MODULE FORENSIC CHEMISTRY AND TOXICOLOGY

CHAPTER 10: TRACE EVIDENCE AND DNA ANALYSIS

Objectives:
a) Understand the concept of trace evidence
b) Determine the proper handling of hair sample, fiber evidence
and paint pigment
c) Develop knowledge in Forensic DNA Analysis

❖ TRACE EVIDENCE
o These are any materials left by any suspects or victim which serves as a link to a
crime.
o This includes impressions (tire, footwear, bite, tool mark, etc.), hair, paint, and
fibers, ultraviolet powder, glass and polymers (plastics), pollens, metallic
fragments, and other deposits etc.
o Originally, trace analysis was used by hunters and trackers or sign cutters (people
who read signs and marks of the progress of a journey of people from one place
to another from all kinds of country).

HAIR EVIDENCE

a. HAIR – is an appendage of the skin

that grows out of an organ known as the
hair follicle.

• an individual shed approximately 30

to 100 head hairs each day and grows
1cm per 1 month.

• the transfer of hair during physical

contact, from suspect to victim or crime
scene, can associate the suspect from
the crime that has been committed.

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Types of Transfer
❖ Primary Transfer – hairs transferred directly from the region of the body where they
are growing.
❖ Secondary Transfer – transfer from clothing of the individual.
o The types of hair recovered, and the condition and number of hairs found an impact
on their value as evidence in criminal investigation.
o the length, color, shape, root appearance and internal features are considerable
variable characteristics.
o Three basic parts of the hair: root, shaft and tip.
HAIR ROOT
❖ Root and other surrounding cells contained within the hair follicle is the responsible for
hair production and growth.
❖ Three development stage of hair growth: anagen, catagen, and telogen
o Anagen phase – last up to six years, the root is attached to the follicle for
continued growth, giving the root bulb a flame-shaped. When pulled, anagen hairs
contain follicular tag, a translucent tissue surrounding the hair shaft near the root.
Follicular tag was used for individualization using DNA.
o Catagen phase – typically have an elongated appearance as the root bulb shrinks
in size and being pushed out from the follicle. On this phase, hair continue to grow
at decreasing rate which can last from two to three weeks.
o Telogen phase – begins when hair growth ends, the root takes on a club shaped
appearance. During six to two months period, the hair will be pushed out of the
follicle causing the hair to be naturally shed.

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HAIR SHAFT
➢ Three parts of hair shaft: cuticle, cortex and medulla

❖ Cuticle
o It is a protective coating made of overlapping
scales (appearance of shingles on a roof) that
always point toward the hair tip
o It has a characteristics pattern giving the hair
resistance to chemical decomposition and
retains its structural features over a long time.
o Not useful in individualizing human hair but
can be used for species identification.
o It is composed primarily of protein called
keratin different for every animal. Examined
under compound microscope and stereoscope.

❖ Cortex
o Made of spindled-shaped cells aligned in a
regular array, parallel to the length of the hair.
o Embedded with pigment granules that gives
hair its color.
o The color, shape and distribution of the
granules provide points for forensic comparison.
o Use to categorize human race

❖ Medulla
o Collection of cells having the appearance of
a central canal running through a hair.
o Animal medulla occupy more than half of hair diameter.
o The shape of the medulla can help identify a species. Examples, most animal
humans: cylindrical, Cats: pearl shape, Deer: spherical occupying whole hair shaft
o Four classification include: continuous (mongoloid head hair), interrupted,
fragmented and absent.

• Medullary index – measures diameter of medulla relative to a diameter of the hair shaft
expressed in fraction.
• Human medullary index = less than 1/3
• Animal medullary index = ½ or greater
• Types of animal hairs: guard hairs – coarse outer hair, Fur – finer hair next to the guard
hair, Tactile hairs – whiskers, tail and mane hair.

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o Forensic Identification and Comparison

Standard – considered samples from known source.

Questioned – sample from unknown source.

o Racial origin: negroid hairs is kinky with dense and uneven pigments.
o Body parts: pubic hairs are short and curly with varying shaft diameter and
continuous medulla. Beard are coarse and triangular in cross section with blunt tips
acquired from cutting or shaving.
o Hairs forcibly removed may suggest a violent confrontation. Presence of hair
follicular tissue suggest that hair has been pulled out quickly.
o Age: infant hair is fine, short and fine pigment.
o Sex: presence of dyes and thickness of a hair.
o Environmental alterations can result from exposure to excessive sunlight, wind,
dryness, and other conditions. It is recommended that hair samples be obtained
as soon as possible from suspects and victim.

