Forensic Chemistry and Toxicology

Part 1 – Forensic Chemistry

I. INTRODUCTION TO FORENSIC CHEMISTRY AND TOXICOLOGY

Forensic Science

 Application of scientific techniques and principles that provides evidence for administration of
justice

CHEMISTRY

 The branch of chemistry which deals with the identification of the substances of which matter is
composed.

Forensic Chemistry

 The branch of chemistry which deals with the application of chemical principles in the solution of
problems that arise in connection with the administration of justice
 The use of chemistry to support civil and criminal litigation

Scopes of Forensic Chemistry

 It includes chemical side of criminal investigation

 It includes the analysis of any material the quality of which may give rise to legal proceeding
 It has invaded other branches, questioned documents, dactyloscopy, and photography

Branches of Forensic Chemistry

 Dangerous drugs
 Explosive examination and principles
 Gunshot residue
 Forensic toxicology
 Blood alcohol and drug test
 Examination of counterfeit products
 Arson investigation
 Macro etching
 Bullet trajectory
 Tools and other marks

Physical Evidence

Are articles and materials which are found in connection with an investigation and which aid in
establishing the identity of perpetrator or the circumstances under which the crime was committed or which in
general assist in the prosecution of criminal.

History of Forensic Chemistry and Toxicology

 Greeks, Romans and Egyptians – known to be the early civilizations who used poisons for
murder and execution
 Democritus – was probably the first chemist to study poisons, and he communicated some of his
findings to Hippocrates
 James marsh (1836) – a Scottish chemist, who first use toxicology (arsenic detection) in a jury
trial. Developed the Marsh Test
 Jean Sevais Stas (1851) – a chemistry professor from Brussels, Belgium who first to successfully
identify vegetables poisons in body tissue. Developed the Stas Method (primary isolation technique
for analysis of non-volatile organic substance)
 Douglas M. Lucas – in Canada, he described the application of gas chromatography to the
identification of petroleum products
 Socrates – was forced to drink hemlock (conium genus one of the highly poisonous plant) for
corrupting the youth of Athens.
 Cleopatra commited suicide through the bite of an asp snake
Mathieu Joseph Bonaventura Orfila

 a. Father of forensic chemistry and toxicology

Spanish toxicologist, chemist medical professor in France

b. Published a ground breaking text on toxicology in 1814. His book, a “Treatise of General Toxicology”
(loosely translated from the longer French title) was the first successful attempt to classify poisons as corrosive,
astrigents, acrids, stupefying or narcotics, narcotic acrids or putrefacients

c. He also testified in court as expert witness against madam marie Lafarge, after poisoning her husband,
Charles Lafarge with arsenic using marsh test.
-
 Blandy Murder Case- “First Murder Trial” to show toxicological testimony in 1752 England.
 Mustard Gas- a highly poisonous gas that was used during WW I in 1914 by Germans.

PHILIPPINES
 In the Philippines, the first public recognition of the value of science was made when the position of
“Medicos Titulares” was created in the Philippines by virtue of the Royal Decree No. 188 of Spain dated
March 31, 1876.
 For every province, a Forensic Physician was assigned to perform public sanitary duties and at the same
time medico-legal aids to the administration of justice.
 December 15, 1884, Gov. General Joaquin Javellar created committee to study the mineral waters of
Luzon and appointed Anacleto del Rosario as chemist.
 At present, there are five distinct laboratories in the Philippines,
o Technical Services of the National Bureau of Investigation
o PDEA
o PNP Crime Laboratory
o Public Attorney’s Office
o UP- College of Biology

Forensic Chemist
o The one who analyze evidence from the crime scene and derives a conclusion based on the test
undertaken.

ROLE OF FORENSIC CHEMIST IN SCIENTIFIC CRIMINAL INVESTIGATION

A. Performs analytical examination of different controlled substances such as dangerous drugs and
explosive ingredients.
B. Conduct gunshot residue examinations.
C. Identifies different peculiarities from trace evidence.
D. Examiners body fluids for any degree of intoxication of alcohol, drugs and poisons.
E. Analyzes fake products for unfair competition.
F. Prepares technical reports, prepares findings for court presentation and testifies concerning
scientific facts.

STAGES IN THE PRACTICE OF FORENSIC CHEMISTRY

1. Collection or Reception of the specimen to be examined
o The chemist should personally collect all the specimens necessary for the examination.

Principles in the Collection and Transportation of Physical Evidence

a. Sufficiency of the specimen
b. Standard for comparison
c. Maintenance for individuality
d. Labeling and Sealing
2. Actual Examination of Specimen
3. Communication of Results
4. Court Appearance

GOLDEN RULES IN THE PRACTICE OF CHEMISTRY

a. Go Slowly
o Take all the time to make the test complete
b. Be thorough
o Make careful observation and conduct all sufficient analysis before releasing a result to prevent
mistakes.
c. Take Notes
 o Used laboratory notebook, photograph, voice recorder or other means to record all your
observations.
d. Consult Others
o Consulting others who already handles similar case who could help speed up the investigation.
e. Use imagination
o Be intuitive on the possible outcome of the analysis being conducted.
f. Avoid Complicated Theories
o Explain the assess and interpret the result of the analysis in a simple manner.

