Chap8 PDF

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
Prepare by: Mark Gideon M. Wallis, RMT
https://www.waynesburg.edu/
Frontiersin.org
Success is no accident. It is hard work, perseverance, learning, studying, sacrifice, and most of all, love of what you
are doing. - Pele
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TABLE OF CONTENTS Page No.
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 3
Study Schedule . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Rubrics for Grading of Activities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
Chapter I. Brief History of Forensic Chemistry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Chapter II. Introduction to Forensic Chemistry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Activity No. 1 Materials and Glass wares in Forensic Toxicology . . . . . . . . . . . . . . . . . . . . 11
Activity No. 2 Instrumentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Activity No. 3 Laboratory Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Chapter III. Blood and Blood stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Activity No. 4 Blood Collection (Venipuncture) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
Activity No. 5 Preliminary test for Blood (ABO blood typing) . . . . . . . . . . . . . . . . . . . . . . . . 25
Activity No. 6 Microscopic Examination for blood . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
Activity No. 7 Microchemical Examination for blood . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Chapter IV. Seminal Fluid Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Activity No. 8 Microchemical Examination if Seminal Fluid . . . . . . . . . . . . . . . . . . . . . . . . . . 34
Chapter V. Hair Analysis in Forensics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
Activity No. 9 Forensic Examination of Hair . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
Chapter VI. Cordage and Fiber Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
Activity No. 10 Microchemical Examination of Fiber . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Chapter VII. Casting and Moulage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
Activity No. 11 Casting and Moulage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
Chapter VIII. Gunshot Residue . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
Activity No. 12 Microscopic and Chemical exam of Gun powder residue . . . . . . . . . . . . . . 54
Chapter IX. Glass and Glass Fracture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Activity No. 13 Reconstructing a Glass Fracture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
Chapter X. DNA Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
Chapter XI. Forensic Toxicology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
Activity No. 14 Isolation and Identification of Poison . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
Chapter XII. Drugs of Abuse . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
Activity No. 15 Microscopic Examination of Marijuana . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
Activity No. 16 Microscopic Examination of Shabu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
Activity No. 17 Drug Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78
Activity No. 18 Microscopic Examination of Barbiturates . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
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Introduction
I. Course Code and Title: FORCHM2 (Forensic Chemistry and Toxicology)
Course Description 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.
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- 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
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Study Schedule
Week Lecture Study Guide Laboratory Study

Number Guide
Week No. 1 CHAPTER I: Brief History of Orientation
Forensic Chemistry & Toxicology
Week No. 2 CHAPTER II: Introduction to Activity # 1
Forensic Chemistry
Week No. 3 CHAPTER III: Blood and Blood Activity # 2 and 3
Stains
Week No. 4 CHAPTER IV: Seminal Fluid Activity # 4, 5 and 6
Analysis
Week No. 5 First Grading Examination
Week No. 6 CHAPTER V: Forensic Analysis of Activity # 7 and 8
Hair
Week No. 7 CHAPTER VI: Cordage and Fiber Activity # 9 and 10
Analysis
Week No. 8 CHAPTER VII: Casting and Moulage Activity # 11 and 12
Week No. 9 CHAPTER VIII: Gunshot Residue Activity #13
Analysis
Week No. 10 Midterms Examination
Week No. 11 CHAPTER IX: Glass and Glass Activity # 14 and 15
Fracture
Week No. 12 CHAPTER X. DNA Typing Activity # 16
Week No. 13 Chapter XI: Forensic Toxicology Activity # 17
Week No. 14 Chapter XII. Dangerous Drugs Activity # 18
Week No. 15 Finals Examination
Note: For offline learners, please feel free to message your instructors through Text
SMS for additional instructions.
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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 point
-Awareness about the specific topic substantial controlling about a specific topic and no to minimal
-Presence of relevant ideas thru point about a specific or question with evidence of
tacts, examples, details, opinions question or topic with sufficient awareness awareness and
and explanation evident awareness and knowledge. (4 knowledge. (1
and knowledge. (7 points) point)
points)
2. Organization 5 Sophisticated Functional Confused or
-Order developed and sustained with arrangement of arrangement of inconsistent
in the paragraph content with evident content that sustains arrangement no
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 grammar
-Choice, use and arrangement of spelling and sentence and minor spelling and major spelling
words formation throughout errors and sentence errors throughout
-Grammar, mechanics, spelling, the paragraph. (3 formation (1 point) the paragraph. (0
usage & sentence formation points) point)
Rubrics for the evaluation of Illustrations and Diagrams

