SCHOOL OF NATURAL SCIENCES

General Luna Road, Baguio City Philippines 2600

Telefax No.: (074) 442-3071 Website: www.ubaguio.edu E-mail Address: [email protected]

A Learning Module in

FORENSIC CHEMISTRY
AND TOXICOLOGY

https://www.waynesburg.edu/
Introduction
I. Course Code and Title: FORCHM2 (Forensic Chemistry and Toxicology)
Course Descrip.tion and Information: This is a 5-unit course with 3 units lecture and 2 units laboratory.
Forensic Chemistry deals with application of chemical principles in the solution of problems that arise in
connection with the administration of justice. It is chemistry applied in the elucidation of legal problems.

II. Requirements of the Course:

1. Regular Attendance to classes: you must attend online classes and live quizzes regularly by
logging in to our scheduled online activities. Online lectures will be done through Canvas and/or
facebook live. Assessments shall be given through Quizziz, Pear Deck, Canvas and/or Google
forms. For offline students, your attendance will be monitored through your responses to text
information and through timely correspondence. Offline students will also be given quizzes and
activities through phone calls and text messages.
2. Submission of required activities: All required activities (assignments, research work, and
laboratory illustrations) should be submitted on or before the given deadline. Deadlines will be
posted by the teacher in the google classroom and messenger group chat. It will also be texted to
offline students. Learning output for online students, submit to the teacher’s email address that
will be given during the class orientation; For offline students, submit via mail or express courier
(“padala”) addressed to: Instructor’s name, School of Natural Sciences, University of Baguio,
Baguio City.
3. Study/Learning Guidelines:
a. Manage your time properly. As students of higher education (College), you are expected to be
more responsible in paying attention to course schedules, requirements, and deadlines.
Schedule how you will accomplish all the requirements in all your enrolled courses (reading
the modules, reading on research/ enhancement questions, doing assignments and laboratory
illustrations) and focus your attention when doing your tasks.
b. Observe proper conduct. Despite this online mode of learning, you must still maintain
appropriate school behavior at all times. All standards of student conduct outlined in the
University of Baguio Student Handbook remain in full effect during this time of distance
learning. Be honest in answering your quizzes and exams. Work independently in doing your
tasks and assignments.
c. Maintain a performance of high standards. Give your best in accomplishing all the assigned
tasks. Do not be complacent with just a 70% passing cut score.
d. Communicate properly. Promptly respond to notifications by regularly visiting our google
classroom and messenger group chat. If you have confusions or queries in any part of this
module, I am here to guide you through. Send your academic concerns using same online
platforms. For offline students, text messages and mobile calls are welcome during scheduled
hours of the day and week. Be guided by this schedule when communicating:
 Respect private hours. I do not always open my laptop/email/messenger 24/7. Send
your queries and/or concerns during regular office hours. For concerns that need
immediate attention, send through mobile text.
e. Show mutual support. Support one another. Let us all be responsible and supportive in making
this new learning process more effective.
f. Live lecture/Video conferencing guidelines:
f.1 Be punctual. Live lectures/Video conferences will be scheduled during the official class
period/time of this course. Log in to the platform at least 5-10 minutes before the class
period. Prepare your learning materials such as this module, pens, papers, etc.
Attendance will be checked during the lecture/video conference.
f.2 Maintain professionalism.
 - Wear appropriate clothing and set your gadget in an appropriate area. You may be
asked to turn on your video/camera at any time during the lecture.
- Log in using your UB gmail account. Unidentified names like nicknames, phone
models, etc. will not be allowed in the video conference.
- Mute your microphone as soon as you log in to the platform to avoid any excess
background noise. Unmute your microphone when instructed to do so.
- Respect privacy. Do not take a screenshot, picture, snapchat, etc. of your teacher or
fellow students, nor make any unnecessary audio or video recordings.
f.3 Remain focused and engaged. Do not be distracted by your gadget. Keep your
videoconference

Rubrics
Rubrics for the evaluation of Essay question activities and Questions for research
CRITERIA HIGHEST Outstanding Satisfactory Poor
SCORE

1. Content and Focus 7 Sharp, distinct & Apparent point made No apparent
-Awareness about the substantial controlling about a specific topic point and no to
specific topic point about a specific or question with minimal
-Presence of relevant ideas question or topic with sufficient awareness evidence of
thru tacts, examples, details, evident awareness and knowledge. (4 awareness and
opinions and explanation and knowledge. (7 points) knowledge. (1
points) point)
2. Organization 5 Sophisticated Functional Confused or
-Order developed and arrangement of arrangement of inconsistent
sustained with in the content with evident content that sustains arrangement no
paragraph and/or subtle a logical order w/ logical order or
transition (5 points) some evidence of evidence of
transition (3 points) transition. (1
point)
3. Style and Conventions 3 Good grammar, Sufficient grammar Incorrect
-Choice, use and spelling and sentence and minor spelling grammar and
arrangement of words formation throughout errors and sentence major spelling
-Grammar, mechanics, the paragraph. (3 formation (1 point) errors
spelling, usage & sentence points) throughout the
formation paragraph. (0
point)

Rubrics for the evaluation of Illustrations and Diagrams

CRITERIA HIGHEST Outstanding Satisfactory Poor
SCORE

1. General Appearance 3 Lines or patterns There are Smudges or stray

- Neatness and labelling are drawn clearly smudges or stray marks obscure details
and not smudged. pencil marks but of the illustration.
Minimal erasure or these do not Over-all quality of the
stray pencil marks. significantly affect drawing shows
Labelling is the over-all minimal effort to
accurate and appearance of the complete careful work.
complete. (3 illustration. Labels Important labels are
points) are included but missing. (1 point)
there are minor
problems in the
accurate
identification of a
part. (2 points)
2. Organization and 5 Provides complete Provides clear Only few important
content and well organized illustration contents are
-Illustrations and illustrations (5 however some illustrated (1 point)
sequencing points) details are missing
(3-4 points)
3. Attention to details 2 The illustration is Some minor errors Major discrepancies
accurately drawn are present are very noticeable in
(2 points) however these do the illustration. (no
not distract the point)
information
conveyed. (1
point)

CHAPTER I: Brief History of Forensic Chemistry & Toxicology

www.camdencc.edu www.forensicscienceeducation.org
Objectives
At the end of the topic, the students should be able to:
1.Identify the important dates involved in the advancement of Forensic Chemistry and Toxicology
2. Identify the important personalities involved in the advancement of Forensic Chemistry and Toxicology

1858 - 1st Medical Textbook Printed entitled “Manual de Medicina Domestica”

authored by Dr. Rafael Genard y Mas
1871 - Teaching of Legal Medicine started
March 31, 1876 - Creation of the position “Medico Titulares” virtue of Royal Decree No.
188 of the King of Spain
December 15, 884 - Creation of committee to study mineral waters of Luzon by Gen.
Joaquin Jovellar and Mr. Anacleto del Rosario as the chemist
1894 - Rules regulating the services of “Medico Titulares y Forences”
1898 - Preservation of the Spanish Forensic Medicine System by the
American Civil Government
1899 - Establishment of the first Crime Laboratory by the US Army
September 13, 1887 -Laboratorio Municipal de Manila was created
1894 -Creation of Laboratorio Medico Legal under the Judicial Branch of the
Government
1895 -Antonio Luna established a Clinical Laboratory that functions for
Chemical Analysis
1899 -First Scientific Laboratory on the Banks of Pasig River was established
headed by Lt. R.P. Strong
July 1, 1901 -Bereau of Government Laboratories was created
More in 1901 -Creation of Provincial Insular and Municipal Board of Health
1908 -Incorporation of the teaching of Legal Medicine and Ethics in the
Philippine Medical School (UP)