FACTORS CONTRIBUTING TO THE LOSS OF PHYSICAL EVIDENCE

1. Lack of Precautions Preventing Tampering of Specimen
2. Failure in Preservation
3. Failure in Transport of Specimen
4. Failure in Identifying specimen
5. Improper packing of Specimen

TYPES OF EXAMINATION USED IN FORENSIC CHEMISTRY (ANALYTICAL CHEMISTRY)

1. Qualitative Examination- process used to determine and identify the chemical properties of an unknown
substance.
2. Quantitative Examination- process used to determine the amount of a specific substance such as
measuring the size, weight, mass and length of a specimen.

METHODS OF ANALYSIS IN FORENSIC SCIENCE

1. Wet Method- requires much time and effort
2. High-Precision Method- this refers to the utilization of UV and IR Spectrometry.

TECHNIQUES USED IN FORENSIC CHEMISTRY

A. MICROSCOPY- this refers to the technical field of using microscopes to view samples and objects that
is not visible to the naked eye.
B. PHOTOGRAPHY- refers to the process of duplicating the image of an evidence for its preservation.
C. INVISIBLE RAYS
D. CHROMATOGRAPHY- biophysical technique that enables the separation, identification, and
purification of the components of a mixture for qualitative and quantitative analysis.
E. ELECTROPHORISIS- refers to laboratory technique used to separate DNA, RNA, or protein molecules
based on their size and electrical charge.
F. SPECTROGRAPHY- this refers to the technique of using spectrograph, an optical device for breaking
light down into the spectrum.
G. NEUTRON ACTIVATION ANALYSIS- refers to a nuclear process used for determining the
concentrations of elements in a vast amount of materials.
H. X-RAY DIFFRACTION (XRD)- analytical technique primarily used for phase identification of a
crystalline material and can provide information on unit cell dimension. The material is finely ground
and homogenized.
I. DNA TYPING or DNA PROFILING- used to establish identity, parentage, family relationships and
appropriate matches.
J. ATOMIC ABSORPTION SPECTROMETRY (AAS)- refers to an analytical technique that measures
the concentrations of elements. Makes use of the wavelengths of light specifically absorbed by an
element.

CHARACTERISTICS OF TOOLS AND TECHNIQUES USED IN FORENSIC SCIENCE

1. SENSITIVITY
2. SPECIFICITY
3. RAPIDITY

PRINCIPLE USED IN FORENSIC CHEMISTRY

a. Law of Individuality- every object, natural or man-made has unique characteristics and not duplicated
in any other object.
b. Law of Progressive Change- everything changes with the passage of time.
c. Principle of Comparison- only “likes” can be compared.
d. Principle of Analysis- Analysis of two or more sets to understand any difference.
e. Law of Probability- All identifications, definite or indefinite, are made consciously or unconsciously
based on probability.
I. BLOOD AND BLOOD ANALYSIS

o About 6 quarts of blood in a man of average sized man.

o Made up of formed elements called plasma. Which red cells or erythrocytes, white blood cells or
leucocytes and platelets.
o Added with oxalate settles down the formed elements leaving the plasma.
o About 55% of the blood is plasma and about 92% is water.
o 10% of plasma are proteins that consist of albumin, globalines, and fibrogen.
o If the blood clots, a strawyellow colored called serum is squeeze out.
o Serum does not contain fibrinogen. Fibrinogen is changed into fibrin if blood clots.
o Red blood cells has no DNA.
o White Blood Cells has a DNA in nucleus.

II. BLOOD EXAMINATION

BLOOD

o It has been called the circulating tissue of the body.

o Carries oxygen and nutrients to all parts of the body, and carries carbon dioxide and other waste
products back to the lungs, kidneys and liver for disposal.
o For every 600 red blood cells, there are about 40 platelets and one white cell.

ELEMENTS OF BLOODS

1. (45%) formed elements or the solid materials consisting chiefly of cells namely:

a. RED BLOOD CELLS (ERYTHROCYTES)

o The most abundant cells in our blood; they are produced in the bone marrow and contain a protein
called haemoglobin that carries oxygen to our cells. No nucleus.

(NOTE: Hemoglobin picks up and drop-off oxygen)

b. WHITE BLOOD CELLS (Leukocytes)

o They are part of the immune system and destroy infectious agents called pathogens. With Nucleus.

