FORENSIC

CHEMISTRY
LAB
TOPICS

Test Methods

Experiments

Practice Test
 TEST METHODS IN
FORENSIC CHEMISTRY
PARAFFIN TEST
•

•
• DIFFERENT NAMES FOR THIS TEST : LUNGE'S TEST; DIPHENYLAMINE TEST; DERMAL
NITRATE TEST.

• THE BASIS OF THIS TEST IS THE PRESENCE OF NITRATES IN THE GUNPOWDER RESIDUE.

• THE TEST IS DONE BY APPLYING WARM MELTED PARAFFIN WAX ON THE HANDS OF
THE ALLEGED SHOOTER AND CAUSES THE PORES OF THE SKIN TO OPEN AND EXUDE
THE PARTICLES OF GUNPOWDER RESIDUE.

• THESE PARTICLES ARE EXTRACTED BY THE PARAFFIN CAST (MADE OF PARAFFIN

WAX AND ABSORBENT COTTON) AND WILL APPEAR AS BLUE SPECKS,
WHEN DIPHENYLAMINE (DPA) REAGENT IS ADDED ON THE CAST.

• THE SPECKS ARE GENERALLY LOCATED ON THE AREA OF THE THUMB AND
FOREFINGERS.
• TIME ALLOTMENT FOR PERFORMING A PARAFFIN CAST : 72 HOURS (3 DAYS)
FROM THE TIME OF THE ALLEGED SHOOTING INCIDENT.

• NITRATES FROM THE PORES CANNOT BE REMOVED BY ORDINARY WASHING

OF THE SKIN AND STAY FOR 72 HOURS.
 • FERTILIZER
• EXPLOSIVES
SUBSTANCES
• TOBACCO
GIVING FALSE
POSITIVE RESULTS • URINE
• FOOD SAMPLES
• CIGARETTES
 • TYPES OF CALIBER OF AMMUNITION
• USE OF GLOVES
• AGE OF THE GUN
FACTORS FOR
• DIRECTION OF FIRING
FALSE NEGATIVE
RESULTS • WIND DIRECTION AND VELOCITY
• HUMIDITY
• TEST TAKEN WHEN 72 HOURS HAS
PASSED
• PARAFFIN TEST IS NOT CONCLUSIVE EVIDENCE. I T IS JUST AN EVIDENCE
THAT INTENDS TO SUPPORT THE TESTIMONY OF A WITNESS.
 • I T CAN ASSOCIATE AN INDIVIDUAL
WITH A FIREARM BUT NOT SUCCESSFUL
IN IDENTIFYING THAT PERSON AS THE
SHOOTER.
LIMITATION OF THE
PARAFFIN TEST • I T IS ALSO POSSIBLE, BUT VERY
UNLIKELY, THAT RESIDUE WOULD BE
DEPOSITED ON HANDS BY OTHER
MEANS.

• NON SPECIFICITY
 • HAIR IS PRESENT ON MANY DIFFERENT
REGIONS OF THE BODY . EACH REGION, SUCH
AS THE HEAD, PUBIC AREA, CHEST, AXILLAE,
AND LIMBS, HAS HAIRS WITH MICROSCOPICAL
CHARACTERISTICS ATTRIBUTABLE TO THAT
REGION.
• DURING THE ANAGEN PHASE, THE HAIR IS
HAIR ANATOMY & ACTIVELY GROWING, AND MATERIALS ARE
GROWTH DEPOSITED IN THE HAIR SHAFT BY CELLS FOUND
IN THE FOLLICLE
• IN THE TELOGEN PHASE, THE FOLLICLE IS
DORMANT OR RESTING.
• THE TRANSITION PERIOD BETWEEN THE
ANAGEN AND TELOGEN PHASES IS REFERRED
TO AS THE CATAGEN PHASE.
HAIR ANATOMY &
GROWTH • THE MICROSCOPICAL APPEARANCE OF
THE ROOT AREA WILL ALLOW FOR THE
DETERMINATION OF THE GROWTH PHASE.