CRITERIA HIGHEST Outstanding Satisfactory Poor
SCORE
1. General 3 Lines or patterns are There are smudges or stray Smudges or stray
Appearance drawn clearly and not pencil marks but these do marks obscure details
- Neatness and smudged. Minimal not significantly affect the of the illustration.
labelling erasure or stray pencil over-all appearance of the Over-all quality of the
marks. Labelling is illustration. Labels are drawing shows
accurate and complete. included but there are minor minimal effort to
(3 points) problems in the accurate complete careful work.
identification of a part. (2 Important labels are
points) missing. (1 point)
2. Organization and 5 Provides complete and Provides clear illustration Only few important
content well-organized however some details are contents are illustrated
-Illustrations and illustrations (5 points) missing (3-4 points) (1 point)
sequencing
3. Attention to details 2 The illustration is Some minor errors are Major discrepancies
accurately drawn (2 present however these do are very noticeable in
points) not distract the information the illustration. (no
conveyed. (1 point) point)
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CHAPTER I: Brief History of Forensic Chemistry & Toxicology
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
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CHAPTER II: Introduction to Forensic Chemistry
Objectives
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
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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. spectrograph – Elemental Analysis
7. Lase Technique – An innovation of spectrographs
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.
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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.
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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.
OBJECTIVES: At the end of the activity, the students will be able to:
a. identify the different laboratory instruments and materials essential in the conduct of Forensic
Chemistry Laboratory;
b. determine the different use/s of each material.
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
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.
ACTIVITIES
Draw and label the parts of the different glass wares, instrument and Materials
QUESTIONS OF RESEARCH
1. Describe each and give the laboratory use or uses of each.
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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).
OBJECTIVES: At the end of the activity, the student will be able to:
a. identify the different laboratory instruments essential in the conduct of forensic toxicology
Laboratory tests;
b. determine the different use/s of the different instruments used in forensic toxicology.
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
PROCEDURE: The instructor will be showing this equipment through a video recording.
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.
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.
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Experiment No. 3
Safety and First-Aid in the Forensic Toxicology Laboratory
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.
OBJECTIVES: At the end of the activity, the student will able to:
a. be familiar with the use of standard precautions in breaking the chain of infection;
b. understand and be able to apply common laboratory safety rules;
c. demonstrate the proper use of personal protective equipment;
d. discuss and be able to apply proper first-aid techniques in response to poisoning
emergencies.
MATERIALS: Reference books

Charts and illustrations
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.
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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.
PROCEDURE
Draw/illustrate the following:
1. Biohazard Sign
2. Safety Signage
3. Personal Protective Equipment
4. NFPA Rating System
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.
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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
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CHAPTER III: Blood and Blood Stains
Objectives
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
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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
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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:
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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.
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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
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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
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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).
OBJECTIVE: At the end of the activity, the student will be able to:
a. know the different methods of blood specimen collection as applied in forensics;