PLASMA (55%)
o This is the yellowish liquid portion of blood that contains electrolytes, nutrients and vitamins,
hormones, clotting factors, and proteins such as antibodies to fight infection.

SERUM
o A straw-yellow colored liquid formed when clotted blood is allowed to stand for some time and the
clot contracts.

PLATELETS (Thrombocytes)
o The clotting factors that are carried in the plasma; they clot together in the process called
coagulation to seal a wound and prevent loss of blood.

FIBRINOGEN
o Is a glycoprotein complex, made in the liver, that circulates in the blood of all vertebrates. Fibrin
clots function primarily to occlude blood vessels to stop bleeding.

HISTORY OF BLOOD GROUPING

 Before the 20th century, physicians have tried to transfuse blood from different blood from one
individual to another. Their attempt often ended in failure because the transfusing blood tended to
coagulate in the body of the recipient causing an instantaneous death.
 (1901) Karl Landsteiner- developed blood typing. This classification also called the A-B-O system.
On 1930, he received a Nobel prize award for his work in blood typing.

 (1937) Rhesus (Rh) Factor- another blood factor is discovered. At present, more than 100 different
blood factors shown to exist.
 Until early 1990’s- forensic scientist uses blood factor technique in linking blood to the person that
originates this blood.
 In theory, no two individuals, except for identifcal twins, could be expected to have the same
combinations of factors.
 IMPORTANCE OF THE STUDY OF BLOOD

1. As circumstantial or corroborative evidence against or in favor of the perpetrator.

2. As evidence in case of disputed parentage.
3. As evidence in the determination of the cause of death and the length of time the survive the
attack.
4. Determination of the direction escape of the victim or the assailant.

BLOOD GROUP
- Differences in human blood are due to the presence or absence of certain protein molecules called
antigens and antibodies.

ANTIGENS (agglutinogen)- located on the surface of the red blood cells. Substances recognized by
the body to produce an antibody to react specifically to it. The most common are A-B-O and Rh system.
ANTIBODIES (agglutinin)- located in the blood plasma. It is a protein that destroys or inactivates a
specific antigen. It ensures that only the blood cells of your blood type exist in the body.

POSSIBLE COMBINATIONS OF BLOOD TYPES

PHENOTYPE- inherited characteristics (blood groups) A,B,AB,O

GENOTYPE- pair of genes represented by AA, AB, BB, BO, AO, OO

IMPORTANCE OF THE APPLICATION OF BLOOD GROUP DATA

- Determination of paternity and maternity
- Determination whether a child born from a married woman could or could have been fathered by
her legal spouse.
- Determination whether a child could or could not belong to a given set of parents in the case of
accidental interchange of infants in hospital.

PLACE OF COLLECTION
1. FLUID BLOOD may be collected from:
o Victims of crime of violence
o Parents and child in case of disputed parentage.

2. Dried Blood or Bloodstain may be collected from;

o Smooth surface like walls, finished floors and table tops/immovable objects
o Hard surface like axe, hammer, knives, stones and crowbars;
o Glazed surface like glass, tiles, and automobiles
o Clothings

FOUR CHRONOLOGICAL TESTS FOR PRELIMINARY TEST

1st (PRESUMPTIVE SCREENING TEST OR COLOR TEST)

o it determines whether the stain contains blood or another substance.
o It determines whether visible stains do or do not contain blood. It is used to demonstrate the
presence of blood.
o Presumptive test produce a color reaction once exposed to specified reagent.

a. Benzidine Test- benzidine solution and hydrogen peroxide (agua oxigenada). It will result to
INTENSE BLUE color.
b. Phenolphthalein Test (Kastle-Meyer) Test- phenolphthalein and reagent and hydrogen
peroxide. It will result to RED PINK color.
c. GUAIACUM TEST (VAN DEEN OR DAY’S OR SCHOINBEIN TEST)- guaiae and hydrogen
peroxide. It will result to BEAUTIFUL BLUE color.
d. LEUCOMALACHITE TEST- leucomalachite green reagent and hydrogen peroxide. It is not as
sensitive than Benzidine Test. MALACHITE GREEN WITH A BLUISH-GREEN OR PEACOCK
BLUE COLOR.
e. LUMINOL TEST- the reaction of luminol with blood results in the production of light rather than
color.
f. TETRAMETHYLBENZIDINE (TMB)- most common test for blood.
g. HEMASTIX TEST- designed as a urine dipstick test for blood. It will result to GREEN color.

2nd- CONFIRMATORY TEST

 o The actual proof that a stain is blood consist in establishing the presence of characteristics
blood pigment, hemoglobin or one of its derivatives.

A. MICROSCOPIC TEST
- useful for the demonstration and mensuration of blood.
- distinction between mammalian and reptilian blood.
- Investigation of menstrual, lochial (blood during birth), and nasal charges.