❖ Sources of Collecting Hair Standards

o Suspect
o Suspect’s clothing
o Suspect’s environment
o Crime scene
o Victim
o Victim’s clothing
o Victim’s environment

PROCESS OF COLLECTING HAIR STANDARDS

Hairs should be pulled one at a time including
the root of the hair using fingers and not with
tweezers or tools to avoid damaging the hair. Place
the hairs in paper envelope and tape all the seams
and label the container with the following:
o Name of the person whose samples
taken
o Date and time of sampling
o Case number
o Name of collector
o Initial and date across the envelope
seal by the collector.

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• Hair standards: 20 to 50 head hairs from various areas of the scalp (crown, sides, front
and back) to ensure all colors and shades of the head hair were collected.
• Pubic hair standards: at least 20-25 pubic hairs by pulling individual hair from various
areas around the pubic region. Pubic hairs are not subject to as much change as head
hairs. A sample taken a year or more after the crime may still be suitable for comparison.
• Facial hair standard: collect at least 20 facial hairs by pulling individual hairs from various
regions of the face to assure a good sampling.

PROCESS OF COLLECTING QUESTIONED HAIR

• Pubic hair combings: usually collected in sexual

assaults cases and from homicide victims. Place a
piece of white paper under the pubic region and
using a new comb, comb through the entire pubic
area to dislodge any foreign hairs or other foreign
materials that may be present.
• Place the used comb onto the paper and fold the
paper around the comb and careful not to lose any
of the dislodge evidence.
• Other question hairs: the hair should be placed
on a piece of white paper and carefully folded
around the hair. The resulted folded paper should
then be placed into a paper envelope and sealed.

FIBER EVIDENCE

Mostly encountered in incidents such as homicide,

assault, or sexual offenses usually found in vehicle in a
hit and run cases, screens or glass broken in the course
of entering and leaving the scene.

• Two General Classifications of Fibers

1. Natural fibers – derived in whole from animal or
plant resources
2. Man-made/ synthetic fiber

• Identification and Comparison

o Cotton has a ribbon-like shape with twists at regular
intervals.

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• Guidelines in collecting fiber standards

o When collecting standards, a sample piece of the cloth or material should be
collected that contains a representative sample of all the colors, textures and
characteristics of the material.
o Each standard should be labeled and placed in a paper envelope, indicating from
where the sample was collected. Label the envelope like in hair evidence.

• Guidelines in collecting questioned fiber

o The use of alternative light source can be very useful tool for locating fiber
evidence. Synthetic fibers especially will fluoresce using proper combination of
light waves and colored goggles.
o Questioned fibers should be collected and handled in a similar manner of methods
describe in collecting standard fiber.

• What is blind search?

o This method is used for searching trace evidence particularly hairs and fibers
which is usually more difficult to find. Blind search is conducted using a clear
packaging tape/ scotch tape 2 inches wide by 1 foot (2” x 1’) long is pressed and
lifted against the back of the drives seat in an overlapping pattern until all residual
hairs and fibers from the entire surface have been lifted onto the tape.
o The tape is then pressed onto a clear sheet of Mylar or a transparency copy sheet
and labeled as to what portion of the seat it was removed from.
o If the tapes become overloaded, use another piece of tape. The transparency
sheet with the tape and hairs can be submitted to the laboratory for analysis.

PAINTS

• A dispersion of pigments in a vehicle and solvents or

in an emulsified vehicle.
• When the paint coats any surface in any manner
such as brushing, the solvents or volatile portion
evaporates. A colored film is developed for the purpose
of decoration and protection.

Types of paint
o Water system paint – water soluble paint, usually diluted with water.
o Emulsion system paint – oil soluble paint, usually diluted with thinner.

