysis is often combined
with elemental analysis [394,427].
Two of the most recent inter-laboratory studies for RI were co-
ordinated by the ENFSI European Paint and Glass Group and by the
ASTM Interlaboratory Test Group. The RI measurement repeatability
within a lab ranged from 1.5 × 10−5 to 2.5 × 10−5 while inter-lab
reproducibility ranged from 3.8 × 10−5 to 5.0 × 10−5 [439,440].
Although the glass community does not have RI certified references
materials currently available in the market, the reference glasses used
by this collaborative study are accessible to forensic laboratories, and
the data can now serve as an additional quality control assessment.
9.4.2. Micro-XRF
The relative discrimination capabilities of RI, micro-XRF, LIBS and
ICP methods have been evaluated among a set of sheet glass with very
similar refractive indices [358–360,428,441,442]. These studies show
that micro-XRF is suitable for discrimination of glass samples origi-
nating from different plants and from glass manufactured in the same
plant within a short time period (e.g., one month apart, ~2% random
match probability). Although the capabilities of the method to detect
temporal variations in a plant cannot be generalized, they provide va-
luable insight into the method’s potential. Furthermore, micro-XRF has
been adopted by many forensic laboratories and a standard test method
is now available to examiners [358,363,420,428,443,444].
However, unlike RI and ICP methods, the spatial homogeneity of
glass within a single pane has not been directly addressed by micro-XRF
methods. Also, in recent years modern micro-XRF systems are em-
ploying silicon drift detectors instead of the traditional silicon lithium
detectors as they are anticipated to be more sensitive and precise. The
use of synchroton radiation (SR-XRF) has been explored by Japanese
researchers with trace elements detection in the picogram level
[404,434,445].
9.4.3. ICP methods
Numerous publications have described the scientific basis for the
elemental analysis of glass by ICP methods
[405,412–419,434,446–449]. Particular attention has been focused on
the selection of a relevant and informative element list, spatial homo-
geneity and within sample variations of the elemental composition, and
the variation in the elemental signatures among glass from different
sources.
Studies have recommended elements based on their analytical
performance, if they are correlated and how informative or dis-
criminating they are [405,412–416,446–449]. These studies led to the
selection of up to 18 elements included in the ASTM standards E2330
and E2927 that have been demonstrated to provide good discrimination
between different sources of glass and correct associations of glasses
originating from the same source. Some of these elements have been
reported to be highly correlated, but they were still included as their
detection capabilities can vary with ICP configurations.
14
Forensic Chemistry 18 (2020) 100223
The assumptions of spatial homogeneity within a single source is a
fundamental pillar in glass examinations. Several researchers have
reported variations within single panes, architectural and automobile
windows, and containers [365,406,414,417–419,434]. Different
sampling strategies have been applied, including monitoring glass
panes coming out of the production line at different time intervals
[407]. What is important to note, is that over the years, all these
studies have reached consensus that elemental profiles of glass are
homogeneous at a glass-pane level. False exclusion rates have been
reported in the range of 0.3 to 1% for fragments from the same
window and 1.2 to 7.5% for fragments from panes from the same
windshield [69].
Nonetheless, as with any mass-produced materials, examiners have
to be aware of the dynamic nature of manufacturing processes and
market influences. A recent study [408] reported notable heterogeneity
across the thickness of the glass, for some elements, on five float glasses
samples measured by LA-ICP-MS. The authors attributed the anomalous
variation to manufacturing flaws called reams. These heterogeneities
have the potential to cause Type I errors or false exclusions if the known
sample is not adequately characterized. LA-ICP-MS data analysis should
include quality control protocols to identify outliers, and anomalous
high standard deviations for measurements that may be related to
quantitation limits or heterogeneities.
The between-sample variation for glass samples is known
to be larger than the within-sample variation. Variation
between-factories and within-factories in the US, Europe and
Australia at different production intervals have been reported
[359,360,364–366,409–411,414,428,448]. Likewise, within-factory
variability was studied at different time intervals for over 300 glass
fragments collected from single plants over a period of 53 months, in-
cluding approximately 100 samples collected every 15 min for a 24-
hour period [407]. Discrimination capabilities better than 99% have
been consistently reported, from glass originating from different
sources, with experimental random match probabilities in the order of
0.1 to 0.2% [69,359,365,366].