B. MICROCHEMICAL OR MICROCRYSTALLINE TEST

- HEMATIN CRYSTAL ASSAY (TEICHMAN TEST)

 Using sodium chloride and glacial acetic acid.

 It will result to DARK BROWN RHOMBIC HAEMIN or HAEMATIN CHLORIDE arranged singly or
cluster.

- HEMOCROMAGEN CRYSTAL ASSAY (TAKAYAMA TEST)

 It will result to LARGE RHOMBIC SALMON PINK COLOR arranged in cluster.

- WAGENHAAR TEST OR ACETONE-HAEMIN TEST

 SMALL DARK DIACHRONIC ACICULAR CRYSTALS OF ACETONE HAEMIN.

C. SPECTROSCOPIC EXAMINATION
 the most delicate and reliable test for the determination of the presence of blood in both old and recent
stains.

3RD - PRECIPITIN TEST

o Standard test used to determine whether the stain/blood is of human or animal origin based
on the serum protein.
o Human Blood- white cloudy line or milky precipitate at the contact point of the fluids.

4th – BLOOD GROUPING/TYPING

o In 1900, Karl Landsteiner developed the ABO System of blood grouping.
o These are based on the ability of the blood serum of one person to clump or bring
together the red cells of an individual.
o RBC- antigen or agglutinogen
o Serum- anti-bodies or agglutinins.

BLOODSTAIN PATTERN ANALYSIS

- It is the examination of the shapes, locations and distribution patterns of bloodstains, in
order to provide an interpretation of the physical events which gave rise to their origin.
- Bloodstain and patterns are useful for interpreting and reconstruction of events that
occurred during bleeding.

BLOODSTAIN PATTERN
- A bloodstain pattern is a physical, geometric image created by blood contacting surface, or
by a surface contacting blood.
- The geometric images of interest are primarily those created once blood leaves the body.

TYPES OF BLOODSTAINS DEPEND ON THE FOLLOWING:

 Velocity at which it was travelling;
 Distance travelled;
 Amount of blood;
 Angle of impact; and
 Type of target onto which it lands.

BLOODSTAIN PATTERN ANALYSIS TERMS

1. SPATTER- bloodstains created from the application of force to the area where the blood originated.
2. ORIGIN/SOURCE- the place from where the blood spatter came from or originated.
3. ANGLE OF IMPACT- the angle at which blood droplet strikes a surface.
4. PARENT DROP- droplet from which a satellite spatter originates.
5. SATELLITE SPATTERS- small drops of blood that break of from the parent spatter when the
blood droplet hits the surface.
6. SPINES- the pointed edges of a stain that radiate out from the spatter; can help determine the
direction from which the blood travelled.
7. FORENSIC SEROLOGY- application of the study of blood, semen, saliva, and other body fluids to
legal matters.
 CHANGES IN THE SHAPE OF BLOOD DROPLETS
- Height of the victim
- If the person is walking or running
- The surface textures
- Angle of impact

BLOOD DROPLETS CHARACTERISTICS

- A blood droplet will remain spherical space until it collides with a surface.
- The spherical shape is caused by the surface tension of the blood.

TYPES OF BLOODSTAINS
1. Passive Bloodstain- pattern created from the force of gravity.
2. Single Drop- blood drops that have fallen vertically, whether it be from an injured person or
another object, and landed onto another surface.
3. Transfer Bloodstain- created when a wet, bloody surface comes in contact with secondary
surface.
4. Projected Bloodstain- created when an exposed blood source is subjected to an action or force,
greater than the force of gravity.
a. Arterial Spurt/Gush- bloodstain patterns resulting from blood exiting the body under
pressure from a compromised artery.
b. Cast Off Stains- blood release or thrown from a bloodbearing object in motion. It occurs
when the assailant bloodstained object back before inflicting another blow.
5. Impact Spatter- bloodstain patterns created when a blood source receives a blow or force
resulting in the random dispersion of smaller drops of blood.
a. Low Velocity- gravitational pull up to 5 ft/sec. Relatively larger stains 4mm in size and
greater. Such as blood dropping into blood.
b. Medium Velocity- force or energy in excess of 5ft/sec and reaching no more than 25
ft/sec.
c. High Velocity- Force or energy in excess of 100 feet/sec and greater. Preponderant stain
size of 1mm and smaller mist like appearance.
6. Direction of Movement- the pointed end of the bloodstain faces the direction of travel.
7. Void Pattern- occurs when a person or object blocks the path of the blood. It will help determine
if an object is missing in the scene or if the body is moved.