• NATURALLY SHED HAIRS, SUCH AS A HEAD

HAIR DISLODGED THROUGH COMBING,
DISPLAY UNDAMAGED, CLUB-SHAPED
ROOTS
 HAIR FORCIBLY
REMOVED

• A HAIR FORCIBLY REMOVED

FROM THE SCALP WILL
EXHIBIT STRETCHING AND
DAMAGE TO THE ROOT
AREA.

• FORCIBLY REMOVED HAIRS

MAY HAVE TISSUE ATTACHED
 • HEAD HAIRS ARE USUALLY THE LONGEST HAIRS
ON THE HUMAN BODY .
• THEY ARE CHARACTERIZED AS HAVING A
HEAD HAIRS UNIFORM DIAMETER AND, OFTEN, A CUT TIP.
• HEAD HAIRS CAN APPEAR UNCUT, WITH
TAPERED TIPS BUT ARE MORE OFTEN CUT WITH
SCISSORS, RAZORS, OR CLIPPERS.
• IN GENERAL THESE HAIRS ARE SUBJECT TO
MORE ALTERATION THAN HAIRS FROM OTHER
BODY AREAS.
• ALTERATIONS TO THE NATURAL APPEARANCE
OF HAIR INCLUDE USE OF HAIR DYES, RINSES,
PERMANENTS, FROSTS, AND OTHER CHEMICAL
APPLICATIONS. ENVIRONMENTAL ALTERATIONS
CA
A RAZOR CUT HAIR A SPLIT HAIR
PUBIC HAIRS
• PUBIC HAIRS ARE GENERALLY COARSE
AND WIRY IN APPEARANCE.

• THEY EXHIBIT CONSIDERABLE DIAMETER

VARIATION OR BUCKLING AND OFTEN
HAVE A CONTINUOUS TO DISCONTINUOUS
MEDULLA. WHILE TAPERED TIPS ARE
COMMON, THESE HAIRS MAY ALSO BE
ABRADED OR CUT.
A HAIR SHOWING
BUCKLING
 • PUBIC HAIRS ARE NOT SUBJECT TO AS
MUCH CHANGE AS HEAD HAIRS OVER
PUBIC HAIRS TIME, AND BECAUSE OF THIS, A SAMPLE
TAKEN A YEAR OR MORE AFTER A CRIME
MAY STILL BE SUITABLE FOR MEANINGFUL
COMPARISON PURPOSES.

• I T IS RECOMMENDED THAT A KNOWN

PUBIC HAIR SAMPLE BE OBTAINED AS SOON
AS POSSIBLE AFTER A CRIME AND SHOULD
CONTAIN AT LEAST 25 FULL-LENGTH HAIRS
TAKEN FROM DIFFERENT AREAS OF THE
PUBIC REGION.
FACIAL HAIR • FACIAL HAIRS ARE MORE COMMONLY
CALLED BEARD HAIRS OR MUSTACHE
HAIRS.

• THESE HAIRS ARE COARSE IN APPEARANCE

AND CAN HAVE A TRIANGULAR CROSS
SECTION. HEAVY SHOULDERING OR
TROUGHS IN THE HAIR ARE OBSERVED
UNDER MAGNIFICATION.

• OTHER CHARACTERISTICS INCLUDE A WIDE

MEDULLA AND A RAZOR-CUT TIP.
LIMB HAIR
• HAIRS FROM THE LEGS AND ARMS
CONSTITUTE LIMB HAIRS.

• THESE HAIRS ARE SHORTER IN LENGTH,

ARC-LIKE IN SHAPE, AND OFTEN ABRADED
OR TAPERED AT THE TIPS.

• THE PIGMENT IN LIMB HAIR IS GENERALLY

GRANULAR IN APPEARANCE, AND THE
MEDULLA IS TRACE TO DISCONTINUOUS
 • A HUMAN HAIR CAN BE ASSOCIATED WITH
A PARTICULAR RACIAL GROUP BASED ON
ESTABLISHED MODELS FOR EACH GROUP.

