FORENSIC CHEMISTRY

Lecture Module

I.J, Buan | B.M.Hermano | K.D.G. Martin

M.A.Rivera | E.A.T Romero | K.G.V.Padilla

This module is a property of NEUST

 TABLE OF CONTENTS

Unit I. Introduction to Forensic Chemistry 1

Unit II. The Crime Scene 5

Unit III. Forensic Toxicology: Poison 10

Unit IV. Classification of Drugs 19

Unit V. DNA 26

Unit VI. Blood 31

Unit VII. Semen 41

Unit VIII. Hair and fibers 44

Unit IX. Gunpowder and Explosives 52

Unit X. Chemistry of Combustion and Arson 59

Preface

The lecture module in Forensic Chemistry is intended for 3rd-year

students who are taking BS Criminology for the A.Y 2020-2021 at Nueva Ecija
University of Science and Technology. This lecture module aims to demonstrate
the application of chemistry in the identification of physical evidence. It includes
the collection, preservation, examination, and study of blood, semen, and other
body fluids for DNA, gunpowder, and explosives, hair and textile fibers as applied
to crime investigation.
The authors hope that the students who will be using this lecture module find it
useful in their studies as they prepare to become the future criminologists of the
country.
UNIT I. INTRODUCTION TO FORENSIC CHEMISTRY
Overview
“If the law has made you a witness, remain a man of Science. You have no
victim to avenge, no guilty or innocent person to ruin or save. You must bear
witness within the limits of Science” --Brouarded. Forensic chemistry focuses on
the application of chemistry in the identification of physical evidence that uses in
crime investigation.
Learning Objectives
At the end of the unit I am able to:
1. Define forensic science and forensic chemistry;
2. Relate the philosophy of forensic science to the investigation;
3. Apply scientific methods in forensics;
4. Identify the scope of forensic chemistry;
5. Identify the stages of forensic chemistry concerning the rules in the practice
of Forensic Chemistry.
Setting up
Directions: Read the situation below. Answer the question briefly.

In 1996, Nickelodeon introduced us to “Blue’s Clues,” an animated preschool

television series. It follows the adventures of Blue, a blue-spotted puppy, her
owner, Steve, and Blue's friends. Blue leaves paw marks to the things and Steve
should find it and relate each thing at the end of the show to know what Blue
wants to express. Steve is trying to act as a detective during the show.
Question: How Steve gathers clues and connect each clue to solve the problem.
What is Steve’s characteristics you notice similar to a crime investigator?

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Lesson Proper
What is Forensic Science?
Forensic science is often shortened to forensics. The application of a broad
spectrum of sciences to answer questions of interest to a legal system. The word
forensic comes from the Latin adjective forensic, meaning "of or before the forum.”
Forensic Science is any science used for the law and therefore provides
impartial scientific evidence for use in the courts of law. The multidisciplinary
subject, drawing principally from chemistry and biology, but also physics, geology,
psychology, social science, etc.
What is Forensic Investigation?
Forensic Investigation is a combination of forensic science, law, and criminal
investigation and can be described as a problem-solving activity.
The interest of Forensic Investigation
1. Observe physical evidence
2. Determine identity
3. Determine origin
Philosophy of Forensics
The key to criminalistics is the ability to think clearly when faced with the
daunting confusion of the average crime scene. One must rely on experience,
intuition, and abductive reasoning (testing a likely scenario against the facts.
Scientific Methods in Forensics
1. Observe problem or questioned evidence and collected data.
2. Make a hypothesis or possible solution to the problem based on the
observations, given a directed work plan.
3. Examine, test, and analyze to support or refute the hypothesis.
4. Use deductive reasoning to make a determination as to the significance of the
evidence.
5. Evaluate and verify all evidence and determine a conclusion.
Forensic Chemistry Defined as
Forensic Chemistry – that branch of chemistry that deals with the application of
chemical principles in the solution of problems that arise in connection with the

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administration of justices. It is chemistry applied in the elucidation of legal
problems.
Scope of Forensic Chemistry
1. It includes the chemical side of a criminal investigation.
2. It includes the analysis of any material the quality of which may give rise to
legal proceedings.
3. It is not limited to purely chemical questions involved in legal proceedings.
4. It has invaded other branches of forensic sciences notably legal medicine,
ballistics, questioned documents, dactyloscopy, and photography.
Four Stages in the Practice of Forensic Chemistry
1. Collection or reception of specimen to be examined;
a. Sufficiency of samples
b. The Standard for comparison
c. Maintenance of individuality
d. Labeling and sealing
2. The actual examination;
3. The communication of results of examination; and
4. Court appearance.
Six Golden Rules in the Practice of Forensic Chemistry
1. Go slowly
2. Be thorough
3. Take notes
4. Consult others
5. Use Imagination
6. Avoid complicated theories
References
PSUPT. Jaime Rodrigo L Leal, MD, Forensic Science, [Powerpoint slides].
Melcon S. Lapina, MSCrim, Forensic Chemistry, and Toxicology, [Powerpoint
slides]

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Assessing Learning
Activity 1.

Name:_______________________________ Score:_________

Course/Year/Section:___________________ Date:__________

Directions: Essay. Analyze the criminal case below. Answer the question after the
paragraph.
“There have been a series of cases where doubts were raised as to whether the
dangerous drugs tested actually. In People v. Dahil and Castro (2015), the forensic
chemist who examined the suspected substances certified that she had “no personal
knowledge as from whom and where said the substance was taken.” In People v.
Beran, cited in People v. Dahil (2015), the police investigator claimed to have
personally delivered the drugs to the laboratory for testing, “but there was no
showing who received the drug from him.” - People v. Dahil and Castro. 12 Jan 2015.
G.R. No. 212196.
This shows lapses in the investigation of illegal drug cases in the Philippines.
If you were the police investigator in the case, What should have you done
differently. Relate your answer in the “Four Stages in Practice of Forensic
Chemistry”.

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UNIT II. THE CRIME SCENE

Learning Objectives

At the end of this activity I am able to:

1. Describe the basic processes applied in processing crime scenes.

2. Be familiar with the basics of evidence handling

During forensic investigations, if the evidence is mishandled or is lost at the scene, no

amount of laboratory analysis can reverse this problem and usually, the scene cannot be
revisited to attempt at obtaining additional evidence. Personnel who are responsible for
examining the scene of any crime can include a police officer, a detective, a crime scene
examiner, or a forensic scientist. At present, with the rapidly increasing technology used
to examine crime scenes, there is a need for those involved in dealing with a crime scene
to be trained. The solution to many crimes depends on a thorough investigation of the
crime scene conducted by specialist examiners and by other experts.

Many types of evidence can be collected in a scene. These are handled by different
specialized departments such as:

• Fingerprint – recover and photograph fingermarks at the crime scene for

comparison and identification
• Scene of Crime – assess, control, record the scene by document and photography,
examine and recover all types of evidence, interpret the scene for evidence, collect
and control exhibits, coordinate and prepare reports and statements of evidence
• Photographic Services – produce photographic and video images when non-
routine techniques are required, used in areas such as traffic, surveillance and
public order offenses
• In-Force Laboratories and Scientific Services
Important Considerations in a Crime Scene

• At the crime scene, the main task of an officer is to preserve the scene. Restrict
access to any area that might have evidence and protect all evidence.
• Prevent contamination of any part of the scene by others. A written record of all
persons attending the scene must be available and maintain a communication
link.
• Protective clothing must be worn.
• Look for signs of disturbance or struggle.
• External (outdoor) locations require fast and decisive actions to control and
preserve evidence due to weather conditions.
• The priority is to examine and recover a body of the victim(s) in a controlled
manner with the minimum evidence disturbance.

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 • At all stages, a photographic record is made.
• Leocard’s Principle- evidence from the attacker is most likely to be on the body
itself, be cautious as hasty movements will disturb this evidence.
• Surfaces will be searched, will be taped to recover any contact traces.
• The victim is regarded as a scene. A post-mortem will have to be performed. Take
all relevant samples.
• The selection of best evidence types fit the needs of the case. Evidence recovery
must be completed in full. Careful selection of evidence and samples can limit the
workload of the forensic analysts.
• When evidence is first taken, it is sealed in a secure container and an exhibit label
is attached. It contains details of what the item is, where it was found, and when.
A reference number is given to an exhibit based on the initials of the person taking
it. Indicate every person who has responsibility at any stage for that particular
exhibit, they must sign and date the label in sequence. Secure lid with adhesive
tape to complete seal.
• More sensitive DNA analysis is now available.
The Basics of Handling of Evidence

• Only a few kinds of physical evidence can be individualized. Individualization

means that an item of evidence comes from a unique source. It can be shown to
be directly associated with a specific individual source (fingerprints, tool marks,
and fired bullets).
• The majority of physical evidence found at crime scenes can only be identified.
Identification means that the items share a common source.
• Most physical evidence cannot definitely connect a suspect to crime as fingerprint
evidence can. This should not diminish the usefulness of that evidence. Physical
evidence that is identified can corroborate testimony, place a subject at a scene,
etc.
• Before any evidence is seized, the need for a search warrant or court order should
be considered. In some cases, the physical evidence seized at a crime scene may
be inadmissible in court for lack of a warrant.
• To prove that the integrity of the physical evidence has been maintained, a chain
of custody must be demonstrated.
• Evidence must be held in a secured area prior to transportation to court. Evidence
reasonably assumed to have been tampered with by unauthorized persons
because it was kept in an unsecured area may be inadmissible in court. Evidence
should be maintained in a specific secured area, with limited access by authorized
persons.
• Too much handling may obliterate fingerprints, dislodge minute trace evidence
such as hair, fibers, and debris, break apart brittle evidence, or contaminate
evidence. Forceps, latex gloves, and special containers may be necessary for
handling physical evidence.
• Microscopic or trace evidence presents unique collection problems. A number of
techniques are available to collect this type of evidence. Vacuum cleaners

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 specially equipped with traps can be used to collect trace evidence such as hairs,
fibers, glass fragments, and debris.

Fig, 2. A Typical label used to identify an exhibit and crime scene examiners at work.

References

Fisher, B. A., & Fisher, D. R. (2012). Techniques of crime scene investigation. crc Press.
Khan, J. I., Kennedy, T. J., & Christian Jr, D. R. (2011). Basic principles of forensic chemistry.
Springer Science & Business Media.

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Assessing Learning

Activity 2
Name:_______________________________ Score:_________
Course/Year/Section:___________________ Date:__________
Sample Case

The victim of this crime had lived in a semi-detached house on a small estate in the town.
When the victim returned to her house, she was surprised by an intruder. The intruder
subsequently put her in his vehicle and drove to a secluded lay-by on the main road out of
the town. He removed her from the vehicle, opened a gate to a field, carried her across to the
far side, and left her under a hedge. Her handbag and some items of female clothing were
thrown over the hedge into the field beyond. The following morning, a farmer out shooting
rabbits found the partially clothed body under the hedge and called the Police. When the
victim did not keep an appointment with a male friend that evening, he telephoned the
house. There was no reply. The following morning, when he was still getting no reply to his
calls, he became anxious and went round to the house. He saw that a window had been
broken and was open. He called the police.

• Note that the first person(s) at any scene must cause some disturbance.
• As an analyst, look at the scene based on knowledge and experience as the facts
fall into place.
• There will be pressure to deal with the crime scenes quickly since these may
provide evidence of immediate value in locating or identifying the murderer(s).
At the start of the inquiry the following questions are posed:

• Where, when, how did the victim die?

• How was the victim attacked?
• What does the evidence tell us?
• Was the victim attacked in more than one location?
• Did the victim know the attacker or was he a stranger?

The police officers assigned to this case faced these problems in terms of the crime
scene”
(a) The victim's house may have been broken into and contain evidence of an assault
leading to murder and traces from the intruders.
(b) The field area surrounding the body may have evidence from the murder(s), from
the vehicle used as well as items from the victim.

(c) The body of the victim may have evidence of the cause of death, nature of the
assault, and traces of contact between the victim and her attackers. The victim has
control samples.

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 (d) The murderer may have returned to the crime scene to take evidence of the crime
and eliminate traces.
(e) The vehicle of the murderer(s) may have evidence from the victim and evidence
that an assault occurred in the vehicle.
I. As the First Officer Attending assigned at the case mentioned, what are your
primary responsibilities? What is the proper way to restrict access to the crime
scene?

II. Define the “common approach path” in dealing with the bodies of victims.

III. What is a post-mortem exam? Who is authorized to conduct one?

IV. Enumerate the sources of biological samples that must be taken during a post-
mortem exam.

V. If you are the detective assigned to this case, enumerate possible evidence that you
must look for to answer the problems stated above.

VI. What will be the next phase of scene examination after a suspect has been identified?

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UNIT III: FORENSIC TOXICOLOGY: POISON
Learning Objectives:
After reading this unit, you should be able to:
1. describe the main groups of poisons;
2. discuss the main types of commonly abused drugs, with particular reference to
their chemical nature, physical forms and effects;
3. explain the different factors that influence the toxicity of a substance;
4. appreciate the different routes of uptake of toxic compounds into the human body
and the means by which they are subsequently eliminated; review the
information sought during the analysis of samples for drugs and other poisons
and recognize the means by which such analyses may be carried out; and
5. know the elimination procedures applied in case of absorbed and unabsorbed
poisons as well as the various types of antidotes and their importance.

Forensic toxicology is the scientific study of poisons in relation to the law. A

poison is any substance that produces a detrimental or lethal effect when introduced into
or otherwise taken up by an organism. Drugs of abuse are those taken by individuals for
the mood and mind-altering effects that they induce.