March 11, 1915 -Department of Legal Medicine was created

January 10, 1922 -The head of the Department of Legal Medicine and Ethics became the
Chief of the Medico- Legal Department of the Philippine General
Hospital without pay.
March 10, 1922 -Department of Legal Medicine of the UP as the branch of the
Department of Justice.
-Act No. 2465 of the Philippine
Legislature

October 14, 1924 -Legal Medicine as branch of the DOJ and at the same time an integral
part of UP

December 19, 1937 -Creation of the Division of Investigation under Department of Justice
March 31, 1938 -Department of Legal Medicine was Abolished and was turned over to
the
October 1939 -Philippine constabulary having its own medico-legal office with chemical
laboratory
1945 -Creation of the Criminal Investigation Laboratory with the office of the
Medical Examiner by the Provost Marshal of the US Army
1947 Ballistics, Photography and Fingerprint Record Unit was changed to
Criminal Laboratory Branch of the Constabulary
1951 Medico-legal Section was created under Col. Jesus T. Mendoza

CHAPTER II: Introduction to Forensic Chemistry

Objectives
At the end of the topic, the students should be able to:
1. Identify the different types and methods of evidence examination
2. Develop an understanding about the relevance of the subject in relation to crime investigation
3. Demonstrate belief on science in systematic process to help students in the easy identification of the
evidence.

Definition of Forensic Chemistry:

Application of Chemical Principles in the examination of physical evidences. Chemistry applied in
the elucidation of legal problems.

Roles of Forensic Chemistry:

a. Speedy Investigation
b. Solution of Crimes
c. Convicting the guilty
d. Clearing the Innocent

Functions of Forensic Chemistry:

a. Aid the investigator to determine if a specimen collected is effective for such examination
b. Analysis of: Blood, Metals, Poisons, Wines, Hair, Body Fluids, Textile, Gunpowder and Glass
Fracture
c. Able microscopists, physicist and photographer
d. Perception gathered by organ senses are imparted to others
e. Does not fall in any exemptions provided by the Rule of Court

Scope of Forensic Chemistry

a. Includes the chemical side of investigation
b. Analysis of material leading to legal proceedings
c. Not only purely chemical questions but aspects of Forensic Science

Stages in the Practice of Forensic Chemistry

1. Collection or Reception of the specimen to be examined
a. Sufficiency of the specimen
b. Standard for comparison
c. Maintenance for individuality
d. Labeling and Sealing
2. Actual Examination of Specimen
a. Scrutinize, document complete description of external appearance, manner of collection
and secured.
b. Take photographs if possible
c. Weigh, measure, record
3. Communication of Results
4. Court Appearance
a. Oral evidence is done if the case is brought to court

Factors Contributing to the loss of physical evidence

1. Lack of Precautions Preventing Tampering of Specimen
2. Failure in Preservation
3. Failure in transport of specimen
4. Failure in Identifying the specimen
5. Improper Packing of Specimen

The Six Golden Rules in Forensic Science

1. Go slowly 4. Consult others
2. Be thorough 5. Use imagination
3. Take notes 6. Avoid complicated theories
Types of Examination
1. Qualitative Examination which answers the question “What”?
2. Quantitative Examination which answers the question “How much”?

Techniques used in Forensic Chemistry

1. Microscopy - Magnify and resolve fine details
2. Photography - Preservation of evidence
3. Invisible Rays – Use of UV, IR and X-Ray
4. Chromatography - Separation of the constituents of a solution or colloidal dispersion
5. Electrophoresis - Migration of particles to opposite electrode
6. Spectrography – Elemental Analysis
7. Lase Technique – An innovation of spectrography
8. Mass Spectrometry – Analysis of sample in the Molecular weight
9. Spectrophotometry – measures concentration of various elements or compounds
10. Neutron Activation Analysis – Use of nuclear reactor
11. DNA Typing – DNA profiling
12. Forensic Entomology – Study of different insects
13. Atomic Absorption – Detect trace metals

Principles used in Forensic Chemistry

1. Law of Individuality - Every object, natural or manmade has an individuality which is not
duplicated in any other object
2. Law of Progressive Change - Everything changes with the passage of time.
3. Principle of Comparison - Only “likes” can be compared
4. Principle of Analysis - The analysis can be no better than the sample analyzed
5. Law of Probability - All identifications, definite or indefinite, are made consciously or
unconsciously on the basis of probability

Crime Scene Vocabulary

1. Primary Crime Scene - The original location of a crime or accident
2. Secondary Crime Scene - An alternate location, such as where additional evidence may be
found.
3. Suspect - Person thought to be capable of committing a crime.
4. Accomplice - Second person associated with committing a crime.
5. Alibi - Statement of where a suspect was at the time of a crime.
6. Evidence - Is a means, sanctioned by law, of ascertaining in a judicial proceeding the truth
respecting a matter of fact.
a. Testimonial Evidence - Would be any witnessed accounts of an incident or crime.
b. Physical Evidence - Any material items that would be present on the crime scene or the
victims. Presented in a crime investigation to prove or disprove the facts of the issue.
c. Trace Evidence - Refers evidence that is found at a crime scene in small but measurable
amounts.
d. Scientific Evidence - The means sanctioned by law, of ascertaining in a judicial
proceeding the truth respecting a matter of wherein scientific knowledge is necessary.
i. Real or Autoptic Evidence - Evidence which is addressed to the senses of the
court.
ii. Testimonial evidence - Comes from people. E.g. Testimony of an expert witness
in court
iii. Experimental Evidence - An expert witness may be required to perform certain
experiments to prove a certain matter of fact.
iv. Documentary Evidence - any written evidence presented by an expert in court.
e. Direct Evidence - Any fact to which a witness testifies based on what he saw, heard,
smelled, touched or tasted, is direct evidence.
f. Circumstantial Evidence - A kind of evidence which seeks to establish a conclusion by
inferences from proved facts
 g. Hearsay Evidence - A statement made by a witness on the authority of another and not
from his own personal knowledge or observation.
7. Witness - A witness in court may be an ordinary or expert witness
a. Ordinary Witness - He must have the organ and power to perceive the perception
gathered by his organ of sense can be imparted to others
b. Expert Witness - The opinion of a witness regarding a question of science, art or trade,
where he is skill therein, may be received in evidence.

Crime Scene Protocol

1. Interview - First step in processing a crime scene. To determine what allegedly happened, what
crime took place, and how was the crime committed.
2. Examine - Will help identify possible items of evidentiary nature, point of entry and point of exit,
and getting the general layout of the crime scene.
3. Photograph - Involves creating a pictorial record of the scene and record items of possible
evidence. Crime scene photographs taken in two categories (1) overall views and (2) items of
evidence.
4. Sketch - Demonstrate the layout of the crime scene or to identify the exact position of the
deceased victim or evidence within the crime scene
5. Process - Crime scene technician will process the crime scene for evidence, both physical and
testimonial evidence. Crime scene technicians to identify, evaluate and collect physical evidence
from the crime scene for further analysis by a crime laboratory.

Experiment No. 1
MATERIALS & GLASSWARES IN FORENSIC TOXICOLOGY

INTRODUCTION:
The forensic toxicology laboratory utilizes not only analytical instruments but pertinent glass wares,
materials and instruments. It is a must for the laboratory personnel to be acquainted and oriented with the
nature and uses of those that are commonly employed in the laboratory as these contribute in achieving
accurate, reliable and acceptable forensic results.

I. OBJECTIVES: At the end of the activity, the student will

- Identify the different laboratory instruments and materials essential in the conduct of Forensic
Chemistry Laboratory;
- Determine the different use/s of each material.