• FORENSIC EXAMINERS DIFFERENTIATE

BETWEEN HAIRS OF CAUCASOID
RACIAL (EUROPEAN ANCESTRY), MONGOLOID
DETERMINATION (ASIAN ANCESTRY), AND NEGROID
(AFRICAN ANCESTRY) ORIGIN, ALL OF
WHICH EXHIBIT MICROSCOPIC
CHARACTERISTICS THAT DISTINGUISH ONE
RACIAL GROUP FROM ANOTHER.

• HEAD HAIRS ARE GENERALLY CONSIDERED

BEST FOR DETERMINING RACE
CAUCASOIDS • HAIRS OF CAUCASOID OR CAUCASIAN
ORIGIN CAN BE OF FINE TO MEDIUM
COARSENESS, ARE GENERALLY STRAIGHT
OR WAVY IN APPEARANCE, AND EXHIBIT
COLORS RANGING FROM BLONDE TO
BROWN TO BLACK.

• THE HAIR SHAFTS OF CAUCASIAN HAIRS

VARY FROM ROUND TO OVAL IN CROSS
SECTION AND HAVE FINE TO MEDIUM-
SIZED, EVENLY DISTRIBUTED PIGMENT
GRANULES.
 • HAIRS OF MONGOLOID OR ASIAN ORIGIN ARE
REGULARLY COARSE, STRAIGHT, AND CIRCULAR IN
MONGOLOIDS CROSS SECTION, WITH A WIDER DIAMETER THAN
THE HAIRS OF THE OTHER RACIAL GROUPS.
• THE OUTER LAYER OF THE HAIR, THE CUTICLE, IS
USUALLY SIGNIFICANTLY THICKER THAN THE
CUTICLE OF NEGROID AND CAUCASIAN HAIRS,
AND THE MEDULLA, OR CENTRAL CANAL, IS
CONTINUOUS AND WIDE.
• THE HAIR SHAFT, OR CORTEX, OF MONGOLOID
HAIR CONTAINS PIGMENT GRANULES THAT ARE
GENERALLY LARGER IN SIZE THAN THE PIGMENT
GRANULES OF CAUCASIAN HAIRS AND WHICH
OFTEN APPEAR TO BE GROUPED IN PATCHY AREAS
WITHIN THE SHAFT.
• MONGOLOID HAIR CAN HAVE A CHARACTERISTIC
REDDISH APPEARANCE AS A PRODUCT OF ITS
PIGMENT.
 • HAIRS OF NEGROID OR AFRICAN ORIGIN
NEGROIDS
ARE REGULARLY CURLY OR KINKY, HAVE A
FLATTENED CROSS SECTION, AND CAN
APPEAR CURLY, WAVY, OR COILED.

• NEGROID PIGMENT GRANULES ARE

LARGER THAN THOSE FOUND IN
MONGOLOID AND CAUCASIAN HAIR AND
ARE GROUPED IN CLUMPS OF DIFFERENT
SIZES AND SHAPES.
• TWISTING OF THE HAIR SHAFT, KNOWN AS
BUCKLING, CAN BE PRESENT, AND THE HAIR
SHAFT FREQUENTLY SPLITS ALONG THE LENGTH.
BIOLOGICAL OR
ENVIRONMENTAL
ALTERATIONS

• WHEN HAIRS ORIGINATE FROM A BODY IN

A STATE OF DECOMPOSITION, A DARK
BAND MAY APPEAR NEAR THE ROOT OF
THE HAIR. THIS CHARACTERISTIC HAS BEEN
LABELED A POSTMORTEM ROOT BAND

POST MORTEM ROOT BANDS

DYED, BLEACHED & NATURAL HAIR
 • COLOR: HUMAN HAIR IS MORE
ANIMAL HAIR VS. CONTINUOUS IN COLORING VERSUS
HUMAN HAIR PET HAIR IS OFTEN BANDED.