Classification of poisons can be done according to:

A. Mode of Action. It is a functional or anatomical alteration at the cellular level,
stopping from the exposure of a living organism to a material or a substance.
1. Corrosive Poisons. It is simply a highly active irritant and not only produces
inflammation but also actual ulceration of the tissues. Chiefly, it fixes, abolishes, and
corrodes the surface with which it comes in contact, they produce local action. This group
comprises strong acids, such as sulphuric acid, nitric acid, hydrochloric acid, organic
acids, such as oxalic acid, carbolic acid, acetic acid, salicylic acid; and concentrated alkalis
such as caustic soda, carbonates, ammonium, sodium, and potassium.
2. Irritant Poisons. It causes pain in the abdomen, vomiting, and purging. In post-
mortem appearance, they are usually evident to the naked eye and show redness or
ulceration of the gastrointestinal tract. This group is further divided into inorganic,
organic, and mechanical substances. Corrosives in dilute solutions act as irritants. (i)
Inorganic irritant poisons consist of non-metallic and metallic poisons. (ii) Organic irritant
poisons comprise of animal and plant poisons. (iii) Mechanical substances include coarsely
powdered glass, chopped hairs, dried sponge, and diamond dust.
3. Systematic poisons. These
directly affect the main organs of
the body system. It includes the
nervous system (neurotics),
cardiovascular system (cardiac),
respiratory system (asphyxiate).
(i) Neurotics poisons act chiefly
on the nervous system through
some neurotics have a local irritant action. All alkaloids poisons fall into this group. (ii)
Cardiac poisons acting on the heart and it includes digitalis, oleander, aconite, and

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nicotine. (iii) Asphyxiant poisons act on the respiratory system and it includes irrespirable
gases such as carbon monoxide, carbon dioxide, sewer gases, and some war gases.
4. Miscellaneous poisons. These have different pharmacological actions are put
together in this group. It includes analgesics, antipyretics, antihistaminic, tranquilizers,
antidepressants, street drugs, and designer drugs.
B. Nature of Poison
1. Gaseous Poisons. If inhaled, hamper the competence of the blood as a carrier of
oxygen and may damage the tissues of the air passages and lungs such as carbon
monoxide, carbon dioxide, chlorine, nitrous oxide, and tear gas.
2. Volatile inorganic poisons. Acute poisoning with volatile substances usually
follows the slow inhalation of vapors in order to become intoxicated. Cyanide, phosphine,
arsine, phosgene, and chloride are few examples.
3. Volatile organic poisons have a high vapor pressure at ordinary room temperature
such as ethanol, formaldehyde, and acetaldehyde.
4. Non-volatile inorganic (anions) poisons. An anion is a negative ion. Many
compounds used in the home and/or workplace contain toxic anions, for example, some
weed killers, bleaching agents, and insecticides.
5. Non-volatile inorganic (cations) poisons. Examples are mercury, arsenic, barium,
thallium, lead, antimony, and bismuth.
6. Non-volatile organic neutral poisons such as pesticides, any chemical substance
that kills organisms regarded as pests. Examples are organophosphates, organochlorines,
carbamates, and paraquat.
7. Non-volatile organic acidic compound (acidic drugs). These drugs readily react
with bases to form salts. A few of the examples are barbiturates, phenolic compounds
(phenol, cresols), and salicylates.
8. Non-volatile organic alkaline
compounds (basic drugs). If the drugs
contain a nitrogen atom with a lone pair
of electrons available for reaction with
protons they will behave as bases only.
Examples alkaloids and
benzodiazepine.
9. A toxin is any poisonous
substance that is naturally produced by
an organism. They may be taken
accidentally but have also been used as the means of suicide or as an agent of murder.
Classification of Poisoning
1. Fulminating poisoning is produced by an enormous dose. In fulminating
poisoning, death occurs rapidly, sometimes without preceding symptoms and the patient
appearing to collapse suddenly.
2. Chronic poisoning. A small amount of poison is repetitively administered for a
long period produces toxic effects. The substances commonly causing chronic poisoning
are metallic poisons, opium, antimony, and arsenic.
3. Sub-acute poisoning shows features of both, acute as well as chronic poisoning. It
generally refers to continuous or repeated exposure to a chemical for more than 72 hours
but usually less than 1 month.

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 4. Acute poisoning is caused by an excessive single dose, or several doses of a poison
taken over a short interval of time, whatever amount of poison is taken and the toxicity
is manifested immediately.

Types of Poisoning
1. Homicidal poisoning is the administration of poisonous substance by a person to
kill another. One of the most demanding poisons of such nature is arsenic.
2. Suicidal poisoning is the ingestion of some poisonous substance by the individual
himself for self-killing. This includes opium, barbiturate, organophosphorus, carbolic
acid, and copper sulfate.
3. Accidental poisoning takes place due to the storage of poisonous and non-
poisonous substances at the same place or other incidents like the snake’s bite.
a. Food poisoning is widespread in third world countries, mainly due to a relative
lack of sanitation and public hygiene. While, the biological causes of food poisoning are
microbes, parasites, fungi, plants, animals, the chemical cause cannot be ignored such as
chemical contaminants (heavy metals, and pesticides) and food additives.
b. Iatrogenic poisoning arises from the action of a doctor said of any adversative
condition in a patient resultant from treatment by a physician or surgeon, for instance,
death after injection of a wrong solution or of an appropriate solution in an inappropriate
manner.
c. A synergistic effect occurs when the collective effects of two chemicals are much
greater than the sum of the effects of each agent given alone. For example, both carbon
tetrachloride and ethanol are hepatotoxic compounds, but together they produce much
more liver injury.
d. Agro-chemical poisoning is due to other pesticides that are relatively uncommon
except among individuals who are occupationally exposed. The commonest agents are
organophosphates, carbamates, chlorinated hydrocarbons, pyrethroids, and aluminum.
e. Drug poisoning. Drug abuse refers to the use usually by self-administration of any
drug in a manner that deviates from the approved medical or social pattern within a given
culture. The overdosage of the drug commonly occurs accidentally, intravenous injection
carries the highest risk of accidental overdosage, and when it occurs, the drug cannot be
retrieved.
4. Miscellaneous poisoning a Judicial poisoning or the administration of the lethal
injection to the condemned convict.
a. Abortifacient poisoning. Abortion is the termination of pregnancy by eliminating
a fetus or embryo from the womb before it can survive on its own and an abortifacient is
a substance that induces abortion.
b. Stupefacient poisoning. The term “stupefaction” is loosely applied to the process
of representing a victim suddenly debilitated by exposing him to a deliriant poisons such
as Datura, in order to facilitate robbery or rape. Even cigarettes may be adulterated in a
similar fashion.

Factors affecting toxicity

▪ Paracelsus (1493–1541) stated that ‘All substances are poisons; there is none that
is not a poison. The right dose differentiates a poison from a remedy.’
▪ The level of the dose required to provoke a reaction in an organism is reliant on
the toxic properties of the substance in question.

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 ▪ Plotting the percentage response of a group of
animals (or cells) against the log of the dosage (the dose per
unit weight or surface area of the target organism)
produces a typical S-shaped (or sigmoid) curve or the dose-
response curve and can be used to determine the LD50,
defined as the dose at which 50 percent of the test
organisms die. This is used to compare the relative toxicity
of different substances and the results from toxicity testing
on laboratory animals may be used to determine the effects
on humans.
▪ It is significant to understand that the poisonousness of a material is determined
not only by its characteristic toxic properties but also by a number of factors relating to
the individual exposed to it. Also, previous exposure to a particular substance can have a
significant effect on how an individual reacts to the next dose. This may lead to one of the
three following scenarios:
▪ Sensitization. It is characterized by an enhanced immune response like for
example, a previous contact to a particular substance may lead to the development of
sensitization when an individual encounters it for a second time.
▪ Tolerance. This condition may progress when an individual is repetitively
exposed to a particular substance. Thus, the dose must be either increased or given more
frequently in order to have the same effect as when it was first administered. Drugs such
as amphetamines, barbiturates, benzodiazepines, and opiates (like heroin and morphine)
can all lead to the development of tolerance.
▪ Accumulation. The time is taken for the concentration or amount of poison in the
body (or a given part of the body) to halve is called the half-life, t1/2. Some poisons, known
as cumulative poisons, have values of t1/2 that are long enough to allow chronic exposure
to sublethal doses to lead to the accumulation of the poison within the body. Heavy metals
such as lead and mercury, and the metalloid arsenic, are well-known cumulative poisons.

Routes of uptake and elimination of drugs and other toxic substances

The lethal influence of a possibly toxic substance is not used until that material
comes across a biological system. Such substances may pass in the body by one or more
of the routes of uptake outlined below:
▪ Ingestion. In this means of uptake, potentially toxic materials are taken into the
gastrointestinal tract through the mouth. Examples are drugs that administered orally
(abuse or therapeutic use) and toxins present in foodstuffs.
▪ Inhalation. Intake of gases, vapors, and/or particles into the lungs. This may result
from accidental exposure, for example to carbon monoxide in the ambient environment,
or deliberate activity like by the abuse of solvents.
▪ Skin contact. The skin forms an effective barrier against many potentially toxic
materials. However, some poisonous substances, for example, phenol and organic
mercury compounds, are capable of penetrating this barrier.
▪ Mucous membrane contact. Substances may be deliberately introduced into the
body via contact with those mucous membranes that are externally accessible, such as
the eyes, ears, mouth, nose, and rectum. In drug abuse, amphetamines, cocaine, and
heroin are acquired by nasal insufflation.

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 ▪ Injection. Introduction of a compound directly into the body, generally by means
of a hypodermic syringe. The substance may be directed into a vein (intravenously) or
into a muscle (intramuscularly). It may also be administered under the skin
(subcutaneously) or into the skin (intradermal).
The bioavailability of a specific substance is the proportion of the original dose
that is absorbed and the rate at which this absorption takes place. This is influenced, for
example, by the chemical and physical attributes of the substance concerned and the
route of uptake used. Notably, substances absorbed from the intestine have to encounter
the liver (the key metabolic organ) before entering the general circulation.

The analysis of drugs and other poisons

1. The information sought by analysis: The investigation of a sample for drugs of
abuse and/or other poisonous substances targets to provide information about one or
more of the following: (i) whether a particular sample contains a controlled substance or
other poison; and (ii) whether the concentration of a particular substance within given
samples of body fluids and/or tissue is consistent with therapeutic administration (in the
case of drugs that have medicinal applications), intoxication or death by poisoning with
the substance concerned. For example, it can establish whether the concentration of
ethanol in a given sample of breath, blood, or urine exceeds the legally permissible limit
for driving.
▪ Qualitative analysis is the chemical analysis concerned with establishing the
identity of the analyte, the chemical species that is being analyzed for in the sample under
test.
▪ Quantitative analysis is the chemical analysis concerned with establishing the
concentration and/ or amount of the analyte and may help the courts to establish whether
the amount of heroin seized was consistent with personal use by the individual concerned
or with the intent to supply the drug to others.
2. The types of a sample that are analyzed:
▪ Bulk samples are those large enough to be weighed such as legal drugs in the form
of tablets, capsules, and samples of poisons or suspicious materials.
▪ Trace samples the amount is so small that it cannot be weighed or may well be
possible to establish its weight by means of quantitative chemical analysis. Both of these
are subjected to forensic analysis for drugs and other poisons to establish the identity,
composition, and/or quantity of one or more of their constituents.
▪ Distinguished among these are biological samples taken from people; the first of
these include those samples that can be acquired from living persons, in a particular
breath, blood, urine, stomach contents, hair, nail clippings, saliva and sweat, and those
tissues and body fluids that can be obtained during post-mortem examinations.
▪ The biological samples selected for analysis will differ from case to case and will
reveal: (i) those that are available such as a collection of samples in a living body; (ii)
those in which the analyte can readily be identified and/or quantified; (iii) those for
which there is widespread literature to help the explanation of the analytical results; (iv)
those that are acceptable for legal and ethical reasons, for instance, breath, blood, and
urine are all acceptable from a legal perspective for the determination of alcohol levels in
cases of possible drink-driving; (v) whether information about chronic or acute exposure
to the analyte is being sought.

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Methods of analysis
The methods used to analyze for drugs and other poisons all exploit the chemical,
physical, and/or biochemical properties of the analyte of interest that allow it to be
identified and/or quantified. Matrix is all of the samples except the chemical species being
analyzed.
▪ Readily made observations: These include the color and morphology of the sample
that may be seen with the naked eye and/or with the aid of a microscope. These
characteristics can be very informative. For example, shape, dimensions, color, and
manufacturers’ marks can be used to establish the identity of commercially produced
tablets and capsules. Yet, even in cases in which these observations lead to apparent
identification, care needs to be exercised. It is notable that even in cases in which
appearance is not adequate to categorize the material, valuable information can be
obtained from a visual examination.
▪ Presumptive tests: Bulk samples will, in many cases, be subjected to presumptive
tests or color tests, which are designed to rapidly and inexpensively specify the presence
of certain analytes but these tests are not completely specific.
o A small amount of the sample is treated with reagents that are known to produce
characteristic colors on reacting with the analyte of interest, thereby indicating a positive
result. These tests are seldom completely specific. That is, most will produce a positive
result with any one of a range of
different chemical species.
o The exact color produced will
vary from one chemical species to
another and the analysis of the color of
the reaction remains somewhat.
o The presence of impurities in
the sample may mask the color
produced or even produce their own
color reactions.
o The blank test consists of the
reagents only (with no sample
present) and helps to confirm that
contamination of the equipment used
has not occurred.
o Positive control contains both
the reagents and pure portion of the analyte that is being tested for and serves to prove
that the reagents do indeed lead to color production with the analyte of interest.
o Presumptive tests provide valued information that guides the analyst in the
choice of further tests that will confirm or refute the indications provided by the
presumptive tests. Lists of some presumptive tests commonly employed in the analysis
of drugs are shown in the table.
▪ Thin-layer chromatography. Both bulk samples and samples that contain trace
levels of drugs or other poisons may be analyzed by thin-layer chromatography (TLC).
This is also used to locate or visualize the analyte(s).
o Although TLC can improve the indications provided by presumptive tests,
confirmatory tests such as gas chromatography-mass spectrometry (GC–MS) will usually
have to be performed before the identity of the analyte can be clearly determined.