II. MATERIALS
a. Glass Slide and Cover Slip i. Cotton Swab
b. Lancet or Skin Pricker j. Staining Rack
c. Applicator Stick k. Gloves
d. Beaker (100 ml) l. Test Tube & Rack
e. Venipuncture Set m. Mixing Bowl
i. 10 ml Disposable Syringe n. Wire Gauze & Tripod
ii. 13 X 100 Test Tube o. Separatory Funnel
iii. Tourniquet p. Bunsen Burner
iv. Plaster q. Magnifying Glass
v. Tube holder r. Watch Glass
vi. Two-way needle s. Micropipette and tips
vii. Vacutainer Tubes t. Ultra-violet lamp
f. Pasteur Pipette u. Eye protector
g. Viewing Box v. Concavity slide
h. Serological Pipettes w. Surgical blade
- 1 ml Pipette x. Biohazard bag
- 5 ml Pipette y. Clinical Thermometer
- 10 ml Pipette z. Centrifuge & Ultracentrifuge

Rubric for Point Distribution:

Grading of laboratory experiments will utilize the 30-point scale. Grading will be based on the
following criteria:
1) Explanation and understanding of the science (5 points) - Does the report show sufficient
background information for the reader to follow and understand the experiments?
2) Figures and illustrations (5 points) - Are there sufficient photographs, tables and graphs necessary
to support the results. Are they properly labeled and referenced in the text?
3) Results (10 points) - Do the results of the experiment match the anticipated and expected and are
sources of variance properly noted and explained? Are all the points noted in the discussion
section covered?
4) Answers to questions (10 points) Grammar, organization, and references - Is the document clearly
written, well organized and properly referenced.

III. PROCEDURE

1. Examine the instruments/materials presented in the laboratory through a video recorded by the
instructor.
2. Determine the use/uses of these instruments and materials in Forensic Toxicology.
3. Draw neatly each material and instrument demonstrated by your laboratory instructor. Label all
parts accurately.

IV. ACTIVITIES
 - Draw and label the parts of the different glass wares, instrument and
Materials

V. QUESTIONS OF RESEARCH

1. Describe each and give the laboratory use or uses of each.

Experiment No. 2
INSTRUMENTATIONS IN FORENSIC TOXICOLOGY

INTRODUCTION:
The forensic toxicology laboratory utilizes analytical instrumentation applicable to training, routine and
specialized analytical and forensic case work. This instrumentation provides capabilities in robust and
reliable analysis of the varied physical evidences from the crime scenes and natural products, in addition
to the ability to conduct blood ethanol and semen analyses for forensic casework (Clark, 2008).

I. OBJECTIVES: At the end of the activity, the student will

- Identify the different laboratory instruments essential in the conduct of forensic toxicology
Laboratory tests;
- Determine the different use/s of the different instruments used in forensic toxicology.

II. MATERIALS: Illustrations, References on

1. Drunkometer/Intoximeter/Alcometer
2. Infrared spectroscopic
3. Fuel cell Devices
4. Spectroscopic Techniques:
a. Spectrometer
b. infrared Spectrogram
5. Chromatography
a. Gas Chromatography
b. GC/MS system

Rubric for Point Distribution:

Grading of laboratory experiments will utilize the 30-point scale. Grading will be based on the
following criteria:
5) Explanation and understanding of the science (5 points) - Does the report show sufficient
background information for the reader to follow and understand the experiments?
6) Figures and illustrations (5 points) - Are there sufficient photographs, tables and graphs necessary
to support the results. Are they properly labeled and referenced in the text?
7) Results (10 points) - Do the results of the experiment match the anticipated and expected and are
sources of variance properly noted and explained? Are all the points noted in the discussion
section covered?
8) Answers to questions (10 points) Grammar, organization, and references - Is the document clearly
written, well organized and properly referenced.

III. PROCEDURE

1. Illustrate and label the parts of the following instruments used in forensic toxicology.
2. Illustrate the operating principle of each instrument used in forensic toxicology.

IV. QUESTIONS OF RESEARCH

- Answer completely each question.

1. Describe and discuss the function of each instrument in forensic toxicology.
2. Illustrate and discuss completely the Mechanism of Function of each instrument in forensic
toxicology.
3. State the advantages and the disadvantages of automations in forensic toxicology laboratory.

Experiment No. 3
Safety and First-Aid in the Forensic Toxicology Laboratory

I. INTRODUCTION:
Safe laboratory practice according to Anderson (2015) ensures the well-being of laboratory personnel as
well as those who enter the laboratory for consultation and those responsible for cleaning the laboratory
and discarding hazardous waste. Laboratory safety includes a variety of policies. The system of
Standard Precautions of good working practices will reduce any danger of infection from any source:
- All blood samples and other human body fluids should be considered as a potentially infectious.
- Always use proper personal protective equipment to protect yourself when working with any body
tissue.

II. OBJECTIVES: At the end of the activity, the student will

- be familiar with the use of standard precautions in breaking the chain of infection;
- understand and be able to apply common laboratory safety rules;
- demonstrate the proper use of personal protective equipment;
- discuss and be able to apply proper first-aid techniques in response to poisoning emergencies.

III. MATERIALS: Reference books

Charts and illustrations

Rubric for Point Distribution:

Grading of laboratory experiments will utilize the 30-point scale. Grading will be based on the
following criteria:
1. Explanation and understanding of the science (5 points) - Does the report show sufficient
background information for the reader to follow and understand the experiments?
2. Figures and illustrations (5 points) - Are there sufficient photographs, tables and graphs necessary
to support the results. Are they properly labeled and referenced in the text?
3. Results (10 points) - Do the results of the experiment match the anticipated and expected and are
sources of variance properly noted and explained? Are all the points noted in the discussion section
covered?
4. Answers to questions (10 points) Grammar, organization, and references - Is the document clearly
written, well organized and properly referenced.

III. DISCUSSION (Saferstein, 2013)

A. PROTECTIVE CLOTHING:
Everyone who enters the laboratory should wear a laboratory coat. Replace laboratory coat immediately
if it becomes contaminated.
B. DISPOSABLE GLOVES:
Every sample handled in the laboratory is potentially hazardous and gloves should always be used when
handling toxic material including any body tissues. Replace gloves immediately if they are torn.
C. EYEWASHING:
Many infections can be easily caught by contact with the mucous membranes of the eyes. Wash your
eyes immediately with very large amounts of cold running water if contact with a possible infectious
material may have occurred
D. SHARPS:
Sharps, such as needles and lancets, should be disposed of in a special container that is spill proof,
puncture resistant and closable. The sharps container must be labeled in red or orange, must be
maintained upright and must display the biohazard symbol.

E. AEROSOLS:
Avoid all practices in the open laboratory which may cause splashing or the release of droplets or dust
into the air. Carry out all operation which cause aerosols in a suitable fume cupboard and wear safety
glasses.
F. TOXIC AND FLAMMABLE SUBSTANCES:
 Toxic and flammable materials must always be contained in a fume cupboard or a suitable safe box.
G. ELECTRICAL EQUIPMENT:
Take special care with any equipment which uses liquids. Always leave the installation, servicing and
repairs to qualified personnel.
H. PERSONAL:
Avoid touching your face or mucosae (eyes, nose, and mouth) with your hands while in the laboratory.
You must not eat, drink, smoke or put-on make-up while in the laboratory. Always wash hands
thoroughly before leaving the laboratory.
I. ACCIDENTS:
All spills should be cleaned immediately with appropriate disinfectant solutions (e.g. dilute bleach
solution), and materials used to wipe up spills should be disposed of as potentially biohazardous. Each
laboratory has a cleanup procedure, but general recommendations include:
 Wear gloves
 Use dilute (1:10) bleach solution or commercially prepared solution.
 First clean area with visible blood then disinfects the entire area of possible
contamination.
 Keep the bleach in contact with the contaminated area for at least 20 minutes to
ensure complete disinfection.
 All accidents should be reported immediately to the instructor/supervisor in-
charge.