Some of the most significant interlaboratory studies for elemental
analysis of glass using multiple techniques were conducted by the
European Funded NITECRIME group and by the NIJ-funded Elemental
Analysis Working Group. Specifically, the NITECRIME effort was a
valuable step toward the optimization and validation of the LA-ICP-MS
method for glass analysis, with collaboration from users across Europe
and the U.S for a span of 4 years. The final product of this effort con-
stitutes the basis for the current LA-ICP-MS ASTM method
[359,364,413,443]. More recently, interlaboratory studies funded by
NIJ have been focused on the evaluation of interpretation methods for
the forensic analysis of glass and the challenges of sharing and using
glass databases [69,71].
9.5. Collections and databases
To date, the largest glass collections and databases have been built
at and maintained by federal and state forensic laboratories and are
composed primarily of known glass received as control samples in
casework and glass collected from junkyards or manufacturing plants.
In academia, Florida International University hosts one of the largest
collections of glass from different manufacturing plants dating from
1995 to the present. The local laboratory database sizes range from 600
to 2300 sources, including mainly architectural, automotive and con-
tainer glass.
The most extensive current glass databases have been built pri-
marily with RI and ICP data (ICP-MS, ICP-OES, and LA-ICP-MS). It
would be useful for XRF-users to generate more massive collection
datasets that include modern glass compositions to expand the current
knowledge on XRF-elemental profiles of glass and evaluate the poten-
tial of sharing databases among laboratories.
T. Trejos, et al.
The use of relevant databases assists in the validation of the
methods developed for interpretation of glass data. The ASTM methods
include recommendations on comparison criteria when the binary cri-
teria (i.e., whether or not samples can be discriminated) is used, and
these recommendations were formulated based on the performance
evaluation of several comparison methods, error rates estimated
through inter-laboratory testing, and experimental data from databases
[359,365,366,450]. Some of the comparison criteria apply empirically
derived rules that—although not statistically sophisticated—have a
demonstrated fit for purpose and provide low error rates. Nonetheless,
the community continues to evaluate alternative methodologies, in-
cluding the use of LRs and Bayesian networks [53,61,68,69,71,
72,77,451].
There is a vast amount of scientific literature describing the appli-
cation of Bayesian interpretation of forensic glass evidence [50–70]. For
more extensive information regarding Bayesian interpretation of glass
evidence refer to Curran et al. [53,452].
Recent work has been conducted to improve probabilistic evaluations
of glass evidence using LR approaches [68,69,71,72,77,451]. For in-
stance, error rates of the ASTM match criteria and the calibrated LR
continuous approach have been compared using two of the largest ex-
isting glass databases. This study showed that the LRs provided a lower
false exclusion rate than the ASTM criterion, while the ASTM criterion
provided a lower false inclusion rate, although the error rates were still in
the same order of magnitude. This study also demonstrated that using
LRs provides a more objective evaluation of the quality of an association
or exclusion, which is not possible with binary criteria [69,71].
Alternative statistical models based on estimates of means and
covariance matrices have been reported [453]. However, unlike any
other reported algorithms, the discrepancy of error rates between the
modeled data and the ASTM criteria was substantial. A potential ex-
planation of the disagreement is that the author’s modeled elemental
profiles that over-represent false positives. The range of variations se-
lected simulated samples manufactured in the same plant in short time
periods with very similar composition. The inability of the analytical
methods to differentiate this type of samples is well known and docu-
mented and described when the expert explains the “source attribution”
in their conclusions. The range of reported error rates in this study
[453] is also in discrepancy with Bayesian approaches. Moreover, the
authors use an analogy between bullet lead and glass that is entirely
inappropriate given the significant differences between manufacturing
processes, homogeneity, composition, variability studies and most im-
portantly scopes and limitations in the interpretation of glass. As a re-
sult, this study needs further revision and validation before being ap-
plicable in the field.
Another study [454] proposing the use of machine learning algo-
rithms for glass comparisons was recently published. However, the
experimental design also suffers from similar flaws as in [453], se-
lecting samples manufactured in the same plant at short periods to
evaluate error rates. Moreover, this study has severe problems with the
quality of the data used to train the algorithms. In particular, the
measurement of lithium does not meet the ASTM E2927 quality control
criteria, significantly skewing the data and impacting the error rates
reported by the authors. For instance, if the authors would have iden-
tified and removed the outliers in their data, the false-negative rates in
intra-pane comparisons would have dropped from the reported 40.2%
to only 2.4%, which would have been in agreement with the existing
vast literature on glass elemental analysis [455]. Also, the elimination
of poor quality from inter-pane comparisons would have prevented
misleading error rates. A group of scientists submitted a letter to the
editor to raise awareness of the paper's flaws and allow the authors to
amend the errors. Unfortunately, the letter was rejected by the editor
with no reasonable explanation of the basis of the rejection. Therefore,
there is a potential risk of misuse of the paper by stakeholders if they
don't realize the severe deficiencies of the study.