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 ▪ Immunoassay techniques are of significant value in the analysis of both trace and
bulk samples for drugs and other poisons. This is capable of quantifying this poison even
when chemically related herbicides are also present in the sample. And intended to detect
the presence of a range of chemically related compounds.

Elimination of Unabsorbed Poison. Elimination of the unabsorbed poisons depends

upon the mode of intake of the poison:
1. Inhaled poisons: The victim should immediately be shifted to fresh air. A clear
airway should be ensured and simulated respiration initiated simultaneously.
2. Injected poisons: The use of the strap above the spot of injection may slow the
absorption. The tourniquet or strap should be loosened every 10 minutes for 2 minutes,
imbibe the poison by creating incisions at the spot and use chemical antidotes, cold
packing, and vasoconstrictors.
3. Contact poisons: Affecting the skin, mucous membrane, any wound, or introduced
in the uterus or vagina they should be washed with water and neutralized by application
of the suitable chemical. If the poison is applied to eyes, skin, or wound, the poison is
removed with specific antidotes.
4. Ingested poisons: The poisons are most commonly ingested orally. Depending on
the time elapsed after ingestion. Much of the poison lying unabsorbed in the stomach
should be removed.
a. Stomach wash or gastric lavage: Gastric aspiration and lavage is the only suitable
method of emptying the stomach of an unconscious patient.
b. Emesis: Emetics are the substances, which produce a gagging effect. Most poisons
are themselves emetics and may cause vomiting.
c. Catharsis: Cathartics lessen the travel time of the poisonous substance in the
gastrointestinal tract but their efficacy in reducing the mortality or morbidity is not
established. There are two types of catharsis saline cathartics and saccharide cathartics.

Elimination of Absorbed Poison

The various methods of eliminating absorbed poisons from the body include the
following:
1. Hemodialysis: The three elementary components of hemodialysis are the blood
delivery system, the dialyzer itself, and the composition and method of delivery of the
dialysate.
2. Hemoperfusion: It is capable of removing many of the toxins that are not removed
well by hemodialysis.
3. Peritoneal dialysis works on the same principle as hemodialysis, allowing the
diffusion of toxins from mesenteric capillaries across the peritoneal membrane into the
dialysate dwelling in the peritoneal cavity.
4. Hemofiltration is performed similarly to hemodialysis except that the blood is
pumped through a hemofilter.
5. Plasmapheresis is a technique of separating cellular blood components from
plasma. The cells are re-suspended in either colloids or albumin, or fresh frozen plasma,
and then reinfused. It is very effective in eliminating toxic substances but exacts a heavy
toll.
Application of Antidotes

16
 Antidotes are remedies to neutralize the effects of poison. They are used because
the poison may not have been completely removed by emesis or lavage or the poison is
already absorbed or it has been administered by other routes than ingestion. According
to their mode of action, antidotes are divided into the following classes:
1. Mechanical or physical antidotes counteract the effect of poison mechanically
preventing their absorption without inactivating the damaging action of the poisons.
These are of the following types:
a. Activated charcoal confers a great adsorptive capacity to the toxic material.
b. Demulcents and Bulky foods. Demulcents produce a protective coating over the
mucous membrane of the stomach to protect it from the action of the poison. These are
milk, egg white, starch, milk of magnesia, and aluminum hydroxide gel. Bulky foods like
bananas, boiled rice, or potatoes.
2. Chemical antidotes. Disintegrate and inactivate poisons by undergoing chemical
reactions along with the poisons and forming harmless or insoluble compounds. In
corrosive alkali poisoning, weak vegetable acids like citric acid and acetic acid (vinegar,
lemon juice) may be used. Albumen is used for mercury poisoning as it precipitates
mercuric chloride. Tincture iodine or Lugol’s iodine in a solution of 15 drops in half a glass
of water precipitates lead, mercury, silver, alkaloids, and strychnine. Tannic acid (4%) or
strong tea or 1 teaspoon full of tannic acid dissolved in water is used to precipitate metals
like lead and mercury,
3. Physiological antidotes are basically of use when some of the poison has already
been absorbed into the circulation and antagonize the effects produced by the poisonous
substance. These are like atropine for organophosphorus compounds, physostigmine,
and neostigmine for Datura and barbiturate for strychnine.
4. Universal antidotes. It is used when the characteristics of poison are not apparent
or when a combination of poisons is suspected. It consists of a mixture of the following
substances: powdered animal charcoal (2 parts): adsorbs alkaloids; magnesium oxide (1
part): neutralizes acids; tannic acid or strong tea (1 part): alkaloids, glucosides, and
metals.
5. Chelating agents. These substances produce a firm non-ionized cyclic complex
(chelate) with cations. Such compounds can form stable, soluble non-toxic complexes
with calcium and certain heavy metals. They are BAL (dimercaprol), EDTA
(ethylenediaminetetracetic acid), penicillamine, and deferoxamine which are used in
heavy metal poisoning.

17
Assessing Learning

Activity 3

Name: ______________________________________________________ Rating: ____________

Course/Year/Section: ____________________________________ Date: ______________

Directions: Read each question carefully and write your final answer on the space
provided.
True or False. Write A if the statement is TRUE and B if it is FALSE.
___1. Toxins are poisons manufactured in laboratories.
___2. The major ways people are exposed to poisons are by ingesting them, inhaling them,
injecting them, or absorbing them through the skin or mucous membrane
___3. Neurotic poisons can be categorized as stimulants, depressants, and hallucinogens.
___4. Arsenic can cause symptoms of chronic poisoning.
___5. All substances are poisons and a right dose differentiates a poison from a remedy.
___6. A presumptive test like color tests can clearly identify the drug or poison present in
any sample.
___7. Chelating agents are specific antidotes for toxins such as alkaloids, aflatoxins, and
venom.
___8. All alcohols are toxic to the body.
___9. Mercury can cause symptoms of acute poisoning.
___10. Corrosive poisons are less active irritants and do not produce inflammation and
ulceration of the tissues.
ESSAY. Concisely discuss your answer.
1. Discuss the uptake of potentially toxic substances into the human body, their
distribution, and subsequent elimination. Within this context, explain what is meant by
the terms ‘absorption’ and ‘bioavailability’.
_____________________________________________________________________________________________________
_____________________________________________________________________________________________________
_____________________________________________________________________________________________________
_____________________________________________________________________________________________________
________________________________________________________
2. “The toxicity of a substance is determined not only by its inherent toxic properties
but also by a number of factors relating to the individual exposed to it.” Discuss. (Include
in your answer an explanation of the following conditions: sensitization, tolerance, and
idiosyncratic response.)
_____________________________________________________________________________________________________
_____________________________________________________________________________________________________
_____________________________________________________________________________________________________
_____________________________________________________________________________________________________
_____________________________________________________________________________________________________
_______________________________________________

18
UNIT IV. CLASSIFICATION OF DRUGS
INTRODUCTION
Nowadays, the term drug has been associated with a negative connotation as a dangerous
substance causing addiction and some other detrimental effects. Since the emergence of
“Oplan Tokhang”, the word “drugs” has been used on streets, social media, news, etc. with
utter disregard of its operational definition. In general, drugs are substances that bring
physical, physiological, psychological, and behavioral changes to a person. (Dascil-Cañete,
2014)

Drugs and Drug Abuse

As defined above, drugs are substances that can cause changes to a person’s health.
However, this definition does not reflect the difference between medicinal and dangerous
drugs.

Medicinal drugs are substances that are taken to relieve pain, and cure diseases. This is
usually prescribed by doctors though some can be bought without a prescription.
Dangerous drugs, on the other hand, are substances that specifically attack the central
nervous system causing damage at prolonged use. This is usually addictive and illegal.
(Dascil-Cañete, 2014)

All medicines are drugs but not all drugs are medicine.

Drug abuse is the prolonged exposure of a person to drugs or excessive use of drugs which
are beyond its intended use. Abuse does not only include illegal drugs but may also apply
to drugs (medicine) that are legally available on the market. (Drugs of Abuse: A DEA
resource guide, 2017)

CLASSIFICATION OF DRUGS
Drugs can be classified according to:
1. Origin
2. Legal Classification
3. Effects (Pharmacological Classification)
Drugs According to origin:

A. Natural Drugs are active components extracted from a natural source such as
plants, and other living systems. Examples of this are marijuana and coca leaves.
B. Synthetic drugs are artificially made substances. These are produced in
laboratories from mixing different chemical compounds. Example of this
Methamphetamine and barbiturates (Rory Keane, 2006)
Drugs according to Legal Classification.
Drugs may be classified according to local law

19
 A. Presidential Decree 1619 of 1979
- Prohibiting the possession of unauthorized sales to minor of the volatile
substances for the purpose of inducing intoxication. (Official Gazzette, 2020)

B. Comprehensive Dangerous Drug Act of 2002 (RA 9165).

-In RA 9165, the term prohibited and regulated was changed into dangerous
drugs with three classifications.

a. Immediate precursors are substances incorporated in the production of final

molecules of the product.
b. Essential Chemicals are substances which are used as reagents or solvents in
the production process of an illegal drug. (Dascil-Cañete, 2014)
c. Narcotic, psychotropic, and designer drugs.

-The term “narcotics” pertains to stimulant drugs or substances coming

from opiate, cannabis (marijuana), and cocaine. While the “psychotropic drugs”
denotes “mind-altering drugs. Examples of these are Lysergic Acid Diethylamide
(LSD), amphetamines, barbiturates, and other designer drugs. (A Forensic Guide
for Crime Investigators, 2017)

C. Dangerous Drug Act of 1972 (RA 6425)

- In RA 6425, dangerous drugs are classified into three categories:

a. Prohibited Drug which includes natural or synthetic drugs with the

physiological effects of narcotics or hallucinogenic drugs. Example of these are opium
and its active ingredients and derivatives, coca leaf and its derivatives, Indian hemp,
and its derivatives
b. Regulated Drug includes self-inducing sedatives, hypnotic drugs, and other
drugs with similar physiological effects to amphetamine. Example of these are
secobarbital, benzedrine, and nitrazepam
c. Volatile Substances these are substances mostly liquid which are capable of
releasing toxic vapors or fumes to induce intoxication, excitement, etc. (Dascil-
Cañete, 2014)

Drugs according to its Effects (Pharmacological Effects)

The drugs may be classified according to their effects on a person’s body such as
stimulants, hallucinogens, depressants, and inhalants.

a. Stimulants are drugs that increase alertness and speed up the body’s system.
These substances produce a sense of excitement, enhance self-esteem, improve
physical and mental activity, and create a sense of “high”. Most stimulants are sold

20
 as pills, tablets powder, and injectable (Drugs of Abuse: A DEA resource guide,
2017).
Examples of this are amphetamines, methamphetamine, cocaine, and khat.
Amphetamine

One of the most commonly sold

stimulants is an amphetamine, but some are
legally prescribed to treat ADHD. It was sold in
the 1930s as Benzidrine an over the counter
inhaler to treat nasal congestion but due to
abuse in use and uncontrolled production, it
was banned and was then declared regulated.
(Drugs of Abuse: A DEA resource guide, 2017)
This drug can be easily made from an
over the counter drug pseudoephedrine which
is a common ingredient of cold medicine.
The common street names for amphetamine are bennies, crank, ice, and
uppers. (Dascil-Cañete, 2014).
To know more about Amphetamine please follow this link:
https://www.youtube.com/watch?v=cl8Nhwn4Dz8

Cocaine
Cocaine is another stimulant drug commonly sold on the
streets. It creates an intense euphoria feeling with a strong
tendency to be addictive. The dose and mode of delivery
determine how fast the drug reaches the brain creating a
euphoria feeling known as “rush”. Cocaine is a natural stimulant
derived from coca leaves which commonly grow in areas like
Columbia and Peru. The plant extract undergoes series of
chemical transformation to achieve a crystal-like appearance.
(Drugs of Abuse: A DEA resource guide,
2017).

The common street name for cocaine is crack,

coke flakes and snow.

To know more about cocaine please follow

this link: https://www.youtube.com/watch?v=7tlwW2DKzlc

21
Methamphetamine

Methamphetamine (meth) is a synthetic stimulant

drug. It is a potent central nervous system stimulant
that can be taken by oral ingestion or by snorting. It
is said that it releases a very high level of dopamine,
a neurotransmitter, upon reaching the brain creating
the feeling of pleasure. Other modes of intake are
though injection and smoking. (Drugs of Abuse: A
DEA resource guide, 2017)
The major effects of meth in our
body are an increase in physical
activity and heart rate thus
creating hyperthermia or
overheating. This is the reason
why a person taking meth tends
to feel hotter even on cold nights.
It is also reported that it increases
wakefulness and decreases appetite. (Drugs of Abuse: A DEA resource guide, 2017)

Some common street names for methamphetamine are meth, crystal, meth, shabu, shabs,
sha, and ice. (Dascil-Cañete, 2014)

To know more about methamphetamine please follow this link:

https://www.youtube.com/watch?v=8bvcfc7xVnU

b. Hallucinogens
Hallucinogens are drugs that affect sensation, thinking, self-awareness, and
emotion. (Dascil-Cañete, 2014). These drugs can be derived naturally from plants
and some fungi or can be made synthetically in clandestine laboratories. These
drugs can tale a variety of forms from tablets, pills, and liquid injectable. (Drugs
of Abuse: A DEA resource guide, 2017)
Examples of these are ecstasy, Marijuana, and LSD.