IDENTIFICATION OF HAZARDS:
Hazard is the potential of a substance to cause harm.
The risk from that substance is the likelihood of it harming someone under the conditions of use:
A. BIOLOGIC HAZARDS
Blood, urine, spinal fluid and all other body fluids present biologic safety hazards because they may
contain highly infectious and potentially lethal organisms or viruses.
B. CHEMICAL HAZARDS
Solid, liquid or gaseous chemicals may be hazardous if transported, handled, stored or dispensed
inappropriately. Chemicals may have toxic, flammable, or carcinogenic properties.
C. ELECTRICAL HAZARDS
Electrical hazards are caused by inappropriate use or maintenance of electrical instruments or equipment
that can cause electrical shock, burns or a fire or explosion.
D. MECHANICAL HAZARDS
Mechanical hazards may result from improper use, storage or disposal of glassware, sharp instruments,
compressed gases or equipment.
E. FIRE HAZARDS
Fire or thermal hazards may result from the improper use, storage of either cryogenic substances or
substances capable of combustion. Fires can obviously cause burns, and skin contact with cryogenic
substances has essentially the same effect – it causes a thermal burn. Cryogenic and combustible
substances may cause a fire, explosion, or asphyxiation.

IV. PROCEDURE

Draw/illustrate the following:

1. Biohazard Sign
2. Safety Signage
 3. Personal Protective Equipment
4. NFPA Rating System

V. QUESTIONS OF RESEARCH

1. Enumerate, draw and explain briefly the meaning of signages which can
lessen injury in the laboratory.
2. Describe the first-aid treatments in poisoning cases.
3. Discuss the methods of decontamination in cases of ingested poisoning.

Assessment No. 1

Part 1: Define the following

1. Medico Titulares
2. Medico Legal

3. Philippine Constabulary

4. Provost Marshall

5. Bureau of Government
Laboratories

Part 2: Arrange the Crime scene Protocol, from 1 being the first stage to 5 being the last stage.

Demonstrate the layout of the crime scene or to identify the exact position of the
deceased victim or evidence within the crime scene
To determine what allegedly happened, what crime took place, and how was the
crime committed.
Crime scene technician will process the crime scene for evidence, both physical
and testimonial evidence.
Involves creating a pictorial record of the scene and record items of possible
evidence.
Will help identify possible items of evidentiary nature, point of entry and point of
exit, and getting the general layout of the crime scene.

Part 3: Identification

Study of different insects

Any fact to which a witness testifies based on what he saw, heard, smelled,
touched or tasted, is direct evidence.
All identifications, definite or indefinite, are made consciously or
unconsciously on the basis of probability
A type of witness, the opinion of a witness regarding a question of science,
art or trade, where he is skill therein, may be received in evidence
A statement made by a witness on the authority of another and not from his
own personal knowledge or observation
A technique used in forensics that preservation of evidence
A technique used in forensics which measures concentration of various
elements or compounds.
Person thought to be capable of committing a crime.
Statement of where a suspect was at the time of a crime.
A principle in Forensics which states that every object, natural or manmade
has an individuality which is not duplicated in any other object

CHAPTER III: Blood and Blood Stains

Objectives
At the end of the topic, the students should be able to:
1. internalize the importance of blood and blood stain in crime investigation.
 2. identify the different blood groups present in blood
3. compare the stages of blood and blood stain examinations

Importance:
• As circumstantial or corroborative evidence against or in favor of the perpetrator
• For disputed parentage
• Determination of the cause of death and the length of time the victim survived the attack
• Determination of the direction of escape of the victim or the assailant.
• Determination of the origin of the flow of blood.
• Determination of the approximate time the crime was committed

Nature of Blood:
a. Largest circulating tissue of the body
b. Consists of vital substances
c. Fluid that circulates into the Cardiovascular System (CVS)

Kinds of Blood:
a. Arterial Blood b. Venous Blood
- Aka Capillary blood - Dark Red in color
- Bright red color - Contains increased amount of CO2
- Oxygenated blood - Non-oxygenated blood

Characteristics of blood
a. Color – Bright red for arterial blood and dark red for venous blood
b. Volume – 70% of Total body weight
c. Viscosity – Resistance to blood flow
- Blood is thick and sticky
- Normally flows with difficulty
d. Specific Gravity – Weight of blood compared to water on the same volume
- Distilled Water: 1.000
- Blood: 1.065 (due to cellular elements)
e. pH Reaction – Slightly alkaline (7.35-7.45)
f. The circulating tissue of the body - 1 cc of blood: 5,000,000 red cells
- Man of average size: about 6 quarts of blood

Composition of Blood

Formed Elements:
- Composing about 35% of the total blood volume
a. RBC (Erythrocytes)
- Number: 5,000,000 RC/mm3
- Diameter: 7-10 microns
- Rate of Destruction:
- 10 Billion Cells/hr in Adults
b. White Blood Cells/Leukocytes
- Number: 5-10,000/mm3
- Soldiers of the body
- Resists attacks of diseases
c. Platelets or Thrombocytes
- Number: 150-350,000/mm3
- Functions for blood coagulation
Liquid Portion:
- 65% of the total blood volume
a. Plasma – Straw liquid portion of unclotted blood
b. Serum – Straw-yellowish liquid that separates when blood is allowed to clot

Hemoglobin:
- Coloring matter of blood
- Pigment which is found at the cytoplasm of Red Blood Cells

Types of Hemoglobin
a. Abnormal Derivatives of Hemoglobin
a. Methemoglobin (HbM) – Found in NO3 and NO2 poisoning with a chocolate brown color
b. Sulfhemoglobin (HbS) – Found in the presence of bacteria, severe constipation,
enterogenous cyanosis and the blood color is lavender
c. Carboxyhemoglobin (HbCO) – Due an excessive inhalation of gas from defective stoves
and from automobiles and imparts cherry red color of blood
b. Normal Hemoglobin
a. Oxyhemoglobin (HbO2) – Hemoglobin combined with oxygen and which gives the color
to the arterial blood
b. Reduced hemoglobin -Hemoglobin combined with carbon dioxide and which gives color
to venous blood

Method of Collecting Blood

A. Capillary Blood Sample
a. Collected in the area of the skin/finger/ring puncture
b. Collection of arterial blood
c. Small quantity of blood with the use lancet or pricker
i. Puncture sites:
1. Ring finger (Adult and children)
2. Ear lobe (Adults)
3. Heal or Toe (Infants and Children)
B. Venous Blood Sample
a. Collection of larger volume of blood and can be taken from several veins
i. Cephalic vein
ii. Medial cephalic vein
iii. Basilic Vein
iv. Jugular Vein
C. Arterial Blood Collection
a. Collection of large volumes of blood collected from the arteries
i. Radial Artery
ii. Brachial Artery
iii. Femoral Artery

Tests to Identify Blood Stains

Questions to be answered:
a. Does the stain contain blood or other substances?
b. Is the stain being that of blood
 c. If the stain is of human blood, did it come from the victim, the accused or from other persons?
d. Is it human or animal?