b. perform accurately and precisely the phlebotomy techniques appropriate for forensic
professionals.
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
- Forceps (curve)
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
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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.
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
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.
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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 activity shown by the instructor.
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?
Page 24 of 81
Experiment No. 6
Confirmatory Test for Blood and Blood Stain
Microscopic Examination
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
e. Wright’s stain
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?
Page 25 of 81
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.
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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
Page 27 of 81
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 presents 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
Page 28 of 81
CHAPTER IV: Seminal Fluid Analysis
Objectives
1. assess the importance of seminal fluid analysis in relation to crime investigation
2. schematize the stages of seminal fluid in forensic crime investigation
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/Ejaculate
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
Page 29 of 81
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
Page 30 of 81
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
Materials:
a. Freshly ejaculated 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 eosin
i. Tally counter r. Petroleum jelly
j. Pipettor
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.
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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?
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Assessment No. 3
1. Differentiate Azoospermia 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.
Page 33 of 81
CHAPTER V: Forensic Analysis of Hair
Objectives
1. internalize the importance of hair analysis in crime investigation.
2. differentiate the types and sources of hair
3. criticize the appearance of human from animal hair and from different nationalities.
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
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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
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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
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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
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• 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
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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).
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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 Glycerin 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.
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

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?
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Assessment No. 4
Identify the following:
Identify the source of body Hair

longest hairs on the human body
Arc-like in shape
From the neck, sideburns, abdomen, upper leg, and back.
hairs, chest hairs, eye hairs, and nose hairs are not routinely compared
Called beard hairs or mustache hairs.
Identify the hair as to Racial source
Regularly curly or kinky
Medulla is Continuous and wider
Fine to medium coarseness
Cross section: Circular
Colors: ranging from blonde to brown to black.
Explain briefly
In your own words, how important is Forensic analysis of Hair.
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CHAPTER VI: Cordage and Fiber Analysis
Objectives
1. appreciate the importance of fiber analysis in forensic investigation
2. differentiate the types of fiber investigated in forensics
Introduction
Cordage (rope and string) can be made from many different fibers including (Bast) Dogbane,
Milkweed, Nettles, Hemp, Flax; (Leaves) Cattail, Yucca, Agave, Douglas Iris; (Bark) Willow, Maple,
Basswood, Cedar; (Root) Leather Root, Beach Lupine; (Whole stem) Tule, straw, Juncus.
Trace material such as fibers are fairly unique to an individual’s environment because the choices
in clothing, vehicles and home décor are based on personal preferences. Fibers transferred from an
individual’s personal environment can also be secondarily transferred in a crime. The idea that two
people could be wearing the exact clothing on the same day is unusual therefore the idea that foreign
fibers on a suspect or victim that share the same characteristics after a discriminating analytical
examination is also very unusual.
Ropes and cordage may also be submitted to the Laboratory for analysis. These are either
composed of synthetic or natural fibers. Thus, the same analytical scheme is used as with a fiber
examination. An association between the rope or cordage in question and any known ropes submitted
can be determined.
Fibers can be transferred as easily as they can be lost so lack of evidence does not always
indicate lack of contact. That is why collection and packaging of fiber evidence is important.
Fiber-plastic fusion can occur between the clothing of the occupant and interior surfaces of
the vehicle which are made of thermoplastics. During an impact, parts of the garments can be rubbed
under high pressure against the surface of the vehicle which then causes frictional heat. This, in turn,
causes local melting of the thermoplastic material. During this contact, fibers from the garment can be
transferred and become embedded into the softened plastic. It is the plastic material that is actually
melting and not the fibers.
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Classification of Fibers
I. Natural Fibers
A. Soft Fibers B. Hard Fibers

- obtained from the last layer of the plant stem of - Obtained from the structural system of the leaf of
dicotyledon plants monocotyledon plants
- Examples: - Examples:
Cotton Flax Manila Hemp/Abaca Sisal Hemp
Hemp Jute Coir New Zealand Hemp
II. Synthetic Fibers