Ecstacy

Ecstasy is both a psychedelic and stimulant

drug that produces an energizing effect with
distortion of time and perception. (Drugs of
Abuse: A DEA resource guide, 2017)

22
 Its chemical name is 3,4
methylenedioxymethamphetamine and
is synthetically made in laboratories.
Manufacturers may add anything
various drugs like amphetamine and
caffeine (Drugs of Abuse: A DEA
resource guide, 2017)

It is commonly found in pill form with attractive color and designs to target party
goers like young adults and adolescents. Some common names for this drug are
XTC, adam, essence, E and X. (Dascil-Cañete, 2014)

Marijuana
Marijuana is a psychoactive drug
derived from the Cannabis native
plant. It is commonly grown in South
America, Canada, Mexico, and Asia.
Some of the effects of marijuana are
induced pleasure, increased
memory, and concentration though
some of the reported side effects are
memory loss, difficulty in
concentration, and distorted
perception. (Drugs of Abuse: A DEA resource guide, 2017)

Marijuana is commonly smoked as a crushed brown dried leaf inserted into pipes
or cigarettes. It also reported that it is included in foods and some are made into
candies in some areas where it is legal to use. Commons names for marijuana are
mary jane, flower, damo, brownies, joint, and dope. (Dascil-Cañete, 2014)
To know more about marijuana please follow this link:
https://www.youtube.com/watch?v=r2_skiJAHsQ

c. Depressants

Depressants are drugs that depress or lower the functions of the central nervous
system. (Dascil-Cañete, 2014). These drugs induce sleep ad relieves muscle
spasm and anxiety. Barbiturates are older known depressant though it is known
to be unsafe and has a tendency to cause coma and death. It was later replaced by
benzodiazepines which were less potent than barbiturates and available upon
prescription. Examples of this are Valium, Klonopin, Ativan, and Xanax. It is
available in pill, syrups, and injectable liquids. (Drugs of Abuse: A DEA resource
guide, 2017)

23
 Depressants can be classified according to types; narcotics, tranquilizers, and
sedatives.
Narcotics are drugs that induce sleep and relieve pain examples of these are
Opium, Opiates Heroin, and Morphine. Tranquilizer is a type of depressant that
relaxes the muscle and reduces anxiety. Sedatives and hypnotics are drugs that
aids sleep and have calming effects. It is commonly prescribed to treat sleep
disorders and in high doses acts as a anesthetics. Examples of these are
barbiturates and alcohol. (Dascil-Cañete, 2014)
To know more about depressant please follow this link:
https://www.youtube.com/watch?v=dOlkogaWF3M
d. Inhalants
Inhalants are liquid, solid, or mixed substances that commonly found on
households that are capable of releasing vapors or fumes that when inhaled
creates a psychoactive or mind-altering effect.. Examples of this are solvent, glue,
gasoline paint, and cleaning liquids. This is known to cause brain damage affecting
thinking and perception. (Dascil-Cañete, 2014).

To know more about inhalants please follow this link:

https://www.youtube.com/watch?v=smqpu_TErl

References
A Forensic Guide for Crime Investigators. (2017, January 4). Retrieved from
National Institute of Criminology and Forensic Science:
http://nicfs.gov.in/?p=15475
Dascil-Cañete, A. M. (2014). Forensic Chemistry and Toxicology. Quezon City:
Wiseman's Books Trading, Inc.
Drugs of Abuse: A DEA resource guide. (2017). Drug Enforcement Agency: US
Dept. of Justice.
Official Gazzette. (2020, July 20). Retrieved from Ofiicial Gazzete Open Data
Potal: https://www.officialgazette.gov.ph/1979/07/23/presidential-
decree-no-1619-s-1979/
Rory Keane, S. R. (2006). Understanding substances and Substance Use. Dublin
West Education Center.

24
Assessing Learning

Activity 4

Name: ______________________________________________________ Rating: ____________

Course/Year/Section: ____________________________________ Date: ______________

Please complete the following table

Substance Example What do they What are the

do? side effects?

Stimulants

Hallucinogens

Depressant

Inhalants

25
UNIT V. DNA
At the end of this unit, I will be able to:
1. discuss the chemistry of the genetic information carrier.

Central Dogma of Molecular Biology

• Describes the interrelationship of 3 major biomolecules: DNA, RNA, proteins (How

genes control which and when proteins may be synthesized by the body).
• Comprised of the 3 major processes in gene expression:
1. Replication (DNA --> DNA)
▪ The copying of parental DNA to form daughter DNA molecules with a
completely identical sequence
2. Transcription (DNA --> mRNA)
▪ The process by which parts of the genetic message encoded in DNA are copied
precisely into RNA
3. Translation (mRNA --> protein)
▪ The genetic message encoded in a messenger RNA (mRNA) is translated by
ribosomes into a polypeptide with a particular sequence of amino acids.

Figure 5.1 The Central Dogma of Molecular Biology.

• Chromosomal DNA: located in the chromosomes, which are within the nucleus;
dictates activities of the cell.
• Mitochondrial DNA: located in the mitochondrion; encodes mitochondrial proteins.

Figure 5.2 The Human Genome. Genes are found within DNA, which is condensed into
chromosomes, which are mostly kept in the nucleus of the cell.

• GENES: segments of DNA that code for polypeptide chains and RNAs

26
 o Components of the information unit of living organisms.
o The entire DNA that encodes the primary sequence of some final gene product,
which can be either a polypeptide or RNA with a structural or catalytic function.
o Specific unit in the chromosome that will encode for a specific protein; one
chromosome will contain more than 1 gene
o The number of genes depends on how many chromosomes an organism has

Table 1. Comparison of Chromosomal Composition

Organism Total Base # of # of genes

Pairs chromosomes
E. coli 4,639,221 1 4,405
Yeast 12,068,000 16 6,200
Fruit Fly 180,000,000 18 13,600
Mouse 2,500,000,000 40 30K-35K
Human 3,200,000,000 46 30K-35K

▪ Simplest is the E coli genome containing 4.6 million bases that will end up being
stored only in one chromosome
▪ A diploid cell of a human individual will contain as much as 6.4 billion base
pairs that are going to be distributed and compacted into 46 chromosomes:
o 22 pairs will be the autosomes/somatic chromosomes
o 1 pair remain as the XY chromosome (sex-dependent)
o Note that each chromosome will not contain 6.4 billion bps. These are
distributed over the 23 pairs unevenly (some will have more, some will
have less).

Figure 5.3 Molecular Nature of a Gene.

Nucleotides

• Monomers of nucleic acids (DNA and RNA)

• Composed of:
o Phosphate group: bonded to C5’ (PHOSPHOESTER BOND)
o Nitrogenous base: bonded to C1’ (N-GLYCOSIDIC BOND)
▪ Purines (via N3) – Adenine, Guanine

27
 ▪ Pyrimidines (via N9) – Cytosine, Uracil, Thymine
o Sugar
▪ RIBOSE – sugar in RNA; has –OH at 2’ position
▪ DEOXYRIBOSE – sugar in DNA; does not have –OH at 2’
• The polymerization of nucleotides give rise to nucleic acids
o The linkage between nucleotides involves the formation of 2 ester bonds by
phosphoric acid. The OH groups to which the phosphoric acid is esterified are
those bonded to the 3’ and 5’ carbons of the sugar on adjacent residues.
o The resulting repeated linkage is a 3’, 5’-PHOSPHODIESTER BOND.
▪ The chain of sugars linked by PHOSPHODIESTER BRIDGES is referred to as the
BACKBONE of the nucleic acid (Figure 5).
• Nucleotide residues are numbered from the 5’ end, which normally carries a
phosphate group, to the 3’ end, which normally has a free OH group.
DNA:

The chemical basis of heredity and is organized into genes, the fundamental units of
genetic information contain:
▪ CODING sequences, which would encode the primary structure (sequence) of
proteins and RNAs
▪ REGULATORY sequences which would determine what will happen in the
central dogma; provide signals that may:
a.) denote the beginning of the end of genes, or
b.) influence the transcription of genes or function as initiation points for
replication or recombination
• Human DNA is made up of at least 6 million base pairs
• The double-stranded polymer of nucleotides with the two strands running anti-
parallel (in opposite directions) to each other
• Base pairing (ONE PURINE to ONE PYRIMIDINE)
o Follows a specific rule (Watson-Crick base pairing) to maintain the regular helical
structure independent of the base sequence
▪ Adenine (A) – Thymine (T) (*replaced with U in RNA)
▪ Guanine (G) – Cytosine (C)
• Chargaff’s rules:
a) The base composition of DNA varies from one species to another.
b) DNA specimens from the same species will contain the same base composition
despite coming from different tissues.
c) Base composition doesn’t change with age, nutritional
state, or environment.
d) # of adenine (A) = # of thymine (T),
# of guanine (G) = # of cytosine (G)
# of (A+G) = # of (T+C)
RNA

• Single-stranded molecule
• RNA vs DNA (differences in covalent structure)
o BOTH have the same 3’ 5’ phosphodiester backbone

28
 o The difference lies in sugar units and bases in sequence:
a) Ribose in RNA; Deoxyribose in DNA
▪ Ribose contains a 2’-hydroxyl group
▪ In addition to the standard 3’ 5’ linkage, a 2’ 5’ linkage is possible for RNA.
This later linkage is important in the removal of introns and the joining of exons
for the formation of mature RNA.
b) Uracil (U) in RNA; Thymine (T) in DNA
• 3 types of RNA:
1) mRNA (messenger RNA) – template for protein synthesis or translation
2) tRNA (transfer RNA) – carries amino acids in an activated
form to the ribosome for the peptide-bond formation
3) rRNA (ribosomal RNA) – where polypeptide chain is
synthesize
Table 2. Comparison of DNA vs RNA

DNA RNA
Sugar Deoxyribose Ribose
Major Purines Adenine and Guanine Adenine and Guanine

Major Pyrimidines Thymine and Cytosine Cytosine and Uracil

Comprise about 1% of the About 5-10%

total weight of the cell
Found primarily in the Exists in 3 major forms
chromosomal form in the (rRNA, tRNA, mRNA)
cell’s nucleus

References
Devlin, Thomas M. Textbook of Biochemistry with Clinical Correlations. New York: John
Wiley & Sons Inc. 7th ed.
Nelson, David L., Michael M. Cox, and Albert L. Lehninger. Lehninger: Principles of
Biochemistry. New York: W. H. Freeman.

29
Assessing Learning

Activity 5

Name: ______________________________________________________ Rating: ____________

Course/Year/Section: ____________________________________ Date: ______________

I. Match the following definitions to the correct vocabulary term.

____ 1.) DNA A) carries genetic information.

____ 2.) Gene B.) controls protein production.
____ 3.) Chromosome C.) helical molecule
II. Write the letter on the line of the choice that best answers each question.
_____ 4.) How many nitrogenous bases are there in DNA?
A.) 1 D.) 4
B.) 2
C.) 3
_____ 5.) Which of the following is not a nitrogenous base found in DNA?
A.) Adenine B.) Cytosine
C.) Guanine D.) Uracil
_____ 6.) DNA is made up of Nitrogenous base pairs. The bases bond in which of the following
patterns?
A.) Adenine – Thymine; Cytosine – Guanine
B.) Adenine – Cytosine; Guanine – Thymine
C.) Adenine – Guanine; Cytosine – Thymine
D.) Adenine – Guanine; Cytosine – Taurine

_____ 7.) DNA looks like a “twisted ladder”. If the “rungs” of the ladder are made up of nitrogenous
bases, what are the handrails or backbone of the ladder made up of?

A.) Sugars and Salts B.) Phosphates

C.) Sugars D.) Sugars and Phosphates

_____ 8.) Where is DNA located in the cell?

A.) Mitochondria B.) Cell Membrane

C.) Nucleus D.) Golgi Body
_____ 9.) How many chromosomes do we get from our mother?

A.) 46 B.) 23
C.) 22 D.) 20

_____ 10.) Who is credited with discovering the structure of DNA?

A.) Crick and Neck B.) Watson and Crick

C.) Watson and Franklin D.) Holmes and Watson

30
UNIT VI: BLOOD AND BLOOD SPATTER
Learning Objectives:
After reading this unit, I will be able to:
1. explain the composition of blood;
2. describe the functions of blood cells;
3. describe a brief history of the use of blood and blood spatter analysis in forensics;
4. describe how to determine the blood type of a sample of blood;
5. describe how to screen for the presence of human blood.
6. conduct a blood-spatter analysis; and
7. use blood-spatter evidence to recreate the events at a crime scene.

Composition of Blood
Blood is a circulating tissue containing three types of
cells: red blood cells, white blood cells, and platelets. These
cells are suspended in a liquid known as plasma. Plasma is like
saltwater in composition. It brings dissolved proteins, such as
antibodies, hormones, and clotting factors, and nutrients such
as glucose, amino acids, salts, and minerals.
Blood Cells. Each blood cell performs a different function. (i)
Red blood cells (RBC or erythrocytes) transmit respiratory gases, mainly oxygen and carbon
dioxide. The hemoglobin in red blood cells is an iron-containing protein that binds to oxygen
in the lungs and transports the oxygen to cells in all
the tissues in the body. Hemoglobin in red blood
cells is also in charge of the red color in blood. (ii)
White blood cells (WBC or leukocytes) fight disease
and foreign invaders. (iii) Platelets (thrombocytes)
aid in blood clotting and are involved in repairing
damaged blood vessels. Our bodies have the ability
to discriminate between their own cells and
molecules (self) and foreign invaders (non-self).
The immune system’s job is to protect our bodies by
recognizing cells or molecules that are foreign, such
as viruses, bacteria, and other parasites. When the
immune system identifies the existence of attacking foreign molecules, white blood cells,
which travel all over the body, concentrate in the location of the invading material—whether
it is a virus, bacteria, or protein. The white blood cells engulf and digest the invader. Other
white blood cells secrete proteins, known as antibodies, which assist in the immune response.
Because white blood cells are the only type of blood cell that contains a nucleus, they are the
only blood cells that can be used as a source of DNA for DNA profiling. Serology involves the
study of body fluids and stains at the crime scene, particularly blood.

Blood Typing
Blood typing is less expensive and faster for studying blood evidence than is DNA
profiling. Because many different people share the same type, this blood evidence is
considered to be class evidence. By typing the blood found at a crime scene, it is likely to
connect a suspect to a crime scene or to eliminate a suspect. Though, matching blood types
does not verify guiltiness because many people share the same blood type.