• PIGMENT CONCENTRATION: HUMAN

HAIR HAS EVEN DISTRIBUTION OF
PIGMENT THROUGHOUT CORTEX, WHILE
ANIMAL HAIR IS MORE CONCENTRATED
TOWARD THE CENTER/MEDULLA AREA.

• MEDULLARY INDEX: ANIMAL HAIR

RATIO OF MEDULLA TO SHAFT
DIAMETER IS MUCH GREATER THAN
HUMANS—1/2 OR MORE
 • A FIBER IS THE SMALLEST UNIT OF A TEXTILE
MATERIAL THAT HAS A LENGTH MANY TIMES
GREATER THAN ITS DIAMETER.
• FIBERS CAN OCCUR NATURALLY AS PLANT
AND ANIMAL FIBERS, BUT THEY CAN ALSO
BE MAN-MADE.
• A FIBER CAN BE SPUN WITH OTHER FIBERS
FIBER TO FORM A YARN THAT CAN BE WOVEN
OR KNITTED TO FORM A FABRIC.
• THE TYPE AND LENGTH OF FIBER USED, THE
TYPE OF SPINNING METHOD, AND THE TYPE
OF FABRIC CONSTRUCTION ALL AFFECT THE
TRANSFER OF FIBERS AND THE
SIGNIFICANCE OF FIBER ASSOCIATIONS.
 • CONSIDERING THE VOLUME OF TEXTILES
PRODUCED WORLDWIDE EACH YEAR, THE
NUMBER OF TEXTILES PRODUCED WITH ANY ONE
FIBER TYPE AND COLOR IS EXTREMELY SMALL.
• THE LIKELIHOOD OF TWO OR MORE
MANUFACTURERS EXACTLY DUPLICATING ALL OF
THE ASPECTS OF THE TEXTILE IS EXTREMELY
REMOTE.
FIBER
• UNLIKE FINGERPRINTS OR DNA, THEY CANNOT
PINPOINT AN OFFENDER IN ANY DEFINITIVE
MANNER.
• THERE MUST BE OTHER FACTORS INVOLVED, SUCH
AS EVIDENCE THAT THE FIBERS CAN
CORROBORATE OR SOMETHING UNIQUE TO THE
FIBERS THAT SET THEM APART
NATURAL VS.
MANUFACTURED
• NATURAL FIBERS COME FROM
PLANTS (COTTON) OR ANIMALS
(WOOL).
• M ANUFACTURED FIBERS ARE
SYNTHETICS LIKE RAYON, ACETATE,
AND POLYESTER, WHICH ARE
MADE FROM LONG CHAINS OF
MOLECULES CALLED POLYMERS.
NATURAL FIBER
• M ANY DIFFERENT NATURAL FIBERS
ORIGINATING FROM PLANTS AND
ANIMALS ARE USED IN THE
PRODUCTION OF FABRIC.

• COTTON FIBERS ARE THE PLANT

FIBERS MOST COMMONLY USED IN
TEXTILE MATERIALS
OTHER PLANT FIBER

• NATURAL FIBERS COME FROM

PLANTS (COTTON) OR ANIMALS
(WOOL).
• M ANUFACTURED FIBERS ARE
SYNTHETICS LIKE RAYON, ACETATE,
AND POLYESTER, WHICH ARE
MADE FROM LONG CHAINS OF
MOLECULES CALLED POLYMERS.
ANIMAL FIBER
• WOOL IS THE MOST FREQUENTLY USED IN
THE PRODUCTION OF TEXTILE MATERIALS
• AND THE MOST COMMON WOOL FIBERS
ORIGINATE FROM SHEEP.
• FINER WOOLEN FIBERS ARE USED IN THE
PRODUCTION OF CLOTHING
• COARSER FIBERS ARE FOUND IN
CARPET.
• FIBER DIAMETER AND DEGREE OF SCALE
PROTRUSION OF THE FIBERS ARE OTHER
IMPORTANT CHARACTERISTICS.
MANMADE FIBER
• POLYESTER AND NYLON FIBERS
ARE THE MOST COMMONLY
ENCOUNTERED MAN-MADE
FIBERS, FOLLOWED BY ACRYLICS,
RAYONS, AND ACETATES .