Nylon Polyester Polypropylene Aramid
Greatest elasticity Best rope for general Resistant to abrasion High melting point with
use and UV from the sun resistant to stretching
Cordage Fiber Examination
Macroscopic Examination Chemical Examination

a. Check for length and diameter using Vernier Fiber Phloroglucinol Aniline Iodine
caliper sulfate and
b. Examine the general appearance and quality sulfuric
c. Take note of stains, debris, direction of acid
projecting surface fiber Coir Red Yellow Yellow
d. Check for free ends or knots Flax Deep rose Yellow Yellow
e. Check for direction of twist Cannabis Red Violet Yellow Yellow or
-“S” twist or “Z” twist sativa to green
brown
Jute Deep red Bright Brownish
yellow yellow
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Activity No. 10
Fiber Analysis
Micro-Chemical Examination
Introduction
Fiber evidence at a crime scene can be used to link a suspect to a crime. Where the fibers are
found at a crime scene can also suggest the nature of contact that was made by the criminal. For
example, fibers found on a victim might suggest direct contact with that person, while fibers found in a
victim’s apartment may only put the suspect in that location. There are two ways that fibers can be
transferred. Direct transfer also known as primary transfer occurs when the fibers from the suspect’s
clothes comes in contact with the victim. Indirect transfer or secondary transfer takes place when fibers
that are transferred to a suspects clothing are then transferred to the victim.
Objectives: At the end of the activity, students are expected to be able to:
a. describe the different characteristics and/or kinds of fibers
b. determine the importance of fiber analysis in crime investigation
Materials:
a. Wool fibers f. Glass wool
b. Natural silk fiber g. Microscope
c. Artificial silk fiber h. Glass slide
d. Cotton fibers i. Test tube and holder
e. Litmus paper j. Teasing needles
Procedure: The instructor will be recording a video demonstrating the procedures on how to do
fiber analysis. Video will be uploaded in Canvas.
1. Microscopic Examination
- Place the fiber to be examined on a glass slide
- Tease the fiber apart with the use of needle
- Cover with a cover glass
- Observe for the following
o Whether cylindrical, flat or ribbon like
o Whether linen is regular, broad or irregular
o Whether appearing as single thread or paired
o Whether epidermal scale is present
2. Chemical Examination
- Place the fiber to be examined in a test tube
- Heat the test tube until fiber gives off fumes
- Determine the odor of the fumes
- Test the fumes with blue and red litmus paper
- Place the piece of cotton moistened with lead acetic solution and expose this to the
fumes of the burning fiber
- Observe for any blackening of the cotton
- Burn and observe the fiber
Activity:
1. Draw and illustrate the general appearance of the fiber (Wool, Natural silk, Cotton and Artificial
silk)

2. What will be the reaction of the fibers (Wool, Natural silk, Cotton and Artificial silk) to the
following:
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a. Odor of fumes
b. Reaction to litmus paper
c. Reaction to Lead Acetate
d. Rate of burning (Fast or slow)
Page 45 of 81
CHAPTER VII: Casting and Moulage
Objectives
1. appreciate the importance of moulage and casting in crime investigation
2. identify the different use casting and moulage in crime investigation
Introduction
Moulage is a representation of an impression made on a soil surface. A cast in Plaster of Paris or

other similar material of an object or its impressed outlines on a surface. This method has several
importance:
a. Produce representation of an object
b. Great value in scientific criminal investigation
c. Create mold which photography may not become viable
Casting Materials are any materials which can be changed from a plastic or liquid state to the
solid state. The following are criteria for a good casting material:
a. Must be fluid
b. Must harden rapidly
c. Must not be deformable
d. Must be tough
e. Must be easy to apply
f. Must not adhere to the material
g. Must have a fine composition
h. Must not injure the impression
i. Must be obtainable
j. Must be cheap
Materials needed for Casting

1. Impression materials: Clay, Soil, Sand, Snow and Dust
2. Casting Materials
a. Plaster of Paris (CaSO4.10 H20) – Has a poor mechanical strength. Commercially
prepared as Albastone
b. Plastic materials – An example is plasticine
c. Wood’s Metal (Bi.,Pb.,Sn.,Cd.,) – used for small impressions with a melting point at 60-70
degree Celsius but gives more accurate and detailed impression.
d. Negocoll – A rubbery and gelatinous material which is made up of colloidal Mg soaps.
When hot it is thin and pasty, while when cold it is stiff or jelly.
e. Hominit – Made up of resinous material. Flesh in color and used for external surfaces.
f. Celerit – Brown in color and used to cast the human body
3. Molding Box – Holds and keeps casting material from running
4. Reinforcements – includes strip of metals, teased ropes and bamboo sticks
5. Shellac and Alcohol – used as separating materials
6. Talcum powder – used as separating material
7. Chemicals: NaCl, Borax, Sucrose and NaHCO3
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Steps in Casting and Moulage
Page 47 of 81
Activity No. 11
Moulage and Casting
Introduction
Moulage is a representation of an impression made on soil surface and it is the application of a