31
 ▪ Discovery of Blood Types. In 1900, Karl Landsteiner establish that blood from one
person did not always freely mix with blood from another person. Instead, clumping might
occur, which could result in death. The occurrence or lack of specific proteins found
implanted within the cell or plasma membranes of red blood cells determine a person’s blood
type. Landsteiner in 1901, described the A and B proteins found on the surface of red blood
cells. Rh factor which is another type of RBC proteins were also identified. The presence or
absence of these cell-surface proteins gives rise to our present system of blood typing. An
antibody reaction test is used to identify each blood type.
▪ ABO Blood System. ABO blood type
is a genetic example of multiple alleles. A and
B proteins are found on the surface of some
RBC. If a person’s blood has only protein A,
then he or she has blood type A. If the blood
has only protein B, then the person has blood
type B. If the person’s blood has both the A
and the B proteins, then he or she has blood
type AB. The blood of some people lacks both the A and the B proteins. This blood type is
designated as type O blood.
▪ Rh Factor. Alexander Weiner in 1940,
noticed another type of RBC protein while he was
working with Rhesus monkeys. Eighty-five percent of
the human population has a protein called Rh factor
on their RBC, it can be designated as Rh+ (positive),
while blood that does not have this factor is
designated Rh– (negative).
▪ The naming of Blood Types. The presence
or absence of the AB and Rh proteins determine a person’s blood type. The earliest
accomplished blood transfusions occasionally helped the recipient of the blood, but many
times the transfusion killed the person. In the early days of transfusion, doctors were not
conscious that people have diverse blood types. The presence of different blood types was
not discovered until 1901. When a person receives a blood protein that is foreign to him or
her, antibodies will cause the blood to clump and may cause death.

Antibodies
To assist WBC, recognize foreign
proteins, B-lymphocytes, a specialized
type of WBC, discharge antibodies. The
antibodies are Y-shaped protein
molecules that bind to the molecular
shape of an antigen, fitting like two
complementary puzzle pieces. The
binding site of the antibody is located on
the tip of the Y-shaped molecule. The
antibody recognizes a foreign substance
as an invader and attaches to it.
▪ Antigen-Antibody Response. A process when a foreign invader is recognized by the
immune system; an attack is launched against that invader. WBC recognizes a substance as
foreign and try to destroy it. The foreign invaders may be viruses, bacteria, or even the red
blood cell proteins from a person with a different blood type. For example, if a person with
type A blood receives a transfusion containing type B blood, foreign type B antigens alert the

32
immune system to the presence of an invader. The body responds in two ways: (1) B
lymphocyte makes specific antibodies against that protein; these antibodies are Y-shaped and
have specific binding sites that attach to foreign proteins; and (2) phagocytes, a type of white
blood cells, engulf the invader.
▪ Agglutination. There are more than
300 known blood group proteins and more
than 1 million different protein-binding sites
on each RBC and many of these proteins are
on the surface of the cell membrane. When
one arm of the Y-shaped antibody attaches to
the RBC, the second arm of the Y attaches to
another RBC, which results in agglutination
or clumping, of the RBC. If this clumping
occurs within the circulatory system, a recipient person during a blood transfusion, blood
could stop to flow. The blood vessels will become clogged by clumped red blood cells. Without
blood circulation, the cells of the body cannot receive oxygen or eliminate carbon dioxide,
and the person will die.
▪ Blood Typing Tests. Blood typing
is a way to classify and match blood
samples. This is vital before a blood
transfusion to ensure that the transfused
blood does not contain any foreign blood
proteins and a person’s blood is verified
for the presence of three RBC proteins: A,
B, and Rh. Three separate tests are
performed. The patient’s blood is mixed
with antibodies that bind to the A protein. If the patient’s blood clumps or agglutinates, that
means that the person’s blood contains protein A. If the blood being tested does not respond
with the antibodies that bind the A protein, then that person’s blood does not contain protein
A. Similar testing is done with antibodies to protein B and the Rh factor. If the blood clumps
or agglutinates in the presence of these antibodies, it means that those proteins are present
in the blood. In the absence of these blood proteins, no clumping will occur.

People with blood type O are called universal donors because they can give blood to
any blood type. People with blood type AB are called universal recipients because they can
receive any blood type. Rh+ can receive Rh+ or Rh- while Rh- can only receive Rh-. Genotypes
can be homozygous, in which both copies of a given gene have the same allele (I AIA) or
heterozygous, which has two different alleles of a given gene (IAi). For the blood type A (IAi)
genotype, “IA” is the dominant allele while “i” is the recessive allele, the same applies to blood
type B.

Additional Blood Proteins and Probability

Many other blood proteins have been identified. The presence or absence of these
proteins is inherited from our parents. In addition to A, B, and Rh protein, people can have M

33
proteins and N proteins. Approximately 30 percent of the population is pure MM, inheriting
a gene for the M protein from each parent. Twenty-two percent of the population is pure NN,
inheriting a gene for the N protein from each parent. The remaining 48 percent of the
population have MN, inheriting a gene for M from one parent and a gene for N from the other
parent.
Additional enzymes and proteins have been found in the blood, which is significant
for identification purposes. They include phosphoglucomutase (PGM), adenylate kinase (AK),
adenosine deaminase (ADA), esterase D (EsD), glucose-6-phosphate dehydrogenase (G-6-
PD), polymorphic proteins: Group-specific Components (Gc), and haptoglobins (Hp).

Blood Spatter
When a wound is inflicted and blood leaves the body, a blood-spatter pattern may be
created and a single stain or drop of blood does not considered spatter. Instead, a grouping
of bloodstains composes a blood-spatter pattern. This pattern can help rebuild the series of
events surrounding a shooting, stabbing, or beating.
▪ History of blood-spatter analysis. Pitoroski (1894) composed the earliest reference
to blood-spatter analysis. Balthazard (1939) was the first researcher to analyze the meaning
of the spatter pattern. In 1955, blood-spatter evidence was used by the defense in the Sam
Shepard case, helping to exonerate him. In 1971, Dr. Herbert MacDonnell used blood-spatter
analysis as a tool in modern forensic examinations. Today, blood-spatter evidence is used to
explain events at a violent scene.
▪ Blood-Spatter Analysis. Given the blood-spatter patterns, it is likely to determine
the direction the blood was traveling, the angle of impact, and the point of origin of the blood.
Blood-spatter patterns can help determine the manner of death, based on the blood velocity.
Instructions on blood-spatter analysis are provided within
each activity.
o Did you ever wonder why blood forms droplets as
it falls from a wound? If blood is a mixture, then why
doesn’t it separate in the air before it hits the ground or an
object? Why does a drop of blood have a curved surface
when it lands on a flat surface instead of spreading out
flat? The answers to these questions have to do with what
happens when the forces of gravity, cohesion, adhesion,
and surface tension act on blood.
o Blood is a thick mixture of blood cells and plasma.
When a person is wounded and has a hemorrhage, gravity
acts on the blood, (i) pulling it down toward the ground.
The blood droplet has a tendency to (ii) become longer
than it is wide as a result of gravity. Blood is (ii) cohesive,
means that the blood mixture is attracted to similar blood
mixtures, and tends to stick together and not separate as it
falls.
o The effect of the downward force of the gravity combined with the cohesive force of
the blood results in a net effect on the blood droplet as it falls. Thus, the blood maintains a
circular or round appearance.

34
 o When a drop of blood falls on a flat surface, the blood
drop will have a curved surface. The (iv) blood drop does not
totally flatten out because of the cohesive nature of blood
causing the blood to pull together and resist flattening out on a
surface. The result is that the surface of the blood is elastic,
giving the top of the blood spatter a spherical appearance.
o If any of the blood does overcome cohesion and
separate from the main droplet of blood, it will (v) form small
secondary droplets known as satellites.
o If blood is dropped onto a smooth surface, such as
glass or marble, the edge of the blood drop looks smooth and
circular. But, if the blood lands on a porous surface, such as
wood or ceiling tile, then the edge of the drop of blood may
form small spikes or extensions. Notice that spikes are still
connected to the main droplet of blood, whereas satellites are
totally separated.
o Dr. John Glaister (1902) first described the six
patterns into which blood spatters could be classified. They
include:
1. Passive fall occurs when blood falling directly to the floor at a 90-degree angle will
produce circular drops, with secondary satellites being more produced if the surface hit is
textured.
2. Arterial spurts or gushes typically found on walls or ceilings are caused by the
pumping action of the heart.
3. Splashes are shaped like exclamation points. The shape and position of the spatter
pattern can help locate the position of the victim at the time of the attack.
4. Smears are left by a bleeding victim depositing blood as he or she touches or brushes
against a wall or furniture.
5. Trails of blood can be left by a bleeding victim as he or she moves from one location
to another. The droplets could be round or smeared or even appear as spurts.
6. Pools of blood form around a victim who is bleeding heavily and remains in one place.
If the bleeding victim moves to another location, there may appear to be droplets or smearing
connecting the first location with a second.
The size and shape of blood droplets aid categorize the path from which the blood
originated. For example, blood dripping downward at a 90-degree angle will produce round
droplets. Blood droplets with tails or satellite droplets aid us to define the direction from
which the blood originated.
Spatter patterns can help the investigator determine the type of wound. A fine-mist
spatter pattern is produced by a high-velocity impact, such as a gunshot wound. The lower-
velocity pattern can be observed by a beating with a pipe which produces blood cast off. Voids
or empty spaces in the spatter pattern could help determine the presence of a person or
object moved after the attack. By means of the spatter pattern to conclude the angle of impact
of several blood droplets, the examiner can determine the point of impact or convergence, a
two-dimensional representation of the location of the victim at the time of the injury.

Examination of Directionality of Blood

The shape of an individual drop of blood provides clues to the direction from where
the blood originated.

35
 ▪ A circular drop of blood (width and length are equal) indicates that the blood fell
straight down (90-degree angle of impact). This is typical if the blood was passively produced
(without any force). This would be typical of blood dripping from a wound.
▪ When a blood drop is elongated (longer
than it is wide), it is likely to define the direction
the blood was traveling when it struck a surface.
As moving blood strikes a surface, several forces
affect the droplet of blood. These forces are (i)
cohesion is a force between two similar
substances; (ii) adhesion is a force between two
unlike surfaces, such as blood and the surface of a
wall; and (iii) surface tension is an elastic characteristic along the outer edge of a liquid caused
by the attraction of like molecules.
▪ When blood comes into contact with another
surface, the blood tends to adhere or stick to it. As a
result, the point of impact may appear to be darker
and wider than the rest of the drop of blood spatter
(pattern of cast-off blood).
▪ Momentum tends to keep the blood moving
in the direction it was traveling. As it travels, some of
the blood follows to the new surface. However,
because of cohesion, most of the blood tends to stay as one drop. As blood droplets move
away from their source, the blood droplet elongates and may produce a thinner tail-like
appearance. The tail points in the direction of the blood’s movement. Smaller satellite or
secondary droplets may break away from the main drop of blood. These satellites will appear
in front of the moving droplet of blood. Note that satellites are not connected to the main drop
of blood.

Lines of Convergence
The place of the source of blood can be
determined if there are at least two drops of
blood spatter. By drawing straight lines down
the long axis of the blood spatter and noting
where the lines intersect, this will indicate the
lines of convergence. When there are much
blood spatters the area where the lines of
convergence meet are where the source of
blood originated. One can draw a small circle
around this intersecting area to note the area of convergence. The circle locates the area of
convergence and identifies in a two-dimensional view the location of the source of the blood.

Blood Spatter Types. Blood spatter may also be classified by its speed or velocity on
impacting a surface.

36
Crime-Scene Investigation of Blood
▪ In an effort to hide evidence, a culprit may try to eliminate blood sign by cleaning the
area. Even though a room may look flawlessly clean and totally free of blood after a full
washing of the walls and floor, blood evidence still remains. Red blood cells are anucleate,
they lack a cell nucleus so they contain much more cytoplasm than most other cells. This
cytoplasm is largely filled with a molecule called hemoglobin, which carries iron and bind to
oxygen molecules. Given the abundance of hemoglobin in the blood, and its unique
characteristics, it is often protein used to identify blood at the crime scene. To detect
hemoglobin, an investigator mixes Luminol powder with hydrogen peroxide in a spray bottle.
The mixture is then sprayed on the area to be examined for blood. The iron from the
hemoglobin, acting as a catalyst, speeds up the reaction between the peroxide and the
Luminol. As the reaction progresses, light is generated for about 30 seconds on the surface of
the blood sample.
▪ Luminol can also be used to detect blood in darkened rooms or at night. Even with
the most thorough cleaning, blood residue is difficult to remove. Traces of blood may adhere
to surfaces for years, even after cleaning. Luminol works best in an area where an attempt
has been made to clean up the blood.
▪ Once found, there are several steps used in processing a bloodstain, and each can
provide a different kind of critical information:
1. Confirm the stain is blood. At a crime scene, clothing is found with a red stain. Could
ketchup, ink, or any other red substance cause the red stain? Before trying to collect the blood,
it is first necessary to confirm if it is actually blood.
▪ Visual examination is valuable to determine whether the given stain is of blood or not.
There is no difficulty in fresh but old stains may not be identifiable. This determines the
number and size of stains; a rough estimate of the amount of bloodshed; the direction from
which the blood has fallen is found from the tip of the elongated stain; the shape of the stain
indicates the height from which the blood has fallen; from the positions of the stains, the
movement of the victim and the culprit can be determined.
▪ Ultraviolet or infrared rays reveal washed or invisible blood stains from clothes,
furniture, earth, doors, etc. They also reveal stains on colored garments or on painted
surfaces. They help to reveal minute blood traces.
▪ Spectrophotometry and chromatography are also used to identify blood.
▪ Color tests are the first series of tests employed after the visual study of stains. If a
stain gives positive color reactions in any of the two-color tests, the stain is possibly a
bloodstain. If it fails to give color reactions in all probability it is not a bloodstain or the same
cannot be established.