Screening Tests for Blood and Blood Stain

a. Benzidine Test b. Guaiacum Test c. Phenolphthalein d. Precipitin e. Blood Typing

Test Test
-Very delicate test - Van Deen’s or - Aka Kastle Mayer -Test for protein -Identify specific
-Detects blood when Schoenbein’s Test Test found in human blood type of a
present in a -Reagents: Guaiac -Reagent: 2g blood sample
dilution of 1:300,000 Reagent and 3% Phenolphthalein, 10- -Antibody or -May serve many
parts H2O2 30g Zinc and 20g precipitin reacts purposes
-Positive result: Blue -Positive Result: KOH with human
Color Rapidly Blue Color -Positive result: blood forming
Develops Permanganate color Precipitates
-Positive result:
Gray
precipitation ring
after 20 mins

Blood Grouping
1. ABO Blood Grouping – First blood group system and the only blood Group system that can be
determined using serum
- Blood type depends on:
- Surface protein Antigen
- Antibodies to these substances
- 4 Blood Types:

Methods of Blood Typing

1. Direct or Forward Typing
- Antigens A and B will always react with its specific antibody in the serum causing agglutination
- Makes use of A and B Antisera that came from lectins and capable of agglutinating A and B
antigens
 2. Indirect or Reverse Blood Typing
- Blood type of the serum is the opposite of the antigen where agglutination occurs

Confirmatory tests for Blood and Blood Stain

a. Teichmann or Haemin Crystal Test b. Takayama Test
- Makes use of NaCl and 2-3gtts GHAc - Aka Haemochromogen test
- Positive result: Dark Brown Rhombic Crystals = Positive result: Salmon pink colored Rhombic
arranged singly or in cluster crystals arranged in cluster or sheaves.

Death due to loss of blood

- Death due to loss of blood can be due to different causes like gun shot wound or severe blunt trauma
- This will lead to decreased blood pressure and decreased rate of bleeding
- Blood and blood stains can be analyzed from the scene of the crime, from the clothing and from the
body itself
- For person who lived for a considerable amount of time, large pool of blood from small wounds can be
found and usually the cause of death for this is hemorrhage

Blood Spatter Analysis

1. Direction of Escape - Blood striking a smooth surface leaves a large blot and with one or two
smaller blots trailing off in a straight line.

2. Blood from the Neck – Swagger away and large forms of blots on the ground
 3. Cuts from small arteries – Spurt of blood in a defined pattern

4. Drops of Blood striking obliquely – Head is on the direction of the flight of drop

Person is Bleeding Profusely and struggle has taken place

Experiment No. 4
SPECIMEN COLLECTION AND HANDLING
(Venipuncture)

INTRODUCTION
In forensic toxicology, blood samples that are taken from convicted suspects are determined. The blood
gets testified and diagnosis for any sign of evidence that were affiliated with the crime scene's aftermath.

Phlebotomy is the practice of using a needle to withdraw a sample of blood from a designated vein. In
earlier times, phlebotomy was called bloodletting. It was used to treat and/or cure diseases. Phlebotomy
today is also called venipuncture or venipuncture. A phlebotomist is a person who performs phlebotomy.

Another field of phlebotomy is forensics, where criminal pathologists examine the blood of crime victims to
determine the exact cause of injury and/or death. Phlebotomists also are given the task of insuring that all
blood samples are guarded safely against contamination and/or tampering (http://www.ehow.com/
about_5505415_definition-phlebotomy.html).

I. OBJECTIVE: At the end of the activity, the student will

- know the different methods of blood specimen collection as applied in forensics;

- perform accurately and precisely the phlebotomy techniques appropriate for forensic
professionals.

II. MATERIALS
1. Phlebotomy set
a. Syringe Technique b. Vacutainer Technique
- disposable syringe (10 ml) - tube holder
- tourniquet - 2-way needle
- glass tubes (13 X 100) - vacutainer tubes
- applicator stick a. red top
- wet and dry cotton balls b. lavender top
- plaster - Gloves
2. Reagent: - Masks
- betadine
- 70% Ethyl alcohol
3. Instrument
- Clinical centrifuge
- Forcep (curve)

Rubric For Point Distribution:

Grading of laboratory experiments will utilize the 30-point scale. Grading will be based on the
following criteria:
1.Explanation and understanding of the science (5 points) - Does the report show sufficient
background information for the reader to follow and understand the experiments?
2. Figures and illustrations (5 points) - Are there sufficient photographs, tables and graphs
necessary to support the results. Are they properly labeled and referenced in the text?
3. Results (10 points) - Do the results of the experiment match the anticipated and expected and
are sources of variance properly noted and explained? Are all the points noted in the discussion
section covered?
4. Answers to questions (10 points) Grammar, organization, and references - Is the document
clearly written, well organized and properly referenced.

III. PROCEDURE

1. The instructor orients the students with the different materials, glass wares and
instruments used in phlebotomy through a live video stream or a recorded video;
2. The instructor demonstrates the procedures in performing phlebotomy techniques;
3. The instructor presents the procedures of specimen handling and preparation.
A. PERFORMANCE OF A VENIPUNCTURE: (Calaluce,2015).

 Palpate and trace the path of veins with the index finger. Arteries pulsate, are most
elastic, and have a thick wall. Thrombosed veins lack resilience, feel cord-like, and roll
easily.
 If superficial veins are not readily apparent, you can force blood into the vein by
massaging the arm from wrist to elbow, tap the site with index and second finger, apply a
warm, damp washcloth to the site for 5 minutes, or lower the extremity over the bedside
to allow the veins to fill.
 Approach the patient in a friendly, calm manner. Provide for their comfort as much as
possible, and gain the patient's cooperation.
 Identify the patient correctly.
 Properly fill out appropriate requisition forms, indicating the test(s) ordered.
 Verify the patient's condition. Fasting, dietary restrictions, medications, timing, and
medical treatment are all of concern and should be noted on the lab requisition.
 Check for any allergies to antiseptics, adhesives, or latex by observing for armbands
and/or by asking the patient.
 Position the patient. The patient should either sit in a chair, lie down or sit up in bed.
Hyperextend the patient's arm.
 Apply the tourniquet 3-4 inches above the selected puncture site. Do not place too tightly
or leave on more than 2 minutes (and no more than a minute to avoid increasing risk for
hemoconcentration). Wait 2 minutes before reapplying the tourniquet.
 The patient should make a fist without pumping the hand.
 Select the venipuncture site.
 Prepare the patient's arm using an alcohol prep. Cleanse in a circular fashion, beginning
at the site and working outward. Allow to air dry.
 Grasp the patient's arm firmly using your thumb to draw the skin taut and anchor the vein.
The needle should form a 15-to-30-degree angle with the surface of the arm. Swiftly
insert the needle through the skin and into the lumen of the vein. Avoid trauma and
excessive probing.
 When the last tube to be drawn is filling, remove the tourniquet.
 Remove the needle from the patient's arm using a swift backward motion.
 Press down on the gauze once the needle is out of the arm, applying adequate pressure
to avoid formation of a hematoma.
 Dispose of contaminated materials/supplies in designated containers.
 Mix and label all appropriate tubes at the patient bedside.
 Deliver specimens promptly to the laboratory.

IV. ACTIVITIES
- Schematically illustrate the preparations of serum and plasma.
- Label according the layers of blood formed using the following methods of blood extraction:
a. syringe method;
b. vacutainer method

V. QUESTIONS FOR RESEARCH

1. Discuss the difference between macro and micro blood collection.
2. Differentiate completely the difference of an Open and Closed system of blood collection.
3. What are the complications of venipuncture? Give the remedy of each.
4. What are the sources of biases in venipuncture?
5. Discuss how the following samples are collected for forensic determinations:
a. Urine for males and females
b, Cerebro Spinal Fluid
c. Sweat
d. Semen
 e. Saliva
6. Differentiate syringe method of collective blood from vacutainer method.
 Experiment No. 5
Preliminary Test for Blood and Blood Stains
Direct ABO Blood Grouping

Introduction
The most well-known and medically important blood types are in the ABO group. They were
discovered in 1900 to 1001 at the University of Vienna by Karl Landsteiner in the process of trying to
learn why blood transfusion sometimes cause death and at other times save patients.
There are four principal blood types: A, B, AB and O. There are two antibodies that are mostly
responsible for the ABO blood types. The specific combination of these four components determines the
blood type of an individual. An ABO blood typing is a procedure performed to determine someone’s blood
type. This is performed using a direct or a reverse method. For this activity, a direct ABO blood grouping
will be performed.