DROP SHAPE CHARACTERISTICS

1. Target Surface Texture

- Drop on smooth surface- uniform or regular circular shape
- Rough surface- irregular shaped stain with rough or jagged edges.
2. Height
- The higher the droplet falls from, the more blood satellite spatter occurs.
- The velocity of the droplet will be greater, the longer the droplet is airborne.
3. Moving Person
- Droplets dripping from a moving object or person do not drop straight, they will form an
angle.
4. Impact Angle
a. Round- falling straight down at a 90-degree angle.
b. Elliptical- blood droplets elongate as the angle increases from 90 to 0 degrees.
5. Pool Stains
- Refer to the accumulation of blood on a particular surface, generally from prolonged
bleeding from a wound or accumulation of arterial blood.
6. Expiration Stains
- Associated with injury to the respiratory tract, this type of bloodstain is caused by blood
being coughed or otherwise expelled from the mouth.

AREA OF INTERSECTION AND CONVERGENCE

- Location of blood origin can be determined by drawing lines from the different points of
blood droplets to the point of intersection.

WHAT CAN THE INVESTIGATOR EXPECT FROM A BLOOD PATTERN ANALYSIS

o The positions of the victim, assailant, and objects at the scene during the attack.
o The type of weapon that was used to cause the spatter.
o The number of blows, shots, stabs, etc. that occurred.
o The movement and direction of victim and assailant, after bloodshed began.
o It may support or contradict statements given by witnesses.
II. SEMINAL FLUID

SEMEN- a viscid whitish fluid of the male reproductive tract consisting of spermatozoa suspended in
secretion of accessory glands.

Biological Components of Semen

1. Seminal Fluid- has characteristics of alkaline odor, it is viscid, gelatinous and sticky.
2. Spermatozoa or Sperm Cells
- Greek word “Sperma” which means “seed” or latin word “something sown”
- typically has 1.5ml to 3.5 ml is the normal quantity of seminal fluid in single ejaculation with a
usual total number of 400 to 500 million sperm cells. Hypospermia- less than 1.5ml; Hyperspermia-
more than 3.5 ml.

Three Distinct Parts of Spermatozoa

a. Head- is consist of nucleus with densely packed chromosomes covered with acrosomal cap.
b. Middle Piece- consist of mitochondria, located at the short neck which provides energy for
the movement of tail.
c. Tail- responsible for the flagella of the spermatozoa.

3. Flavins- helps to give a yellowish color to semen and caused it to fluoresce under UV Light.

Abnormality in the Production of Spermatozoa

1. Aspermia- a condition wherein males have no spermatozoa at all.
2. Oligospermia- a condition wherein males have abnormally low sperm counts.
3. Necrospermia- refers to a condition in which there are dead or immobilize spermatozoa in the
sperm.

WHERE TO LOOK FOR SEMINAL STAIN?

A. Clothes
B. Body
C. Crime Scene

EXAMINATION OF SEMEN AND SEMINAL STAIN

PHYSICAL EXAMINATION
1. Microscopic Examination-the sperm is stained with and viewed under high power microscope.
The only specific test for semen.

Procedure
a. Transfer a drop of specimen to a glass slide.
b. Add a drop of water or saline solution and cover with glass slip.
c. Examine under the HPO.
d. Observe for the presence of spermatozoa.

Determination of Spermatozoa in Seminal Stain

a. A piece of material is teased on a slide in a drop of water.
b. Allow the smear to dry and then stain with Loffler’s Methylene blue for a minute, wash with water,
dry and examine under the microscope.

Limitation of the Microscopic Test

a. Absence of sperm does not prove that the stain have not been produced by human semen.
b. Elements which may obstruct detection of spermatozoa:
 Nature of Fabric
 Age of Stain
 Condition to which the stain was exposed before reaching the laboratory
 Handling the specimen
c. Some medical jurist believes that these can be no semen without the presence of spermatozoa, but
not true in case of Aspermia and Oligospermia.

2. Ultraviolet Light Examination- seminal stains exhibit bright bluish fluorescence.

3. Semen when dried gives a stiff, starchy feeling to the cloth and produces slight deepening of the
color with the disappearance of the odor.
4. Have appearance or outline of contour map.

CHEMICAL EXAMINATION
1. Florence Test- developed by Dr. Florence Lysons, based on the formation of crystals of choline
periodide.
 - It will result to a Crytals of Choline Periodide,which are DARK BROWN, RHOMBIC OR
NEEDLE-SHAPED as seen under the microscope.

Procedure
a. Cut a portion of the stain and divide into small bits then soak in saline solution.
b. Transfer into a slide, tease and evaporate the liquid.
c. Add a drop of Florence reagent and cover with cover slip.
d. Examine under microscope.

Limitation of the test

a. Clothes with seminal stains that are not dried thoroughly will make Choline Periodide to
decomposed easily.
b. If stain is wet and mixed with blood.
c. After 24 hours, it will yield negative result due to rapid decomposition but still spermatozoa can be
detected.
d. Even after a long period of 2 ½ years, it will still give positive result.