37
 o Benzidine reaction: Benzidine (0.1g) and dry sodium perborate (0.1g) are dissolved
in glacial acetic acid (10ml) and sprayed over the stain. A strong blue color indicates blood.
o Leucomalachite Green reaction: The reagent is prepared by dissolving
leucomalachite green (0.1g), sodium perborate (0.3g) in 65% glacial acetic acid. The reagent
is applied to the stain. Intense green color indicates blood.
o Phenolphthalein reaction: Phenolphthalein (1g) is reduced and dissolved in acetic
acid (100ml). Sodium perborate (1.4g) is dissolved in the solution and applied to the
bloodstain. Pink coloration indicates blood.
o Luminol test: Luminol is a chemical that has been used to locate the bloodstains. The
articles suspected to bear stains are sprayed with luminol. It reacts with the blood to give
fluorescence. The bloodstains are thus made visible. Even decomposed blood reacts with the
reagent. The reagent does not interfere with subsequent blood tests. It is prepared by
dissolving sodium perborate (0.7g) in water (100ml) and adding 3- arninophthalhydrazide
(0.1g) and sodium carbonate (5.0g) to the solution. The solution is sprayed upon the article
with glass sprayers in a dark room. Blood gives strong luminescence.
o Crystal test: Two crystal tests are commonly employed. They are specific for blood
but they are not sensitive. They often fail if the conditions are not rigidly controlled or if the
blood is disintegrated or contaminated. (i) Teichmann Test: Take a dry crust or smear of
blood on a slide. Put a drop of potassium iodide, bromide, or chloride (0.1gm) solution in
100ml glacial acetic acid over the blood and cover it with a coverslip. Warm the slide gently
till it gives out bubbles. Typically, hemin crystals are observed under a microscope. Heating
may have to be repeated a number of times. (ii) Takayama Test: It is also performed similarly.
The reagent is prepared from one volume of glucose solution (10%), one volume of potassium
hydroxide solution (10%), and two volumes of pyridine which are dissolved in six volumes
of water.
o Precipitin reaction: the most widely used test and the reaction is performed to
identify the species of the origin of the bloodstains. It is a very delicate test and requires only
small amounts of blood. A dilute blood solution is used for precipitin reaction. The antisera
are not diluted. The blood solution and antisera should be free from turbidity and
contamination.
o Kastle-Meyer test. Phenolphthalein and hydrogen peroxide are used. If blood is
present, a dark pink color is produced.
2. Confirm the blood is human.
▪ Microscopic examination of bloodstain is important in many situations such as in
determining species of origin of fresh bloodstains and the part of the body from which the
blood has come.
▪ Electrophoresis used for the separation of various proteins, adopted to study the
body proteins, and important to distinguish between blood samples.
▪ One test known as an ELISA test (Enzyme-Linked Immunosorbent Assay) involves an
antibody–protein reaction.
2. Determine the blood type. Blood collected from a crime scene is tested using specific
antibodies. The person’s blood type is determined by examining antigen-antibody reactions.
Remember, the resulting match is considered class evidence. However, if the blood does not
match, then a particular person may be omitted as a suspect. Depending on the
circumstances, blood typing may not be done at all, just DNA analysis. DNA matching could
link a suspect to a scene and it can prove a link between suspect, victim, and crime.

38
Assessing Learning

Activity 6

Name: ______________________________________________________ Rating: ____________

Course/Year/Section: ____________________________________ Date: ______________

Directions: Read each question carefully and write your final answer on the space provided.

1. Human blood types are determined by genes that follow the CODOMINANCE pattern
of inheritance. There are two dominant alleles (IA and IB) and one recessive allele (i). Write
the genotype for each person based on the description:
a. Homozygous for the “B” allele ___________
b. Heterozygous for the “A” allele ___________
c. Type O ___________
d. Type “A” and had a type “O” parent ___________
e. Blood can be donated to anybody ___________
f. Can only get blood from a type “O” donor ___________

2. Based on the descriptions, choose the best answer in the following words.
a. antibody b. homozygous c. thrombocytes d. Coagulate
e. Plasma f. Heterozygous g. Leukocytes h. Agglutination
i. Antigen j. Erythrocytes
___1. Manufactured by the white blood cells to attack any antigen that does not belong.
___2. The genes are the same.
___3. Platelets are responsible for producing fibrin to form a clot.
___4. Blood changes from a liquid to a semi-solid state.
___5. The fluid portion of the blood that carries the solid components and is made up of water
and metabolites.
___6. The genes are different.
___7. White blood cells that provide immunity and produce antibodies.
___8. Red blood cells clumping together due to antibodies in the blood.
___9. Identification proteins that are found on the surface of every red blood cell.
___10. Red blood cells contain hemoglobin and transport gases through the body.
___11. Another specific antigen found on the surface of every red blood cell.

3. On the drops in the diagram, draw an arrow to indicate the direction the blood was
moving when it hit the surface.

39
 4. List three things that a blood spatter pattern can tell us and explain.
_________________________________________________________________________________________________________
_________________________________________________________________________________________________________
_________________________________________________________________________________________________________
_________________________________________________________________________________________________________
_________________________________________________________________________________________________________
_________________________________________________________________________________________________________
_________________________________________________________________________________________________________
_

5. Describe what happens to a blood drop as you increase the height at which it was
dropped.
_________________________________________________________________________________________________________
_________________________________________________________________________________________________________
_________________________________________________________________________________________________________
_________________________________________________________________________________________________________
_________________________________________________________________________________________________________
_________________________________________________________________________________________________________
_________________________________________________________________________________________________________
_

40
UNIT VII: SEMEN
Objectives: After studying this module, you will be able to know-
1. About various body fluids.
2. About forensic aspect of these body fluids as evidence
3. About examination of the body

Nature

• produced by the combined secretions of the seminal vesicles, the seminiferous tubules,
the prostate gland, and the bulbourethral gland.
• comprised of between 1.5 and 5 ml of semen and contains 40 – 250 million spermatozoa.
• Semen analysis is usually a feature of cases of rape and sexual assault
• In crime a scene, semen can be found in vagina, rectum, or anus. Semen is a liquid form
or smear and stain
• Liquid form, and gel-like fluid

• Dry weight: 7 percent of the liquid weight

• contains sperm which has a definite morphological structure
• Morphology of the human spermatozoa is a characteristic but does not allow
identification of an individual
• Aspermic semen – when semen does not contain any sperm, could be due to disease or
vasectomy operation
• Immunological test and electrophoresis are a current technique for semen identification

Figure 1. Structure of sperm

Composition
• Made up of inorganic and organic compounds
• Important component
• Enzymes
• Blood group factors
• Choline
• Fructose
• Citric acid
• Uric acid

41
• Zinc

Acid phosphatase
• Enzyme found in semen
• Concentration is higher compared to other body fluids
• Utilized for determining the location and identification of semen through staining
• Choline gets crystal tests
• Fructose, citric acid, and zinc are not present in other body fluids could also be used

Locating the Semen

• Visual examination
• Fresh semen – colorless or pale yellow, has a characteristic smell
• Freshly dried – gives characteristic odor, stiff and rough to feel
• Ultraviolet rays
• Seminal stains give fluorescence when subjected to UV rays
• Depends on the quality and freshness of the semen

Spectromicroscopy
• May give contours and layers like formations in a thick crust of semen stain

Phosphatase method
• Filter papers are dipped in saline, garments needed to be examined is pressed against the
paper. The outline of the cloth is marked for determining the location of the semen
• Filter paper is then sprayed with acid phosphatase reagent, color spots indicate the
location and size of the stain

Examination of Semen Stain

Chemical Tests

Barberio’s Test
• Uses a saturated aqueous solution of picric acid. Spermine picrate crystals with
characteristic structure, separate out.

Florence test
• Uses iodine in potassium iodide solution. Characteristic crystals of choline iodide are
formed.

Acid phosphatase test

• Takes advantage of the action of phosphatase. The presence of phosphatase in semen
removes Naphthol from calcium naphthyl phosphate. Naphthol reacts with a diazonium
compound to give colored products. Anthraquinone-1 diazonium chloride is often used
in the test
• Development of red-purple color indicates semen; the technique is extremely sensitive
and several years old stains can thus be detected.

Morphology of spermatozoon
The sperms may be identified with or without staining. But staining helps:

42
• Ordinarily double staining with hematoxylin and eosin is done. In this way, the nose is
stained in pink and the rest of the head is stained purple. This differential staining
distinguishes the sperm from any other organism. Head alone can identify the stain.
• Epithelial cells and leucocytes in excess mask the spermatozoon the difficulty is overcome
by staining the spermatozoon with malachite green and eosin. The interference is
considerably reduced.
Biological Tests
• Uses precipitin that reacts with anti-human semen serum and specific blood group
antisera
• Determines human semen and blood group of secretors respectively

Individualization
• Uses DNA profiling technique, accurate even contamination of the vaginal fluid is present
or other sources are available

Age of seminal stains

• Difficult to determine since highly dependent on the environmental condition
• Gel-like mass after ejaculation
• Liquefies after standing
• Opaque white mass with rough feel after drying
• Color changes to pale yellow and brown after two to three weeks

• Sperms in fresh semen are complete with head and tail. On drying and with the passage
of time it begins to disintegrate, the tail separates out from the head. The head is also
affected. The sharp division in differential staining slowly appears.

Survival of sperms
• Sperm is motile for a few hours under normal condition
• When frozen, may continue to live for a long period of time

• Warm climate – sperm is easily destroyed

• Cool climate – dead sperms may be identified even after months
• Vaginal swabs – sperm can be detected five to ten days
• Dead persons – can be detected in unputrefied bodies even after a month, sperms are
destroyed in putrefied bodies

43
UNIT VIII. HAIR AND FIBER
Learning Objectives
At the end of the unit, I can:
1. Identify the various parts of the hair.
2. Describe variations in the structure of the medulla, cortex, and cuticle.
3. Distinguish between human and nonhuman animal hair.
4. Determine if two examples of hair are likely to be from the same person.
5. Explain how hair can be used in a forensic investigation.
6. Distinguish hairs from individuals belonging to the broad racial categories.
7. Identify and describe common weave patterns of textile samples.
8. Compare and contrast various types of fibers through physical and chemical analysis.
9. Describe the principal characteristics of common fibers used
10. in their identification.
11. Apply forensic science techniques to analyze fibers.

A. Hair

• Hair analysis is important during the investigation of a crime. Hair is trace evidence
during an investigation. An example of a case where hair was found to be useful is the
murder of Duchesse de Praeslin in Paris in 1847
• The use of the comparison microscope to perform side-by-side analysis of hairs
collected from a crime scene and hairs from a suspect or victim
• Function of Hair
o Regulate body temperature through insulation
o Lessen friction
o Protection against sunlight
o Act as a sense organ
• If the temperature is low, muscles pull the hair upright thereby creating a pocket of trap
air, this trapped air provides a warm layer next to the skin. And if the temperature is
high, the muscle relaxes, and the hair becomes flattened releasing the trapped hair

Structure of Hair

Two parts
• Follicle
• Shaft

Follicle
• Consist of a network of blood vessels for nutrition
supply for hair growth – papilla
• Bulb – surrounds the papilla
• Sebaceous gland – secretes oil that maintains the hair
conditioned
• Erector muscle – makes the hair stand up
• Nerve cells – for response in the environment
Figure 1. Structure of the hair

44
Shaft
• Contains protein keratin
• Keratin makes the hair strong and flexible
• Made up of amino acids that form a helical and spiral shape

Three layers of the shaft

1. inner medulla
2. cortex
3. outer cuticle

Cuticle
• transparent outer layer
• has scales that overlap with each other
• points from the proximal end to the distal end of the hair
• the direction of the scales shows the region of the older and newer hair
• can be used in determining the presence of toxin and drugs at a specific time frame
• human hair has cuticle scales also known as imbricate
Cortex
• the largest part of the hair shaft
• contains the pigment granules melanin
• varies distribution from person to person when viewed under a microscope

Medulla
• center of the hair
• can be hollow or filled with cells
• can contain pigments or not
• Investigators classify hair depending on the medulla
Table 1. Types of Medulla

Types of Hair
• vary in shape, length, diameter, texture, and color
• the cross-section can be circular, triangular, irregular and flattened can be coarse and
fine
• Hair varies from different parts of the body

45
Types of human body hair
1. head hair
2. eyebrows and eyelashes
3. beard and mustache hair
4. auxiliary or body hair
5. pubic hair

Figure 2. Types of Human Hair

• can be distinguished through physical features and cross-section

Life cycle
Stages
1. Anagen stage - lasts 1000 days, active growth, follicles are rapidly dividing and supply
nutrients to the cell
2. catagen stage - turning gray
3. telogen stage - dormant and easily lost

Treated hair
• bleaching removes pigment
• brittle and shows sharp demarcation
• dyeing hair changes the color of the shaft which can be seen in the cuticle and cortex
• if an entire hair is recovered from a crime scene, the region near the root of the hair is
colored naturally, hence this can be used in the investigation
Racial difference
• there are physical characteristics that are associated in different racial groups but may
not apply to all individuals

Table 2. Racial

46
Animal Hair and Human Hair
• animal and human hair differs in pattern and pigmentation, medulla and cuticle type
• Ovoids - solid mass of pigments in animals
• Medulla in animals is larger than in humans
• medullary index - the ratio of the diameter of the medulla to the diameter of a hair

Hair in an investigation
• Lockard’s exchange principle - when two objects are in contact, transfer of materials
occurs.
• reason for the use of trace evidence in an investigation collection
• plucking, shaking and scraping surfaces placing tape over surfaces vacuum
• an investigator must prevent cross-contamination
• comparison of the collected hair sample to the victim or suspect
Microscopy
• hair is studied both macroscopically and microscopically
• typical magnification is 40x
• characteristic to be studied
o medulla pattern
o cortex pigmentation
Microscopes
1. phase-contrast microscope with a special condenser to show more contrast
2. Fluorescence microscope - uses dyes
3. Electron microscope
Substances in the hair shaft
• toxins can be detected in the hair like arsenic, lead, and drugs
• the keratin of the hair is dissolved
• hair can be used to establish a timeline for toxin exposure since hair does not easily
decompose
• human hair grows at the rate of 1. 3 cm per month, thus can be analyzed in the section
for the specific toxin
• Neutron activation analysis - can be used to identify 14 different elements in a single
strand of hair
Testing for hair follicle
• follicular tag - when the entire follicle is present
• Blood proteins can be isolated and analyzed
• DNA analysis can also be done