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Components of Paints: pigments and additives

suspended in a binder with the aid of a solvent.
• Pigment - color portion
• Binder – support medium, film former which imparts
characteristics of toughness, durability, dry time
• Solvent – evaporates once paint is dry
• Additive – enhance the properties of paint

Common Additives
• Suspending agents – used to prevent settling of pigments. (bentones, aluminum,
stearate).
• Driers – used in alkyds and oils to make them dry faster. (6% Cobalt solution, 24%
Pb solution).
• Anti-skinning agents – used in oils and alkyds to prevent surface skinning in
containers.
• Welling agents – to promote pigment welling and ease of grinding.
• Anti-foaming agent – used in water paint system to minimize foaming and air
entrapment
• Coalescing agent – used in emulsion systems to ensure proper coalescing of latex
particles upon drying
• Thickeners – used in water paints to increase the viscosity of paints
• Viscosity control agents – for solvent paints used to increase viscosity
• Other additives – additives that contribute various properties to paints

Common Paint Evidence

• Automobile, form hit-n-run and collisions,
automobile paint can be distinctive because it
has multiple layer/coats. Modern automotive
finishing consists of at least four coatings:
o Electrocoat primer
o Primer surface
o Basecoat
o Clearcoat

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• Questioned and known specimens are compared side by side under a stereomicroscopic
for color, surface texture, and color layer sequence
Note: layer sequence is very important evidence: forensic scientist will try to match
layers with respect to number and sequence of color. Layer structure alone will not
provide enough information to be individualized to a single source.
• Chemical analysis of the paint’s pigment and binder composition provides further points
of comparison.
• Typically, gas chromatography is used to determine the chemical make-up of the binder
material.
• Infrared spectrophotometry is used to determine the binder composition of paint.
• Elements of the paint pigment can be identified with a number of techniques, including
spectroscopy, neutron activation analysis, and x-ray diffraction.
• Using these techniques, the odds against crime-scene paint originating from another
randomly chosen vehicle is approximately 33,000 to one.

How to examine paint:

a. Microscope most common tool. Look at: color, surface texture,
and layer sequence when identifying relatedness/ origin.
Problem: If layering is not complex, cannot
identify single source or common
origin.
b. Chemical properties – look at binder chemically using
gas chromatography or infrared radiation spectrum.

Collection/ Analysis of Paint Evidence:

• Use tweezers, scalpel or paper to obtain paint evidence. Store it in paper, glass, or plastic
vials. If in/on a garment, submit whole garment – DO NOT TRY TO REMOVE.

Comparison – need control sample for area near damaged area. Approximately ½ square area.
If on a tool, package whole tool with control samples. Do not collect paint from an impression,
only adjacent areas.

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TOOL MARKS AND SERIAL NUMBER RESTORATION

TOOL MARK
• A tool is an instrument or object capable of making
a mark on another object. While a tool mark is defined as
any impression, compression, cut, gouge, scratch,
indentation, or any other making left in an object by another
harder object or instrument.
• Application of tool mark: knife marks on bone,
Fractured knife blades, Homemade explosive devises,
Tool mark evidence crimp marks on detonators, cut marks on wire and pry
of lockpicking marks on a window or door.
• What information that tool mark can provide?
The type of tool, shape of the cutting edge of a blade, blade width, color of the tool
(based on the paint transfer from the tool). Class characteristics of the tool., unique
identifying marks.

Types of Tool Marks

• Embossed/ impression Marks – produced by perpendicular force acting against
an object. (e.g. punches, hammers, and some gripping tools)
• Striated scrape marks – made by moving the tool laterally across the object.
(e.g. flat-bladed tools such as crowbars, pry bars and screwdrivers.)
• Shearing or pinching marks – occur when the object is caught between
opposing forces of two cutting actions. (e.g. scissors cause shearing, pliers and
bolt cutters cause pinching).

Class and Individual Characteristics of Tool Mark Evidence

Class characteristics are common to a group of similar objects or by manufacturer.

Screwdrivers have many different heads (flat head and Philips head) so have different class
characteristic.
Between flat head screwdriver, they vary with blade thickness and wide or narrow heads.
Thus, every manufactured design constitutes a different class characteristic.
Individual characteristics are accidental or unintentional marks or defects left on the tool
through the use of the tool (worn out). Same brand of screwdriver but the other one is worn out
produced different marks thus making an individual characteristic.