Objectives: At the end of the activity, the students are expected to be able to:
a. appreciates how to perform accurately the procedure for the direct ABO blood typing and Rh
blood typing
b. understands the principle behind ABO and Rh blood grouping

Materials:
a. Lancet g. Anti-A reagent
b. Wet and Dry cotton h. Anti-B reagent
c. Glass slide
d. Applicator stick
e. Viewing box
d. Biohazard bag and gloves
f. Pasteur pipette

Procedure: The instructor will be recording a video of the procedure and to be uploaded in Canvas.
Direct ABO Blood Grouping:
1. Materials and reagents will be assembled.
2. Sterilize the puncture site of the ring finger using 70% isopropyl alcohol by running against the skin to
remove microorganism.
3. Air dry the area sterilized. Do not touch with any unsterile surface.
4. Apply a light pressure at the site with the thumb making the skin firm and puncture with pressure
applied.
5. Release the pressure applied.
6. Wipe the first drop of blood with a dry sterile cotton.
7. Apply a slight pressure to form a drop of blood.
8. Dub 0blood sample with the initially prepared slide by placing one drop each in compartments A and B.
9. Place the slide on a flat surface.
10. Dispense one drop of Anti-serum A and Anti-serum B in each compartment respectively.
11. Mix the blood and anti-serum with applicator stick. A separate applicator stick is used to mix the blood
in two preparations.
12. Swirl the entire slide for at least two minutes or until agglutination has formed.
13. Observe for agglutination or clumping of cells in the view box and in the microscope.

Activity:
1. Draw the result of the forward slide method for blood type A, B, AB and O based on their agglutination
reaction.

Questions for research:

1. What are the lectins used in the A-antiserum and B-antiserum?
2. What are the possible offspring of ABO blood type?
 Experiment No. 6
Confirmatory Test for Blood and Blood Stain
Microscopic Examination

Objectives: At the end of the activity, the students are expected to be able to:
a. Identify the sources and natures of an alleged blood and blood stains and distinguish
between human, avian and piscine blood samples.
Materials:
A. Specimen: Prepared blood slides of the following
a. Human Blood (fresh and old)
b. Avian Blood (Chicken’s blood)
c. Piscine Blood (Fish’s blood)
d. Menstrual Blood
e. Wright’s stain

B. Materials:
a. Electric Microscope

Procedure: The instructor will be recording how to focus the different slide. The image of the different
blood smears under the microscope will shown in the video. The video recording will be uploaded in
Canvas. Student may use additional online and book references to help them in this activity.

Activity:

Drawing/Illustrations

1. Draw/illustrate the different microscopic appearance of all the slides prepared.

a. Human Blood (fresh and old)
b. Avian Blood (Chicken’s blood)
c. Piscine Blood (Fish’s blood)
d. Menstrual Blood

Questions for research

1. Describe completely the different blood in terms of their shape and size.
2. What are the common stains used for blood smear?
 Experiment No. 7
Confirmatory Test for Blood and Blood Stain
Micro Chemical Examination

Introduction

Many different tests have been used to confirm that a stain contains blood. The oldest is chemical
confirmation of the presence of hemoglobin or its derivatives by the formation of specific crystals. For
example, the Takayama or hemochromogen test, in which ferrous iron from hemoglobin reacts with
pyridine to produce red feathery crystals of pyridine ferroprotoporphyrin. Another confirmatory test uses
the Teichman reagent, consisting of a solution of potassium bromide, potassium chloride and potassium
iodide in glacial acetic acid, and is heated to react with hemoglobin. The reaction first converts the
hemoglobin to hemin, and then halides react with the hemin to form characteristic brownish-yellow
rhomboid crystals.

Blood can be identified as being of human origin by precipitin reactions with antisera specific for
components of human blood. Usually this is an anti-human serum. Strictly speaking, this is a test for
human origin not for human blood, as serum constituents such as albumin and some globulins are found
in the extra vascular space.

Objective: At the end of the activity, the students are expected to be able to:
1. appreciate how to properly perform the micro-chemical methods for blood and blood stain
determination.
2. to understand the principle of each tests.

Materials:
a. Blood stain e. Pyridine reagent
b. Watch glass f. NSS
c. Glass slide with cover slip g. Saturated glucose solution
d. Electric microscope h. Glacial acetic acid

Procedures: The instructor will be recording a video demonstrating how to properly perform the different
micro-chemical test. The video will be uploaded in Canvas.

A. Preparation of Takayama Reagent

1. In a 50-mL beaker, dispense 3cc. of 10% NaOH, 3 cc. of pyridine and 3 cc of saturated
glucose solution
2. Add 7 cc of distilled water.

B. Procedure for Takayama Test

1. Cut a piece of known blood stain and place it in a watch glass.
2. Add 5mL of normal saline solution and tease with pins.
3. Add 2 mL more of normal saline solution and allow to stand for 5 mins.
4. Separate the fibers as a thin layer and add 1 mL of the Takayama reagent to the remaining
fluid
5. Aspirate sample fluid on a glass slide, cover with cover slip and view under the microscope.
6. Note the formation of crystals. Large rhombic crystals of salmon pink color, arranged in
clusters or sheaves appear with in 6 mins. Heat maybe applied to hasten crystal formation.
C. Teichman test
1. A small crystal of NaCl and 10 drops of glacial acetic acid are placed on a slide with a minute
amount of blood stain
2. Heat the preparation to crystallize the blood
3. Focus under the microscope using the HPO
4. Note dark rhombic crystals of haemin chloride are seen in singly or in cluster.

Activity:

Drawing/Illustration
1. Draw/illustrate Crystals form in the Takayama test
2. Draw/illustrate the crystals formed in the Teichamann test

Questions for Research

1. Explain the principle of Takayama test
2. Explain the principle of Teichman test
 Assessment No. 2

Part 1: Identify the Blood type using the forward typing

Blood type Anti-A Anti-B

With agglutination With Agglutination

Without Agglutination With Agglutination

With Agglutination Without Agglutination

Without Agglutination Without Agglutination

Part 2: Identify the Blood type using the reverse typing

Blood type A antigen B antigen

With Agglutination Without Agglutination

With Agglutination With Agglutination

Without Agglutination With Agglutination

Without Agglutination Without Agglutination

Part 3: Match the positive results of each tests used to screen the presence of blood and blood
stain

A. Guaiacum test B. Benzidine C. Pricipitin Test D. Blood typing E. Phenolphthalein

test

Also known as Van Deen’s test

Identify specific blood types
This test for proteins present in human blood
Also known as Kastel Mayer test
Positive result is blue color that rapidly develops.

Part 4: Differentiate Teichman’s from Takayama test

 CHAPTER IV: Seminal Fluid Analysis

Biological Characteristics of Semen

Typical Ejaculate
 2-5 mL of semen with approximately 160 million sperm
o This may contain 3pg DNA/Sperm = 480,000 ng DNA/Ejacualte
o Only 1 ng DNA needed for STR typing
 Seminal Fluid
o Medium for ejaculation
 Methods of collection: Self production, Condom collection, Aspiration from
vaginal vault after coitus and Coitus interruptus
 Samples must be collected in a wide-mouthed bottle
o May contain enzymes and other proteins
 Enzymes includes: Acid phosphatase, Prostate specific antigen and semenogelin
o Contains Sperm cells or spermatozoa
o Odor: Musty, Acrid and Fishy
o Color: Pearly white/ Gray-white and slightly turbid
 Increased white turbidity indicates infection and WBCs
 Yellow color indicates contamination of urine, prolonged abstinence and some
medications
o Viscosity: Highly Viscid
o Liquefaction is usually within 30 to 60 minutes
 Viscosity reporting is as follows
 0 (watery) to 4 (gel like)
 Increased viscosity and incomplete liquefaction: Impede sperm motility
 Prolonged liquefaction will result to decreased prostatic enzymes
o pH is 7.2 to 8.3
 Increase in pH may mean infection in the reproductive tract and decreased pH
may mean increased prostatic fluid
o Specific Gravity is between 1.027 to 1.032
 Semen is an extremely good source of DNA, but…
o Not all semen stains may contain sperm
 Vasectomy is a procedure undergone by males where sperm flow is blocked
from being ejaculated, and because of this, DNA typing is not possible.
 Cases of infertility where the severity is a factor. DNA typing for this may be
possible.