• THERE ARE ALSO MANY OTHER

LESS COMMON MAN-MADE
FIBERS.
 • FIBERS SHOULD BE FIRST EXAMINED
WITH A STEREOMICROSCOPE.
PHYSICAL FEATURES SUCH AS CRIMP,
LENGTH, COLOR, RELATIVE DIAMETER,
LUSTER, APPARENT CROSS SECTION,
DAMAGE, AND ADHERING DEBRIS
MICROSCOPY SHOULD BE NOTED.

• FIBERS ARE THEN TENTATIVELY

CLASSIFIED INTO BROAD GROUPS SUCH
AS SYNTHETIC, NATURAL, OR
INORGANIC. I F THE SAMPLE CONTAINS
YARNS, THREADS, OR SECTIONS OF
FABRIC, CONSTRUCTION SHOULD BE
RECORDED
 • TEST TO IDENTIFY FABRIC, A SIMPLE
BURN TEST CAN BE DONE TO
BURN TEST DETERMINE IF THE FABRIC IS A NATURAL
FIBER, MAN MADE FIBER, OR A BLEND
OF NATURAL AND MAN MADE FIBERS.
BURN TEST
FABRIC FLAME ODOR ASH COMMENTS
QUALITY QUALITY
flame w ill self extinguish if
blackish; flame source is remov ed;
WOOL orange color;
sputtery
burning hair or feathers turns to pow der when crushed no smoke

burns more easily than w ool

burns slowly burning hair or feathers but will self extinguish if flame
SILK grayish; turns to pow der when
crushed
source is remov ed

burning paper or leav es

COTTON yellow to orange color; steady
flame
grayish, fluffy slow burning ember

takes longer to ignite than

yellow to orange color; steady burning paper or leav es similar to cotton cotton but otherwise v ery
LINEN flame similar
BURN TEST
FABRIC FLAME ODOR ASH COMMENTS
QUALITY QUALITY
ember will continue to glow
fast orange flame burning paper or leav es after flame source is
RAYON almost no ash remov ed

orange flame; sw eet or fruity smell hard shiny black bead black smoke
sputtery
POLYESTER
will continue to burn after
burns and melts; sizzly acidic or v inegary hard black bead flame source is remov ed
ACETATE

burns slow ly and melts; blue

base and range tip; no burning celery hard grayish or brow nish self extinguish if flame source
NYLON smoke bead is remov ed

w ill continue to burn after

burns and melts; flame source is remov ed
ACRYLIC white orange tip; acrid black hard crust
no smoke
PRINCIPLE OF
KASTLE – MEYER TEST
• Blood can usually be located by the visual appearance of the stain (red-brown color).
• This is augmented by testing with presumptive tests for blood, such as the
phenolphthalein (Kastle-Meyer) test.
• This test relies on the peroxidase-like activity of the heme groups associated with the
hemoglobin contained in red blood cells.

• In the presence of hydrogen peroxide, this peroxidase-like activity will catalyze the
oxidation of phenolphthalin, which is colorless in solution, into phenolphthalein resulting in
a pink colored solution. The presence of a pink color is a positive test result indicating the
presumptive presence of blood.

AH2 + H2O2 –[heme] A + 2H2O phenolphthalin phenolphthalein (colorless) (pink)

TEST RESULTS
LIMITATIONS OF
KASTLE – MEYER TEST
• The phenolphthalein test is used for the presumptive identification of blood. A
confirmatory test may be required to positively identify a stain as containing
blood.

• The phenolphthalein test depends upon the catalytic peroxidase-like activity of

the heme group in hemoglobin. Because hemoglobin is used for oxygen and
carbon dioxide transport in other organisms, this test will react with blood from
animals as well as humans.

• Insufficient sample quality and/or quantity could limit the development of a

positive reaction.

• A color change must be observed within 15 seconds due to the oxidative nature
of the reaction. An unlimited detection time could lead to a false positive
reaction.