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General Guidelines in Collection and Packaging Tool Mark Evidence

• Protect the surface of the tool mark

• The best method is to collect the whole object
bearing the mark
• Separate the package of tool and tool mark to
protect the tool, possible fingerprints and trace
material.
• If the object with tool mark is too big (door or
window), cut around the tool mark or cast the tool
mark with photographing. Use casting materials
that do not shrink like silicone.
• Collect the suspected tool only.
• Protect the part of the tool that produced tool mark.

Laboratory Examination of Tool Mark

• Tool mark comparison begins at the crime scene.
Object with tool mark and suspected tool are collected
by the crime scene investigator.
• Tool mark examiner conduct physical view of the
specimen by nothing manufacturer, model,
dimensions, any defect or damage visible on the tool
and make visual examination of the tool mark.
• Evaluate the type of tool marks as embossed, striated
or shearing.
• Measure the lengths and widths
• Create test marks with the tool. Should be reproduced
similar with the evidence material and created in a manner similar to the way the
questioned tool mark was created. A softer material can be used if the evidence material
is hard damaging the tool changing individual characteristics of the tool.
• The marks are microscopically compared for unique and repetitive mark on both standard
and questioned specimen.

Typically Results of Tool Mark Comparison: positive identification, an elimination, inconclusive

results.

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• SERIAL NUMBER RESTORATION

Serial number – a number of related

or similar objects arranged in order or
success

ULTRAVIOLET EXAMINATION

Ultraviolet radiation
• Also known as black light which is the region of
electromagnetic spectrum between 156 and 4000 A.

I. PROPERTIES OF LIGHT

Light is described as the combination of particles and

waves. Light waves consist of perpendicular, oscillating
electric and magnetic field.

Absorption of light

❖ When a molecule absorbs a photon from light, the energy of the molecule increases. When
a photon is emitted, the energy decreases.
▪ Ground state – the lowest energy state of molecule
▪ Excited state – the highest energy state of molecule

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Electromagnetic Spectrum

Table 1: Electromagnetic spectrum and its corresponding type of absorption.

EMR Types of Absorption

Radio wave none

Microwave Rotational motion

Infrared Vibrations of molecules

Visible Electron excitation

Ultraviolet Electron

X-Ray Break bonds and ionize molecules

Gamma ray Break bonds and ionize molecules

FORENSIC DNA ANALYSIS/

DNA FINGERPRINTING

• What is DNA?
o Inside the trillion cells of the human body are strands of genetic material called
chromosomes. Arrange along the chromosomes are beads on a thread of 100,000
genes.
o DNA is a type of substance called polymer, are very large molecule composed of
series of repeating units called nucleotides.
o Nucleotide is composed of a sugar molecule, a phosphorus containing group and
a nitrogen containing molecule called base.
o S designates the sugar component and B is the phosphate group to form a
backbone of DNA. Watson and Crick discovered that the DNA is actually two
strands and coiled called double Helix.
o Blood contains DNA comma and depending on the size of the strain and its
condition a forensic scientist may be able to get enough information to obtain a
highly probable much of a suspect with the evidence. The two techniques are
heavily used by forensic scientists in evaluating DNA evidence from blood or other
body tissues - Polymerase Chain Reaction or PCR in Variable Number Tandem
Repeats or VNTR's

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References:
Daniel C Harris, Quantitative Chemical Analysis, 1999
Terrorism Investigation Course. Lesson 18: Evidence Collection and Preservation of tool marks &
Lesson 20: Evidence Collection and Preservation of hairs and fibers, December 2002.
Drum, James A, The Fundamentals of Paint Technology, Union Oil Company, California.

LINKS

TOPIC LINK FOR VIDEO

Inside the Crime Lab: Trace
https://www.youtube.com/watch?v=bqGrlcDKWKA
Evidence Unit
Microscopic Comparison of Hair
https://www.youtube.com/watch?v=e3ZlA5GoFVw
and Fiber Evidence
Microscopic Comparison of
https://www.youtube.com/watch?v=ArMOsjqCsJw
Toolmark Evidence

https://www.youtube.com/watch?v=rPYTQlFNk5Q&list
Firearms Evidence Guidelines
=TLPQMjYwNDIwMjDXGHcV0T4owA&index=2

Restoring Obliterated Serial

https://www.youtube.com/watch?v=ld4hJmxP9Zk
Numbers
DNA Fingerprinting | https://www.youtube.com/watch?v=7onjVBsQwQ8

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