2. Barberio’s Test- crystals which are SLENDER YELLOW TINTED RHOMBOID NEEDLE SHAPE
of spermine picrate. Reagent use is saturated aqueous or Alcoholic Solution of Picric Acid.

Procedure
a. Soak a piece of stained material in a 2.5% solution of Trichloroacetic Acid for one hour in a test
tube.
b. Centrifuge the test tube.
c. Get the clear liquid part and add to an equal amount of a saturated Aqueous or Alcoholic Solution
of Picric Acid on a glass slide.
d. Observe under a microscope.

3. Acid Phosphatase Enzyme Test- it is used in high levels of fresh seminal stains and yield rapid
result. This test is based fundamentally upon the considerable high acid phosphatase content of
human male ejaculate. It uses a reagent of 100 mg of Alpha napthyl phosphoric acid and 200 mg
of Brentamine fast blue B.

Procedure
a. Moisten a piece of filter paper with water.
b. Swab the stained area with the filter paper.
c. The acid phosphatase will be transferred to the filter paper.
d. Add a drop or two of Alpha naphthyl phosphoric acid and Brentamine Fast Blue B.

- it will result to a PURPLE COLOR indicative of the presence of acid phosphatase.

4. Alternative Acid-Phosphatase Test- Alphanaphtol by the acid phosphatase combines with the
diazonium salt to form the color. The reaction takes place within 30 seconds on fresh stains.
- It will result to Orange-Red pigment.

BIOLOGICAL EXAMINATION
- test proposed by Farnum in 1901. Also known as spermato-precipitin to determine whether
semen comes from animal or human.

OTHER STAINS OF MEDICO-LEGAL INTEREST

1. Obstetrical and Gynecological Stains- refers to the examination at the scene of the crime in
cases of criminal abortion, infanticide, and sex offenses.
2. Excrements- this refers to waste matter discharged from the body especially feces.
Adults- yellowish brown
Infant- Greenish Yellow
3. Paints- this refers commonly to as pigmented coating. Paint can reveal vital evidence for the
forensic scientist. Some examples of transfer are:
a. Paint recovered from the clothing of a road traffic accident victim.
b. Paint from two or more vehicles involved in a collision.
c. Paint recovered from a tool that may have been used in housekeeping with paint from the
point of entry.
4. Rust Stains- this resembles bloodstains, reddish-brown in color, insoluble in water

COLLECTION, PRESERVATION, PACKING AND TRANSIT OF SEMEN STAINED SPECIMENS

 1. Seizure of wearing apparel must be done as soon as possible. It often happened that washing
clothes and undies has destroyed important traces.
2. During packaging, ensure that the apparel and/or fabric shall not make any friction between the
apparel and the stain. Dried semen is very brittle and may break into small pieces if not handled
properly.
3. Specimen should not be rolled for transit. Gently lay between two sheets of cardboard or similar
material which are tied together with a string to avoid friction.
4. Smaller objects like hair should be placed in a test tube or corked.
5. Specimen must be thoroughly dried before packing.
6. Fluid semen should be placed in a test tube. It may be preserved by a few drops of 10% solution of
formalin during hot weather where there is danger of putrefaction.

III. SALIVA

SALIVA
- it is the secretion of the mouth that is important in digestion and comprise of cells and
secretions from the salivary and parotid glands
- Humans produce 1-1.5L of saliva a day.

HOW IS SALIVA DETECTED?

1. Enzyme that breaks down starch
2. Found in many body fluids

TEST FOR PRESENCE OF SALIVA

1. STARCH-IODINE TEST
- Iodine is used to test for the presence of starch.
- The amylase in starch reacts strongly with iodine and form a DARK BLUE COMPLEX, while
amylopectin develops a REDDISH-PURPLE COLOR.

2. DNA IN SALIVA

IV. FIBER ANALYSIS

HAIR
- Is the specialized outgrowth of the skin which occur everywhere on the human body except
on the palm of the hand and sole of the foot.
- It grows at the rate of 0.3 to 0.5 mm per day.
- Hairs ranges from scalp hair, pubis hair, auxiliary hair.
-
HISTORY OF HAIR EXAMINATION

1847- First used as physical evidence

1897- Rudolph Virchow become the first person to do an in-depth study of hair.
1906- Hugo Marx wrote a paper on the use of hair in forensic investigation to determine
identity.
1931- Dr. Paul Kirk works on new ways to improve the use of hair in forensic investigations.