B. Fibers

• Fibers are used to create a link between the crime and the suspect
• an example of trace evidence
Sources
• carpets, clothing, linens, furniture, insulation, or rope.
Type of transfer
• direct transfer - transfer of fiber from the victim to suspect or suspect to the victim
• secondary transfer - victim has fibers on his person that he picked up and then
transferred to a suspect
• Clothes shed fibers

47
• Criminals wear gloves to prevent this but unaware of very small fibers which are
important evidence
• the early collection is crucial
How Forensic Scientists Use Fibers
• value depends on the uniqueness of the fiber since other fibers are also present in a
crime scene
• a forensic scientist should look for the following
1. Type of fiber
2. Fiber color
3. A number of fibers found.
4. Where the fiber was found
5. Textile the fiber originated from.
6. Multiple fiber transfers.
7. Type of crime committed
8. The time between crime and discovery of fiber

Sampling and Testing

• Sampling
o special vacuums
o sticky tape
o forceps
• important to record where you found the fiber
• identified fiber should be matched with the suspect source such as home and car

Methods to analyze
• polarizing light microscope
• infrared spectroscopy
• burning or dissolving in liquid to identify the fiber is an option if collected in bulk

Fiber and Textile Evidence

• textile - things like clothes, carpets, and upholstery
• yarns - use to construct textiles by weaving or intertwining single units which are made
up of fibers spun together

Classification
1. natural
2. synthetic

• Important to reveal the environmental information of the suspect

Natural Fibers

Source:
1. Animal
• come from hair, fur, and webbing, made up of proteins
• used in clothing, carpets, curtains, and bedding

2. Plant
• specialized plant cells

48
• group depending on which part of the plant they came from
• diverse: can be thick or thin, smooth, fine, flexible or amorphous
• Share a common feature: the presence of cellulose
• Cellulose is a polymer of glucose molecules
• Glucose is a carbohydrate that allows the scientist to distinguish plant fibers from
animal fibers

• Seed fibers- cotton can be found inside a seedpod when dyed and woven, it can be used
for clothing and household textiles

• Fruit fiber- coarse and can be obtained coconut, cells from coir fibers are narrow and
has thick walls made up of cellulose

• Stem fibers- Hemp, jute, and flax are all produced from the thick region of plant stems,
unlike cotton, stem grow in bundles that can reach up to six feet
o processing includes beating and rolling the stem until the fibers separate
o Flax is an example of stem fiber that can be found in textile linen
o Linen is unique because it is highly crystalline, so it is a dense, strong fiber that
resists rot and light damage

• Leaf fibers - Manila is a fiber extracted from the leaves of abaca, a relative of the banana
tree
o fiber bundles are extracted from the surface of leaves
o can be found in ropes, twines, and netting

3. Mineral fiber
• neither proteins nor cellulose
• fiberglass is an example
• asbestos, a mineral that can be found in different types of rocks
o has a crystalline structure
o long and thin fibers
o uses include pipe coverings, brake linings, ceiling tiles, floor tiles, fire-
resistant work clothes, shingles, home siding, and insulation for building
materials
Synthetic fibers
• Categorized into regenerated fibers and polymers
• Polymers – produced by joining monomers
Regenerated fibers
• derived from cellulose
• rayon is the most common type
• cellulose + acetate produces Celanese® that is used in carpets
• cellulose combine with three acetate unit forms polyamide-nylon
Synthetic polymer fibers
• originates from petroleum products and does not contain cellulose
• have no definite size and shape
• can be distinguished using a polarizing microscope or infrared spectroscopy

49
• Polyester: A very common synthetic fiber, polyester represents a very large group of
fibers with a common chemical makeup. It is found in polar fleece, wrinkle-resistant
pants, and is also added to many natural fibers to provide additional strength.
• Nylon: Like polyester except that it can be easily broken down bt acid
• Acrylic: found in artificial wool, has a light and fluffy feel
• Olefins: found in high-performance clothing such as thermal socks and carpets, quick-
drying and resistant to wear
• Yarns are woven, with different patterns, into textiles

Weaving pattern
Table 3. Weaving
Pattern

50
Assessing Learning

Activity 7

Name: ______________________________________________________ Rating: ____________

Course/Year/Section: ____________________________________ Date: ______________

1. Locard’s exchange principle implies all of the following except

a) Fibers can be transferred from one person to another.
b) Blood spatter can be used to identify blood type.
c) Cat hair can be transferred to your pants.
d) Soil samples can be carried from the yard into your home.

2. Transfer evidence can include all the following except

a) the victim’s own blood gushing from a wound
b) hair that was transferred to a hairbrush
c) the blood of the victim found on a suspect
d) a footprint

3. The reason it is important to separate the witnesses at the crime scene is to

a) prevent contamination of the evidence
b) prevent fighting among the witnesses
c) prevent the witnesses from talking to each other
d) protect them from the perpetrator

4. Correct collection of evidence requires which of the following?

a) documenting the location where the evidence was found
b) correct packaging of evidence
c) maintaining proper chain of custody
d) all the above

5. A crime-scene sketch should include all the following except

a) a scale of distance
b) date and location of the crime scene
c) a north heading on the diagram
d) the type of search pattern used to collect the evidence Short Answer

6. Distinguish between circumstantial evidence and direct evidence and provide an example
of each type.

51
UNIT IX. GUNPOWDER & EXPLOSIVES
Overview
In the investigation of crimes involving the use of firearms, law enforcement
officers need to know whether a suspect has fired a gun or not and the gunshot range
or the approx. the distance of the shooter to the victim. When a firearm is discharged
unburned particles of the gunpowder may escape thru the breech of the gun and
deposit on the hands of the shooter, on the clothing of the victim and some may
remain in the barrel. These gunpowder residues can be detected by chemical
examination.
In connection with suspected bombings, the lab is requested to detect traces of
explosives that may be found in the remains of parts of a bomb. Explosives like
improvised and manufactured dynamite used in illegal fishing, are also examined.
Learning Objectives
At the end of the unit I am able to:
1. Define relevant terms.
2. Identify the chemical and physical properties of gun powder and explosives.
Lesson Proper
In the investigation of crimes involving the use of firearms, the three most
important problems may arise:
1. Determination of whether or not a person has fired a gun with his bare hands
within a pertinent period of time.
2. Determination of the probable gunshot range, which is the distance the firearm
was held from the body of the victim at the time of discharge.
3. Determination of the approximate time of firing of the gun or the approximate
date of last discharge.
Two Kinds of Gunpowder
1. Blackpowder – It is consists of an intimate mixture of charcoal (Carbon -15%),
(Sulfur – 10%), and (KNO3 or NaNO3 – 75%).
2KNO3 + 3C + S → 3CO2 + K2S + N2
2. Smokeless powder - It is consists of cellulose nitrate or glyceryl nitrate combined
with cellulose nitrate and some stabilizers. When exploded the following reactions
occur:
C12H14O4(NO3)6 → 9CO + 3N2 +7H2O + 3CO2
cellulose nitrate

52
 4C3H5(NO3) 3 → 12CO2 +10H2O + 6N2 + O2
glyceryl nitrate
Possible locations of gunpowder residue
1. Residue of the barrel of the gun.
2. In or around the wound.
3. On the clothing of the person fired upon at close range.
4. On the exposed surface of the hand of the person firing the gun.
Factors that affect the presence and amount of gunpowder residues
1. Type and caliber of the ammunition.
2. Length of the barrel of the gun.
3. The distance of the muzzle of the gun from the target
4. Humidity
5. Wind velocity and direction.
6. The direction of the firing.
Paraffin test (Dermal Nitrate test or Diphenylamine test or Lunge’s test) – a test to
determine whether a person fired a gun or not with his bare hands. The basis of this
test is the presence of nitrates in the gunpowder residue. Warm melted-paraffin wax
applied on the hands of the shooter will cause the pores of the skin to open and exude
the particles of the gunpowder residue. These particles are collected by the paraffin
cast and will appear as blue specks when the diphenylamine reagent is added to the
cast. The specks are generally located in the area of the thumb and forefingers.
Positive Result – Deep blue specks develop when nitrates come in contact with
diphenylamine reagent.
Note: The result of this test is not conclusive because it does not always show that
nitrates or nitrites were necessary results of the discharge of firearms. Other
substances yield the same positive reaction for nitrates or nitrites, such as explosives,
fireworks, pharmaceuticals, and leguminous plants such as peas, beans, and alfalfa.
(People v. Baconguis, 417 SCRA 66, December 2, 2003).
Factors to be Considered in the Interpretation of Paraffin Test Result:
1. Time of reaction
2. Number of specks
3. Location of specks
4. Character of specks
Other Sources of Nitrates:
1. Fertilizers
2. Urine

53
3. Explosives
4. Cosmetics
5. Tobacco
6. Detergents
7. Firecrackers
Possibilities That a Person Maybe Found Negative Even If He Actually Fired a Gun:
1. Use of automatic pistol
2. Direction of wind
3. Wind velocity
4. Excessive perspiration
5. Use of gloves
6. Knowledge of chemicals that will remove the nitrates

Determination of the Probable Gunshot Range:

The clothing of the victim with bullet perforations should be submitted for possible
gunshot range. It is only possible to determine the probable gunshot range if the gun
was fired from a distance of zero (contact firing) to 36 inches (3 feet) based on the
distribution of powder patterns or the spread of a shot pattern.
Three Zones of Distance
1. Those in which the muzzle of the gun was held directly in contact with the body
or practically so. (contact wounds).
Contact wounds – Damage (on cloth) is due more to the flame and the muzzle blast
than to the penetration of the bullet. Describe by the following;
a. The gaping hole where fabric is badly torn,
b. The blackened area surrounding the bullet hole,
c. Singeing of the fibers at the entrance, and
d. Presence of partially burned powder residues around the entrance hole.
2. Those in which the muzzle of the gun was held from about 2 inches to 36 inches
away.
Distance from which the firearm was discharged Characteristic Pattern
4. Those in which the muzzle of the gun was held from 36 inches or more

54
 Distance from which the Characteristic Pattern
firearm was discharge

0 to 2 inches Gaping hole, smudging, singeing, tattooing and

burning. Smoke & soot is deposited around the hole of
entrance producing a dirty, grimy appearance.
Individual specks of tattooing around the hole are
visible with the naked eye.

2 inches to 8 inches Smudging

8 inches to 36 inches Partially burned and unburned powder particles are

driven into the surface around the gunshot hole
producing a black coarsely peppered pattern called
tattooing.

Beyond 36 inches Evidence of powder tattooing is seldom present.

Nitrate particles may be present.

Determination of the Probable Time The Gun Has Been Fired

1.Rust: Formation of rust inside the barrel after a gun has been fired is a good
indication for the determination of the approximate time the gun has been fired.
a. If a gun has not been fired at all, no rust can be detected inside the barrel of the gun.
b. If a gun has been fired, iron salts are formed and are found inside the barrel. These
iron salts are soon oxidized resulting in the formation of rust.
2. Presence of nitrite (NO2) – firearms could have been fired recently. Gives blue color
in paraffin test results.
3. Presence of nitrate (NO3) – firearms could have been fired but not recently. Gives
a yellow-green color in paraffin test results.
Types of Explosives
1. Mechanical – such as those caused by the expansion of gas-producing high pressure
beyond the capacity of the container.
2. Atomic – resulting from atomic transformations.
3. Chemical – produced thru the extremely rapid transformations of the unstable
substances accompanied by the formation of heat.
Classification of Explosives

55
 The speed of the chemical reaction or detonation of the explosives determines
the classification of explosives as low, initiating, or high explosives.
1. Low explosives are low burning and are used mainly as propellants, like black
powder and smokeless powder. Atomic explosive results from atomic
transformations.
2. Primary or initiating explosives are extremely sensitive to detonation by heat,
shock friction, and impact. They detonate without burning, like lead oxide and
mercury fulminate.
3. High explosives detonate under the influence of shock of the explosion of a primary
explosive. Examples are the following:
a. Ammonium nitrate – the cheapest and most readily available salt of nitric acid.
b. TNT (trinitrotoluene) – the most widely used explosive.
c. Nitroglycerine – most widely used commercial explosive.
d. RDX – considered one of the most important military explosives used today.
e. C4 – a plastic explosive that is white and dough-like in texture.
f. Chloroacetophenone – CN is the principal component in the filler used in tear gas
solutions.
4. Fire Bombs – Molotov cocktail which is an incendiary device, acids mixed with
gasoline, alcohol, and gasoline, etc.
References
Melcon S. Lapina, MSCrim, Forensic Chemistry, and Toxicology, [Powerpoint slides]
Aries Gallandez, Forensic Chemistry, International School of Asia and the Pacific,
[Document ]

56
Assessing Learning

Activity 8

Name: ______________________________________________________ Rating: ____________

Course/Year/Section: ____________________________________ Date: ______________

Directions: Shade the letter of your corresponding answer in the answer sheet below.
1. What is the maximum distance that is determined by firing?
a. 36 inches c. 12 inches
b. 43 inches d. 25 inches
2. To determine whether a wound is a contact or not, there must be:
a. burning c. nitrates
b. gaping hole around the wound d. none of the above
3. Test that determines whether a person fired a gun with his hand is:
a. diphenylamine c. precipitin
b. agglutinin d. none of these
4. Gunpowder residues are also known as:
a. nitrates c. nitric acid
b. hydrochloric acid d. none of these
5. It ________ consists of a mixture of carbon, sulfur and potassium nitrate.
a. black powder c. nitrates
b. smokeless d. mercury
6. It ________ consist of a mixture of nitrocellulose and nitroglycerine.
a. smokeless c. nitrates
b. black powder d. none of these
7. Mechanical, atomic and chemical are classified as:
a. explosives c. Molotov bomb
b. pill box d. ammunition
8. Low explosives, primary or initiating and high explosives are classified as:
a. explosives c. destructive
b. flammable d. none of these
9. The basis of paraffin-diphenylamine test is the presence of:
a. nitrates c. acid
b. explosives d. all of these
10. A more specific test employed by the FBI laboratory in paraffin test is:
a. neutron activation analysis c. atomic absorption
b. spectrographic method d. none of these
11. The diphenylamine reagents are applied to cotton swabbing of the bare hand
chambers of the gun to determine if the firearms was:
a. fired c. swabbed
b. cleaned d. dried