The Spermatozoa
 Has three distinct regions:
o Head – this contains the acrosome and nucleus (contains haploid DNA)
o Middle Piece - this contains the Mitochondria
o Tail – Also known as the flagella responsible for its movement
Forensic investigation for Seminal Fluid

1. Overlay Method
a. Spray a Whatman filter paper with distilled water.
b. Lay the paper down over the suspected semen stain and leave it for about 30 to 60
seconds
c. Remove the filter paper from the stain and observe for color change to Purple
2. Spot Test Method
a. Wet a sterile cotton swab with distilled water then roll swab against the stain
b. Saturate the swab with Acid Phosphatase solution

3. MUP (Methylumbelliferone phosphate method)

a. A more sensitive test than acid phosphatase
b. AP catalyzes the removal of the phosphate residue on the surface of the substrate 4-
methylumbelliferone phosphate which generate fluorescence under UV light.
c. Followed by a filter paper overlay, but instead of distilled water, paper is sprayed with MUP
d. Stain will be subject to UV lamp and Fluorescence of stain will be detected.

4. Test for Prostate Specific Antigen (PSA)

a. This is the major protein found in seminal fluid
b. Detected using immunochromatography test strip assay
5. Test for Semenogelins
a. Higher concentration in seminal fluid than PSA and not found in any other body fluids
b. Has a greater specificity for semen than PSA
c. Detected with the use of immunochromatographic test strip
6. Microscopic Examination (Part of Laboratory Discussion and QFR)
7. Micro-Chemical Examination (Part of Laboratory Discussion and QFR)
a. Florence Test
b. Barberio’s Test
 Experiment No. 8
Seminal Fluid Analysis
Physical and Microscopic Examination

Introduction

Determining the presence or absence, identity and distribution of particular body fluids is key to
the investigation of crimes against the person e.g. rape, assault or murder. The body fluids encountered
in these types of cases are usually blood, semen and saliva. The correct identification of traces of these
fluids at the crime scene or clothing maybe vital to a criminal investigation and the presence and absence
of a particular fluid may assist forensic biology expert in determining why the crime has occurred and
identify the persons who are involved and not involved with the crime.

Microscopic examination of sperm is more confirmatory method of discovery. Using microscopic

equipment, it is often possible to view the sperm cells, proving their presence.

Objectives: At the end of this activity, the students are expected to be able to:
a. describe the physical and microscopic characteristics of the semen
b. realize the importance of semen analysis in the investigation of crime against the person
c. determine the morphological characteristics of sperm cells
d. appreciate

Materials:
a. Freshly ejacuated semen k. Cover slip
b. Test tubes l. Red and blue litmus paper
c. Serologic pipette m. Gloves
d. Electric microscope n. Biohazard container
e. Glass slide m. 95%ethanol
f. Beaker o. Wrights stain
g. Pasteur pipette p. Distilled water
h. Watch glass q. Hematoxylin and eosine
i. Tally counter r. Petroleum gelly
j. Pipetol

Procedure

A. Physical Examination
The instructor will be discussing the physical characteristics of the seminal fluid. The students
must take down these important characteristics.
a. Color: ___________________________________________
b. Odor: ___________________________________________
c. Viscosity and liquefaction: ___________________________
d. Volume: _________________________________________
e. pH: _____________________________________________

B. Microscopic Examination
The instructor will be discussing the microscopic characteristics of the sperm. The students must
take note of these important characteristics.
 a. Sperm morphology: a smear of seminal fluid will be prepared by spreading a small
amount at the center of the slide.
b. Slides are air dried completely and fix the smear by immersing the slide several times in
methyl alcohol.
c. Hematoxylin and eosin staining will then follow.
d. Slides will then be focused under oil immersion objectives.
e. Morphological characteristics of will then be observed
 Head’s width and length: _____________________________________
 Midpiece width and length: ___________________________________
 Tail width and length: _______________________________________

Activity:

Drawing/Illustration:
1. Draw/illustrate the microscopic appearance of a sperm. Label the parts of the sperm.

Questions for Research:

1. What is the importance of semen analysis in sexual crime investigation?
2. Define the following terms:
a. Azoospermia
b. Oligospermia
c. Necrospermia
3. What is a Retrograde ejaculation?
4. Enumerate and discuss the three important criteria in semen ejaculation.
5. What is Florence test and discuss its principle?
6. What is Barberio’s test and discuss its principle?
 Assessment No. 3

1. Differentiate Azospermia from Necrospermia.

2. What is Sperm motility?

3. What is Oligospermia?

4. Enumerate the different types of abnormal sperm

5. How important is seminal fluid analysis in Forensic investigation? Explain your answer.
 CHAPTER V: Forensic Analysis of Hair

History of Hair Examination

 This is one of the oldest forms of Physical Evidence
 First used as physical Evidence in the year 1847

1891 - Han Gross 1897 - Rudolph Virchow became 1906 - Hugo Marx
published the first description of the the first person to do an in-depth wrote a paper on the use of hair in
uses of physical evidence to help study of hair. forensic investigations to determine
solve crimes identity.

1916 - Albert 1920 - Locard becomes known for 1931 - Dr. Paul Kirk works on new
Schneider became the first to the exchange principle – the fact ways to improve the use of hair in
collect physical evidence with a that “every contact leaves a trace.” forensic investigation
vacuum.

Hair Evidence
 Composed primarily of the protein keratin
 Each species of animal possesses hair with characteristic length, color, shape, root appearance,
and internal microscopic features that distinguish one animal from another
 Considerable variability also exists in the types of hairs that are found on the body of an animal.
 Hairs found on the head, pubic region, arms, legs, and other body areas have characteristics that
can determine their origin.
 Hairs can be transferred during physical contact
 The difference between black and brown hair is the amount of melanin
 Physical contact may result in the transfer of hairs.

Hair in Forensic Chemistry

 Identify both dead and the living
 Identify human from animal hair
 Identify the perpetration of a crime
 Comparison of the microscopic characteristics of questioned hairs to known hair samples helps
determine whether a transfer may have occurred.
o Assists in Rape, murder, assault, kidnapping etc.

Hair anatomy and Growth – present on many different regions of the body
a. Head d. Axilla
b. Pubic area e. Limbs
c. Chest

Hair is not present in the following area:

a. Palms b. Hands c. Soles of feet
Cyclic growth of Hair
A. Anagen Phase (80-90%) B. Telogen Phase (10-18%) C. Catagen Phase (2%)
Actively growing Follicle is dormant or resting Transition period between
Materials deposited in Hairs are routinely lost the anagen and telogen
hair shaft primary source of evidentiary phases
material.
Major Components of Hair

A. Hair Roots
 Portion embedded on the skin
 Can be either dry or dead or living roots (seen in hair in full growth)

Naturally shed hairs, such as a A hair forcibly removed from the Forcibly removed hairs may
head hair dislodged through scalp will exhibit stretching and have tissue attached.
combing, display undamaged, damage to the root area.
club-shaped roots.