A. Collection, Packing and Preservation of Hair

COLLECTION OF SPECIMEN
1. Complete search at the crime scene must be done. Use vacuum cleaner.
2. All hair in the questioned specimens should be submitted but do not mix hairs at different
places.
3. Search for and collection of hair evidence should begin as soon as possible. Hair evidence is
easily transferred to and from the crime scene
4. Collection should be done by:
a. Hand- if the location of the hair is important.
b. Lint Rollers
c. Special Filtered Vacuum Cleaner- collect hairs and fibers in mass from carpet, bedding, etc.
5. In vicious assault and murder cases, obtain the clothing of the victim from the hospital or
morgue to avoid lost of evidence by careless handling and to prevent the clothing from being
destroyed.
6. Representative samples of hair from the victim as well as the suspect should be obtained if
possible. Root must be intact.
7. Don’t mix known samples of hair from different parts of the body.
8. Hair evidence should be looked for in the following: clothing, combs, weapons, pockets, fingers,
hats, and etc.
9. Get samples from both victim and suspect.
10.Best way to collect hair is by combing.
 PACKING OF SPECIMEN
1. Hair evidence should be packaged into paper packets.
2. The hairs should be placed in a folded paper or in a white mailing envelope, but the corners of
the envelope should be sealed with a scotch tape.
3. Do not secure the hair samples to a piece of paper with scotch tape as this will damage the hair.
4. All foreign fibrous debris should be removed from the submitted specimen.
5. Fragmentary hairs or underdeveloped hairs are not suitable for examination.
6. Areas on an object containing hairs should be protected with cellophane or paper tape over the
area before wrapping the object from transmittal to laboratory.

PRESERVATION OF SPECIMEN
1. Place in a pill box or test tube.
2. Properly folded, sealed and labeled.

TWO KINDS OF HAIR

 Real Hair- generally long and stiff.
 Fuzz Hair- generally short

PARTS OF THE HAIR

 Root- portion embedded in the skin
 Shaft- portion above the surface of the skin. Most distinctive part of the skin.
 Tip- sometimes termed as point. The distal end of an uncut hair shaft.

Parts of the Shaft

 Cuticle- outer layer of the hair shaft
 Cortex- tells the race of the hair whether Negroid, Caucasian or Mongoloid.
 Medulla or Core- tells whether the hair belongs to animal or human

HUMAN VS ANIMAL HAIR

1. Medulla in a human is smaller (medullary index of less than one-third); and medulla in animals is
very thick (medullary index of one-half or greater)
2. Cuticle in humans is imbricated; and the cuticle in animals is coronal or spinous
3. Pigmentation in animal hair is denser than human hair.
4. Animal hair can change colors in banded patterns; human hair cannot.

TEXTILE FIBERS
- Derived from the Latin Word “Textilis” and the French word “Texere”. Means to weave.

CLASSIFICATIONS OF TEXTILE FIBERS

a. Natural Fibers
 Vegetable Fibers
 Animal Fibers
 Mineral Fibers
b. Synthetic Fibers or Artificial (Man-Made)

Test for Textile Fibers

a. Burning or Ignition test
- a test that determines whether fiber is mineral, animal or vegetable fibers
b. Fluorescence Test
- Vegetable fibers exhibit a yellow fluorescence in ultraviolet light whereas animal fibers show
bluish fluorescence.
c. Microscopic Examination
- The most reliable and best means of identifying fibeer.
d. Chemical Analysis of Fibers
 Staining Test- only animal fibers react with picric acid while vegetable fiber reacts with
stannic chloride.
 Dissolution Test- wool and silk dissolved in Sodium Hydrochloride while cotton, linen, wild
silk and cellulosic silk do not.

GLASS
- Is a supercooled liquid which possesses high viscosity and rigidity.

COMPOSITION OF GLASS
- Silica (SiO2), boric oxide (B2O3) and phosphorus pentoxide (P2O5).
 TYPES OF GLASS FRACTURE
1. Radial Fracture- primarily fracture resembles the spokes of a wheel where the radiating
rod originates at a common point.
2. Concentric Fracture- secondary fracture having the appearance of circles around the
point of impact connecting one radiating crack to the other, thus forming triangular pieces
of glasses.

SIGNIFICANCE OF DETERMINING THE TYPE OF GLASS FRACTURE

1. Point of Impact- the front of the glass can be determined due to the accumulation of dust and
dirt on the glass.
2. Direction of Impact- a bullet will make a clear-cut hole in the side of the entrance rather than on
the exit side. If a shot is fired perpendicularly, it will give a crater of uniform flaking. If the shot is
fired at an angle from the right, the left exit side of the glass will produce more flaking and vice
versa.
3. Cause of Fracture
B. Due to Natural Means- exhibits a plain wavy lines.
C. Due to Mechanical Means- exhibits a regular pattern of radial/concentric
fracture.

FACTORS TO BE DETERMINED IN GLASS FRACTURE

a. Point of Impact
b. Position of the Shooter
1. Perpendicular Shot- exhibits an even distribution of chipping on the exit side of the glass.
2. Angle from the Right- heavy flakings or chippings on the left side of the glass.
3. Angle from the Left- heavy flakings on the right side of the glass.