15
Forensic Chemistry 18 (2020) 100223
10. Overall conclusions
This review highlights the scientific foundations for the trace evi-
dence discipline. Current standard methods are based on a broad con-
sensus process that involves the assessment of robustness, accuracy,
repeatability, and reproducibility of the methods within an extensive
community of users. Scientific working groups have contributed sig-
nificantly to the validation of these protocols.
Forensic science can only advance in the right direction with the
combined knowledge and expertise of practitioners, researchers, man-
agers, and statisticians. These strategic partnerships promote the de-
velopment of solutions that are scientifically valid, practical, and effi-
cient. Experimental studies discussed here have been typically designed
with collaborative efforts between forensic experts, academia, and in-
dustry. As a result, most of the existing empirical data have been gen-
erated using casework-like scenarios and samples for a better under-
standing of its significance. We have also provided examples of the risks
of applying correct forensic assumptions but poor statistical inter-
pretation [146,456–458] as well as using valid statistical models but
poor experimental designs based on a wrongful understanding of for-
ensic science [453,454]. Interestingly, these examples raised a flag on
the need for incorporating appropriate levels of editorial or reviewer
forensic expertise in statistical journals.
In the past ten years, some techniques have been investigated for
their application to trace materials including Raman spectroscopy,
LIBS, LA-ICP-MS, liquid chromatography-MS (LC-MS), direct analysis in
real time-MS (DART-MS) and other MS methods. However, the majority
of publications have been focused on further validation of in-
strumentation already available in the forensic laboratories, studies of
transfer and persistence, and application of statistical methods for the
interpretation of evidence, which have provided a valuable foundation
for the discipline of trace evidence.
The trace evidence discipline is diverse and has many facets: a)
many types of materials can be submitted as evidence, each with dis-
criminating characteristics to answer a variety of questions at the
source or activity levels, b) the collection of microscopic trace evidence
requires expertise from crime scene investigators, c) often trace mate-
rials interact with one another and the order/interplay of examinations
has to be determined using case-specific information, d) evidence may
be submitted to different sections within a forensic laboratory, and
when multiple evidence types are examined and reported, the com-
bined value of the evidence is not always clearly articulated, e) the
examination of features and characteristics of the traces is a time-con-
suming process, and f) the composition of trace materials is not static;
therefore, the examiner has to continuously adapt to novel analytical
tools and updated information regarding market changes.
As a result, a sound knowledge base of the different types of trace
evidence is critical to substantiate decisions made during the collection
of evidence, sampling, analysis, and interpretation. Such knowledge
includes an understanding of the material's composition, its relevant
population, and the informative value of the features being evaluated.
Only then are forensic scientists capable of making inferences regarding
where, how, and what to collect at the crime scene, the best strategy to
preserve intersecting forms of evidence, an appropriate sequence of
analytical methodologies, the extrinsic factors that are relevant to as-
sess the weight of the evidence, and the information most significant for
comparison and reconstruction purposes. This paper describes the
foundation of knowledge, training, and resources available for that
decision-making process.
Despite the efforts to date and that are underway, the trace com-
munity seeks continual improvement and will be iteratively working
towards enhancing the perceived value of trace evidence in re-
construction and intelligence investigations; addressing gaps in re-
search; exploring and validating more objective, transparent and
quantitative methods for the interpretation of trace evidence data; and
T. Trejos, et al.
the development and maintenance of larger reference collections and
databases.
This paper provides a review of the current status of trace discipline
in the United States: how the scientific foundations have been estab-
lished and what the current capabilities and limitations are. Particular
emphasis was given to hair, fiber, tape, paint, and glass materials. The
references cited serve as a bibliography supporting the trace evidence
discipline and the interpretation approach currently being developed by
the OSAC Materials (Trace) Subcommittee.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgements
We would like to thank all the outstanding scientists who came
before us. They have provided the scientific foundation for the analyses
and research being conducted in laboratories around the world today.
We would especially like to acknowledge the OSAC Materials (Trace)
Subcommittee for their contributions and to thank Jodi Webb, Candie
Shegogue, and Diana Wright for providing feedback on a draft of this
publication.
This is publication number 19-25 of the FBI Laboratory Division.
Names of commercial manufacturers are provided for identification
purposes only, and inclusion does not imply endorsement of the man-
ufacturer, or its products or services by the FBI. The views expressed are
those of the authors and do not necessarily reflect the official policy or
position of the FBI or the U.S. Government.
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