B. Shaft – Present above the surface of the root and the most distinct part
1. Cuticle – outermost covering and a layer of non-nucleated polygonal cells. This usually
appears as overlapped (Scales of fish appearance)
 Most useful in the investigation of the origin of the hair sample
 Several patterns maybe observed

A. Coronal or Crown like Spinous or petal like Imbricate or flattened

2. Cortex – This is the thickest layer of the hair shaft which can be either straight or crosswise
 Examined for the presence of coloring matter in the form of tiny granules
 Pigment bodies may contain:

Cortical fusi - Air spaces of varying sizes found Granules- Small, dark, granulated structures that
near the root of a mature human hair vary in size, color, and distribution. Typically
distributed toward the cuticle in humans.
 3. Medulla – The central canal that contains the pigmented cells and absent in fuzz hair.
 The medulla may be: Continuous, Fragmented or interrupted

 In human hairs, the medulla is generally amorphous in appearance or completely absent.

 In animal hairs, its structure is frequently very regular and well defined.

human hair with no medulla. hair with trace medulla. Photomicrograph of a hair
with a clear, continuous
medulla

C. Tip or point – Appears blunt, round or frayed. This shows whether the hair has been cut.
 Hair tips for women usually has fine tips while m en has a cut off square

Collection of Hair Specimen

 Search for and collection of hair evidence should begin as soon as possible.
o Hair evidence is easily transferred to and from the crime scene.
 Collection should be done by:
o Hand if the location of the hair is important
o Sticky tape and Lint rollers
o Special filtered Vacuum Cleaner
  collect hairs and fibers end mass from carpet, bedding, etc.
 Hair evidence should be packaged into paper packets
 If sticky tape or a lint roller are used, the entire surface used should be packed into a
polyethylene storage bag
 Control samples need to be collected from the victim, suspect, and other individuals.
 Take from all pertinent regions of the body:
o 50 head hairs
o 24 pubic hairs
 Root still intact is preferable.
 Hair evidence should be looked for in the following: Clothing, Combs, Weapons, Pockets, Fingers
and Hat
 Get samples from both victim and suspect
o For dead body: hair must be collected from the head and the pubic area
 Best way of collecting hair is by Combing

Preservation of Hair
 Methods of packing hair
o Pill Box or Test tube – for questioned specimen
o Druggist powder paper – Properly folded, sealed and labeled

Examination of Hair
 Check for the following
Color Kind of cuticle
Length Cortex
Character (Wiry, Wavy, kinky) Medulla
Thickness Presence of dyes and bleaches
Kind of tip Cross section
How did it fall? Medullary index
Condition of root
 Difference between Human hair and Animal Hair

Human Hair Animal Hair

Medullary Index less than 0.5 Medullary Index more than 0.5
Medulla may not be present Medulla always present
Scale pattern is fine, overlaps more than 4/5 Scale pattern is coarse, overlaps less than 1/2

Pigment granules are Pigment granules are

fine course
54

Microscopic examination of Hair samples

Comparison Microscope Scanning Electron Microscope
Link the suspect to a crime scene. Determine the species, race, and somatic origin of
Control Hair match that of the suspect. a hair.
Exclude the suspect from a crime scene, meaning In addition:
that a control hair does not match the evidential A. DNA on the follicular tag
hair. B. Drug test
C. Environmental toxins

Used in Forensic Toxicology:

 to test and determine whether a drug was used.
 A drug that is ingested, enters the blood stream and is broken down to a specific metabolite.
 Hair strands normally grow at an average rate of 1.3 centimeters every month; they absorb
metabolized drugs that are fed to the hair follicle through the blood stream.
 Drug will only disappear if exposure to the drug is ceased, and the hair containing the drug is cut.
 Two Assays used in Forensics: Radioimmunoassay (RIA) and Enzyme-linked immunosorbent
assay (ELISA)

DNA Analysis of Hair sample - Can be extracted from the root or follicular tag of an anagenic hair
 Nuclear DNA (nDNA) – Comes from both patients and lead to individualization
 Mitochondrial DNA (mtDNA) – passed only from mother to offspring

Body Area Determination

A. Head hair longest hairs on the human body

uniform diameter and, often, a cut tip
Appear uncut, with tapered tips
B. Pubic hair Coarse and wiry in appearance
Exhibit considerable diameter variation or buckling
Often have a continuous to discontinuous medulla
Tapered tips are common, these hairs may also be abraded or
C. Facial hair Called beard hairs or mustache hairs.
Coarse in appearance
Triangular cross section
Wide medulla
Razor-cut tip.
D. Limb hair Legs and arms
Shorter in length
Arc-like in shape
Often abraded or tapered at the tips
Pigment in limb hair is generally granular in appearance
Medulla is trace to discontinuous
E. Fringe hair Originating from areas of the body outside those specifically designated as head or
pubic
Not suitable for significant comparison purposes
From the neck, sideburns, abdomen, upper leg, and back.
F. Axillary hair hairs, chest hairs, eye hairs, and nose hairs are not routinely compared

Racial Determination
Caucasoid (European) Mongoloid (Asian) Negroid (African)
Fine to medium coarseness Regularly coarse, straight, Regularly curly or kinky
Appearance: Straight or wavy Cross section: Circular Cross Section: Flattened
Colors: ranging from blonde to Diameter: wider Appearance:
brown to black. Cuticle: Curly, wavy, or coiled
Hair shafts: Significantly thicker Pigment granules:
round to oval in cross section Medulla Larger than those found in
Pigment Granules Fine to Continuous and wider Mongoloid and Caucasian hair
medium-sized, evenly Cortex Grouped in clumps of different
distributed Pigment granules that are larger sizes and shapes.
in size Pigment in the hair shaft so
 great (opaque).

Experiment No. 9
Forensic Analysis of hair
Introduction

Hairs are a potentially ubiquitous trace material in many types of forensic investigation. Few
forensic materials give rise to such differing views on their value as evidence, and these views are often
held with a passion. Some believe that hairs provide worthless evidence, while others believe that hairs
can provide potentially, and actually, very significant evidence. The application of DNA methods to the
examination of hairs has sparked renewed interest in the forensic examination of hairs and the role of
microscopic examination.

Objectives: At the end of this activity, the students are expected to be able to:
a. identify the structure and the characteristics of human hair
b. to determine the differences between hairs from the different regions of the body
c. understand and appreciate the importance of hair analysis in forensic investigation

Materials
a. Hair samples from different parts of the body
b. Hair from different animal sources
c. Reagents: 1:1 mixture of Glycerine and Water, Lacquer
d. Equipment: Glass slide, cover slip, microscope, pincer and Pasteur pipette

Procedure: The preparation of the samples as well as the microscopy will be shown by the instructor
through a recorded video to be uploaded in Canvas.

A. Examine the hair sample and take note of the following: Any foreign materials, color and texture
B. Prepare the Wet mount
a. Wash the hair with soap and water then with alcohol. Dry hair
b. Place the hair on the slide
c. Add a drop of glycerin and water mixture
d. Cover the preparation with a cover glass
C. Examine the mounted hair and observe for the following parts: roots, cuticle, cortex, medulla and
tip

Activity: With the video provided by the instructor and the additional materials from your online and book
references, accomplish the following.

Drawing/Illustration
1. Draw, label and describe the following types of hair.
Scalp Hair Newly cut hair Burnt hair
Eyebrow Hair Freshly pulled hair Torn hair
Pubic Hair Naturally fallen hair Cat hair & Dog/s hair

Questions for Research

1. Can age and sex be identified using hair? Explain how so.
2. Differentiate human hair from an animal hair.
3. Why is DNA analysis in hair samples important?