57
12. When a firearm/gun is discharge-residue may also be deposited on the clothing
of the victim.
a. gunpowder c. diphenylamine
b. chlorate d. none of these
13. Main factor considered in determining the approximate distance of the gunman
to the victims is:
a. the size and density of the pattern of gunpowder residues
b. the distance
c. the place
d. the time
14. Singeing, smudging, and tattooing are called characteristics:
a. patterns c. metallic fragments
b. particles d. residues
15. Bombs should be handled only by qualified _______, therefore, do not handle
bombs unless qualified.
a. expert’s c. technicians
b. investigator d. laymen

58
UNIT X CHEMISTRY OF COMBUSTION AND ARSON
Overview
This unit focuses on the special topic underlying the principles of arson which is
combustion. As a forensic science the unit focuses on an exploration of the
fundamentals of combustion, a specialized type of oxidation–reduction that spans
candle flames to powerful and destructive explosions. The important differences
between a fire and a bomb are the speed at which the combustion occurs and the
degree to which it is confined. In this unit, you will cover and recognize the
fundamental principles that underlie these types of forensic cases and evidence and
discuss the physical evidence associated with fires.
Learning Objectives
In this unit, I am able to:
1. Identify the fundamental principles of combustion
2. Discuss the physical evidences associated with fire
3. Identify the sources and types of fire source.
4. Identify the basic principles to be observed under a fire investigation
Lesson Proper
Combustion is an oxidative decomposition in which oxygen (the oxidant) oxidizes a
fuel. The different manifestations of combustion, ranging from a gentle candle flame
to a violent military explosive, are part of a continuum that includes the propellants
used in firearms. The terms used to describe this decomposition depends on
characteristics such as the speed of the reaction. For example, in the detonation of an
explosive, the reaction front moves faster than the speed of sound (~ 741 mph, or 331
m/s). If this same reaction occurs at less than the speed of sound, it is called
deflagration . This term can also be applied to what we mean by the term “burning,”
in which the flame speed is less than the speed of sound. As the flame front’s speed
increases, oxygen from the atmosphere is incapable of sustaining it, and additional
oxygen must come from another source such as potassium nitrate ( KNO3 ). The fuel
must also change as the energy derived from it increases.
Combustion is an exothermic reaction in which reactants are converted to principally
gaseous products. Because the reaction is exothermic, the product gases heat
up and expand, and in turn can be harnessed to do work. In a fire, such as arson, this
expansion generates plumes with predictable behaviors that leave distinctive
markings

59
at the crime scene.
Combustion requires reactants and enough energy to exceed the energy of activation
(Ea) required to initiate the reaction. The reaction profile illustrates the exothermic
nature of a combustion reaction, as well as the need for
enough energy to initiate it. Once initiated, enough energy is produced to supply the
necessary Ea to sustain the reaction until one of the reactants is exhausted. With a
simple flame, the fuel is exhausted first, since the oxidant is atmospheric oxygen.
When chemical oxidants are employed, either fuel or oxidant may be the limiting
reagent.
You could be familiar with the fire triangle, which is one way of summarizing the
requirements for a combustion reaction. Such a triangle is divided into three regions,
identified as fuel, oxidant, and heat (the last of which supplies Ea ). Building on the
concept of the triangle, we will consider the requirements for combustion to be
1. fuel and oxidant in appropriate quantities and concentrations,
2. a source of Ea, and
3. sufficient contact time for the energy source to initiate the reaction.
The absence of any one factor prevents combustion.
The first requirement, fuel and oxidant in proper proportions, illustrates key points
and unmasks common misconceptions. Wood does not burn; rather, what burns are
the vapors emanating from heated wood. Gasoline in a can will not explode because
the proper fuel–air mixture does not exist. A cigarette tossed into a pool of gasoline
usually smothers before it has a chance to ignite the vapor above it. Similarly, the
Hollywood staple of exploding gas tanks in cars is more fiction than fact. Rapid
burning can occur, but only when the gas tank is ruptured, the contents leak and
vaporize, and the proper air–fuel vapor mixture is created at the same time and place
as a source of ignition that stays in contact long enough to spark the reaction.
Aspects of Combustion
Combustion, with rare exceptions, is a complex chemical process involving many
steps that depend on the properties of the combustible substance. It is initiated by
external factors such as heat, light, and sparks. The reaction sets in as the mixture of
combustibles attain the ignition temperature. The combustion spreads from the
ignition source to the adjacent layer of gas mixture; in turn, each point of the burning
layer serves as an ignition source for the next adjacent layer, and so on. Combustion
terminates when equilibrium is achieved between the total heat energies of the
reactants and the total heat energies of the products. Most reactions terminate when
what is called thermal equilibrium has been attained, especially when the energy of
the reactants equals the energy of the products.

60
Kinetics of Combustion
Combustion does not involve simple single-step collisions between a fuel molecule
and an oxidant. Rather, the reactions that occur during combustion are based on free
radicals. In a free-radical 2 mechanism, three generic steps take place: Initiation , in
which the first free radicals are formed; Propagation , in which reactions among
radicals produce more radicals; Termination , which results from the combination of
two free radicals to form a neutral species.
Fires and Flames
A common type of fire or flame is a laminar flame , meaning that the flame has
definable layers or regions. Within the laminar category, there are two familiar
examples: a candle and a Bunsen burner, shown in Figure In a Bunsen burner,
gaseous methane is mixed with air and injected into the combustion zone. Hot
gaseous products diffuse away while atmospheric oxygen diffuses inward, creating a
second region in which the fuel–air mixture is no longer in the same proportions as
in the premix zone. In a candle, fuel is vaporized and diffuses upward as oxygen
diffuses inward. The regions in the candle flame are defined by temperature.
Pyrolyzed waxes form particulates (soot) that grow and rise. Partial oxidation occurs,
and any residual soot diffuses upward and outward as smoke. In both cases, the flame
is self-sustaining once ignited and burns until the fuel is exhausted. A visible
combustion wave is created if the fuel and oxidant gases are stationary, A flame front
is visible when, for example, a large puddle of gasoline is ignited at one end and the
flames propagate across the surface much as waves on a pond propagate from the
point where a stone is dropped into the water. Typical flame velocities for a mixture
of methane and air are in the range of approximately 15 to 45 cm/sec, depending on
factors such as the fuel–air ratio.4 In a candle or Bunsen burner, the wave front is
stationary because the gases are moving into and out of the combustion zone. The
combustion wave remains stationary because the unburned gas molecules are
moving away at a rate equal to the burn rate. In simple combustion (deflagration), the
combustion wave never exceeds the speed of sound.

Arson and Fire Investigation

Arson Star City Incident
After years of providing family-friendly entertainment and Christmas cheer, the
popular amusement park Star City went up in flames.
According to Star City’s management, it will be closed this Christmas season after 60
percent of the compound perished from the fire that lasted for hours. News reports
cited Atty. Rudolph Steve Juralbal, vice president for legal affairs of Star City’s owner,
Elizalde Group of Companies, as saying: “We’re targeting reopening about October

61
[2020]. It’s necessary because importation of rides takes time and the set-up of the
park also.”
The Bureau of Fire Protection (BFP) inferred that arson could have caused the fire
that hit Star City. While BFP initially found that the fire started from the stockroom, it
noted that different parts of Star City seemed to have started burning at the same
time. Moreover, Lisa Macuja, wife of Fred Elizalde of the Elizalde Group of Companies,
reported to the BFP a social media post stating that, “Star City will die.”
However, Arson was ruled out in the Star City incident. BFP spokesperson Gerrandie
Agonos told CNN Philippines that the ballast of a fluorescent lamp in the compound
overheated and started the fire last October 2. This debunked the arson theory earlier
proposed by Pasay City Fire marshal Supt. Paul Pili, who noted the presence of
gasoline in one of the indoor rides and cotton brought into the park prior to the
incident.
Destructive arson contemplates the malicious burning of private and public
structures, hotels, buildings, edifices, trains, vessels, aircraft, factories and other
military, government or commercial establishments by any person or group of
persons.
It is punishable by reclusion perpetua to death so as to effectively discourage and
deter the commission of this dastardly crime. In imposing these severe penalties, the
following should be considered: (a) extreme danger to human lives exposed by the
malicious burning of these structures; (b) danger to property resulting from the fire;
(c) the fact that it is normally difficult to adopt precautions against its commission;
(e) the difficulty in pinpointing the perpetrators; and (f) the greater impact on the
social, economic, security and political fabric of the nation.
Destructive arson is characterized as a heinous crime for being a grievous, odious and
hateful offense and, which by reason of its inherent or manifest wickedness,
viciousness, atrocity and perversity, is repugnant and outrageous to the common
standards and norms of decency and morality in a just, civilized and ordered society.
Meanwhile, simple arson contemplates the malicious burning of private and public
structures, regardless of their size, and not otherwise classified in relation to
destructive arson. These include houses, dwellings, government buildings, farms,
mills, plantations, railways, bus stations, airports, wharves and other industrial
establishments.
Unlike in the relevant provisions on destructive arson, the law tempers the penalty to
be meted to persons who have committed simple arson. This separate classification
recognizes the need to lessen the severity of punishment commensurate to the act or
acts committed, depending on the particular facts and circumstances of each case.

62
The penalty for simple arson shall be imposed in its maximum period if any of the
following special aggravating circumstances were present: (a) if committed with
intent to gain; (b) if committed for the benefit of another; (c) if the offender is
motivated by spite or hatred toward the owner or occupant of the property burned;
and (d) if committed by a syndicate or group of three or more persons. Moreover, if
by reason, or on occasion of simple arson death results, the penalty of reclusion
perpetua to death shall be imposed.
Based on the foregoing definitions, determining whether the crime is destructive or
simple arson would depend on the kind, character, and location of the property
burned, regardless of the value of the damage caused. Likewise, while intent may be
an ingredient of the crime of arson, it may be inferred from the acts of accused. Thus,
when the offender is shown to have deliberately set fire to a building, the prosecution
is not bound to provide further evidence of his wrongful intent. If there is an
eyewitness to the crime, he can give in detail the acts of accused. Furthermore, the
prosecution may describe the theatre of the crime and the conditions and
circumstances surrounding it.
Source: https://business.inquirer.net/280440/explaining arson#ixzz6TLmf35cu
Intentionally set fires, or arson, usually involve an accelerant of some type, as well as
an incendiary device used to ignite it, and these two components create the physical
evidence forensic chemists work with. Arson fires are also referred to as incendiary
fires. For fires set in homes, a bedroom was the most common point of origin; fires
set in public buildings were usually started in bathrooms. From a law enforcement
point of view, arson is a difficult crime to clear; in 2008, only 18% of arson cases were
cleared by arrest or other means. Nearly half of those arrested were under 18, and
3% were under 10 years of age.
One of the challenges of fire investigation is the classification of a fire as
natural,accidental, or incendiary (arson). investigators utilize evidence at the scene,
as well as forensic analysis, to make such determinations. One of the most important
pieces of information required in making a determination of arson is the location of
the point or points of origin of the fire. Multiple points of origin are strongly indicative
of an intentionally set fire, whereas a point of origin at an electrical outlet suggests an
accidental fire.
Accelerants and Incendiary Devices
An accelerant is anything that is intentionally placed at a scene to start and sustain a
fire. Accelerants can be solids (wood, paper, road flares), liquids (gasoline), or gases
(natural gas, propane). Although a gaseous accelerant will not leave chemical
residues, there will always be physical evidence related to the container used to hold
or deliver it. For example, if bottled propane is used to start a fire, the bottle will
remain even though the propane will not. As with any type of combustion, fuel,

63
oxidant, and a source of ignition are required, and the forensic chemist will be
concerned with two of these three: the fuel, such as an accelerant, and the incendiary
device. The latter can be as simple as a match, a candle, or a smoldering cigarette.

As the name implies, petroleum distillates are extracted from crude oil by distillation.
It is the most useful because of the parallels between distillation and gas
chromatography. Crude oil is introduced into the cracking tower and heated to about
350°C, volatilizing much of its content. The vapors rise, cool, and condense,
whereupon they are collected on plates and removed. The heavier fractions, such as
diesel and kerosene, are collected lower in the tower than are gasolines and fuel
gases.
The separation is not complete, and each fraction collected consists of a mixture of
hydrocarbon components with similar boiling points. This separation is re-created on
a small scale when samples are analyzed by gas chromatography, which separates
components on the basis of their volatility and preferential partitioning on a solid
phase.
Forensic Analysis of Fire Debris
The most frequently encountered accelerants are readily available petroleum
distillates such as gasoline, jet fuel, and kerosene. ASTM has published a classification
for ignitable liquids that is widely used as part of the forensic analysis of fire
debris.Each class is further subdivided into three groups based on carbon chain
length, which correlates with volatility. Fire debris evidence is collected in coated
paint cans of various sizes with a predrilled hole sealed with a septum. The debris is
placed loosely in the can, leaving plenty of headspace above. As with all paint cans,
the can is closed by pounding the lid into the metal lip. This creates an airtight seal
that traps vapors, including any residual volatile accelerants.
References
Akhavan, J. “Thermochemistry of Explosives.” Chap. 5 in The Chemistry of Explosives
. Cambridge, U.K.: Royal Society of Chemistry, 1998.
Almirall, J. R., and K. G. Furton, eds. Analysis and Interpretation of Fire Scene Evidence
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