AGE OF FRACTURE
1. Fresh Fracture- exhibits a regular pattern of radial/concentric fracture.
2. Old Fracture- presence of a short extension lines at the end of the radial fracture.

PART II: FORENSIC TOXICOLOGY

TOXICOLOGY
o Branch of science that defines the nature, effects, and detection of substance that is present to a
specific matter.

CLINICAL TOXICOLOGY
o Deals with human diseases caused by, or associated with abnormal exposure to chemical
substances.

FORENSIC TOXICOLOGY
o From the Greek words, toxicos and logos.
o Study of the symptoms, mechanisms, treatments and detection of poisoning.
o It centers on the determination of toxic substances in the human tissues, organs and body fluids
such as urine and blood, and the subsequent determination of the cause of death due to toxins.

IMPORTANCE OF TOXICOLOGY
1. To verify if the cause of death is poisoning.
2. To be able to treat as the occasion demands.
3. To forward justice.

TOXIN
- A harmful compound that is usually produced by living cells or organisms and can cause disease or
harm when introduced into the body’s tissue.

TOXICANT
- are synthesized chemical substances that impact biological functions in other organisms.

POISON
- A toxin that enters the body by being swallowed, inhaled, or absorbed through the skin.
- “All substances are poisons. There is none which is not a poison. The right dose differentiates a
poison and a remedy”- Paracelsus (1532)

TYPES OF POISONING
1. Hyperacute Poisoning- produced by a single massive dose. Death occurs very rapidly without
showing any signs and symptoms.
2. Acute Poisoning- one in which there is prompt and marked disturbance of function or death within a
shorter period.
3. Subacute Poisoning- gradually over some time. Doses taken are small.
 4. Chronic Poisoning- a kind of poisoning in which there is gradual deterioration of functions of tissues
and may or may not result in death.

VENOM
- A toxin that enters the bloodstream through injection or an injury.

TOXINS ARE CLASSIFIED AS:

A. Based on Effect:
1. Exotoxins- toxins which are excreted by organisms.
2. Endotoxins- which are produced when bacteria are lysed.
3. Hemotoxin- toxins that destroy red blood cells.
4. Phototoxin- causes one to become photosensitive.
5. Biotoxins- toxins that are biological in nature.

B. Based on Origin
1. Animal/Toxin- a poison produced by living organism stimulating antibodies.
2. Vegetable
3. Mineral
4. Microbial
5. Synthetic- manufactured by chemist such as drug.

C. According to Chemical Properties

1. Volatile Poisons- poisons that easily changes into gas. poisonous compounds that can be isolated
using steam distillation and analyzed using Gas Chromatography.
2. Non-Volatile Substances- those poisons that do not easily turn into gas.
3. Anion- toxic effect through the negative ion effect of certain substances such as salt.
4. Metallic
5. Miscellaneous- ex. Pesticides

D. According to Action (Physiological)

1. Irritants- by direct contact, this poison inflames the mucous membrane or the parts it comes in
contact resulting to nausea, vomiting, pain, and diarrhea
o Inorganic- Non-metallic: Phosphorus, halogens, arsenic, mercury.
o Organic- vegetable poisons such as castor oil, cotton oil, and aloes. and Animal Poisons such
as snakes, insects and others.
o Mechanical- powdered glass, chopped hair, diamond and wood dust.
2. Corrosives- by direct, chemically produces local destruction of tissues.
o Strong Acid- sulphuric, nitric, hydrochloric
o Strong Alkalies- hydrates and coarbonates of sodium, potassium and ammonia.
3. Neurotics- those that effect the Central Nervous System (CNS):
 Cerebral Neurotics (Narcotics)- inducing drowsiness, sleep stupor, complete or
incomplete insensibility or loss of feeling.
 Spiral Neurotics (Tetanics)- a poison that act on the spinal cord producing spasmodic or
continuous contractions of muscles resulting in stiffness of the parts to which they are
attached. Such as Strychnine.
 Cerebrospinal Neurotics
o Delirants- are poison that act on the brain causing disorder of mental functions
resulting to confusion of will.
o Depressants- a substance that retards the physiological action of an organ.
o Aesthetics/Exhaustive- poisons that cause marked loss of vital and muscular
power or general weakness.
4. Cardiac- nicotine
5. Asphyxiac
o Irrespirable Gas
o Cyanides

6. Cumulative Poison- is one that increases suddenly in its intensity of action after gradual additions of
it.

E. Actions of Poison
1. Local- the changes or disturbance produced on the part with which the poison come in contact.
2. Remote- the changes or disturbance produced in distant parts away from the site of application.
3. Combined- the effect of the poison is not only localize at the site but also affects remote organs.

METHODS OF EXAMINATION
1. ISOLATION- when the submitted specimen is in pure form, the poison must be first isolated.

2. IDENTIFICATION- the method employed for the identification of poison is specific