casting material. A cast of Plaster of Paris or other similar material of an object or its impressed outlines
on a surface to produce representation of an object. It has great value in scientific criminal investigation
and creates mold which photography may not be viable. Always take photograph first before attempting to
make impressions.
a. Appreciate how to do the moulage and casting procedure
b. be able to describe the method of capturing impressions made on a casting material
Materials:
a. Footprint impression (sand) i. Mixing bowl and spatula
b. Hand impression (Clay) j. Water through
c. Frames (shoe box) k. PPE
d. Talcum powder l. Manila paper
e. Shellac
f. Plaster of Paris
g. Pieces of Applicator sticks
h. Hair spray
Procedure: The instructor will be demonstrating through a recorded video the procedure of the
test and will be uploading it in Canvas.
1. Prepare a footprint impression in the sand

2. Surround the impression with hard frames
3. Spray the impression with strong hold hair spray. Spray shellac and let it stand for 5 mins. When
spraying, the sprayer must be held at a distance of about 2 to 3 feet from the impression so as
not to destroy the impression.
4. By means of your hand, mix the plaster of Paris and water on the bowl. See to it that consistency
must not be too thick nor thin.
5. Pour the mixture on top of the impression using your hand as a guide.
6. Cover the impression using your hand as a guide
7. Be very careful that in pouring the plaster of Paris you will not destroy the fine print of the
impression
8. Put pieced of bamboo stick on top of the impression
9. Pour the mixture until you obtain the desired thickness
10. Allow to stand for 30 mins.
11. Repeat the procedure using hand print.
Activity:

1. Aside from foot and hand impressions, what other samples or body parts can be casted?
2. Enumerate common mistakes and errors when doing Casting procedure.
Page 48 of 81
CHAPTER VIII: Gunshot Residue Analysis
Introduction
The primer is detonated when it is crushed by the force of the firing pin. This drives hot gases and
hot particles into the propellant and ignites it. The ignited propellant decomposes and forms gaseous
products. Simultaneously, heat, in enormous quantity, is released by this reaction. The heat generated on
ignition of the primer causes the inorganic ingredients of the primer mixture to vaporize. These vapors re-
condense into droplets, which are further subjected to high pressure and temperature arising from the
openings as vapors and solidified as particulate that varies in shape and size from submicron to over 100
microns. Under ideal circumstances it would be expected that all of the propellant powder would be
consumed in the burning process and would be converted into gases. However, in practice this is not the
case because the whole powder charge is never totally burnt.
Components of Gunshot Residue:

a. Formulation of Primer
b. Formulation of propellent
c. The barrel scrapings
d. The composition of projectile
Ammunition primers consists of four basic chemical components:

1. The initiator – Lead styphnate, is standard initiator in modern primer
2. The oxidizer – Barium nitrate is most commonly used in small arms.
3. The Fuel – Antimony sulfide is commonly used as fuel in primers.
4. The sensitizing – Commonly used sensitizers are tetracene, pentaerythritol tetranitrate and tetryl
Gunpowder: A black powder composed on 75% potassium nitrate, 15% sulfur and 10% charcoal
Combustion product of black powder: Carbonomooxide, carbon dioxide, sulphates, carbonates,

thiosulfate, sulphides and potassium
Smokeless Powder: Can be single based made up of Nitrocellulose or Double base made up of
Mitrpcellulose and Nitroglycerine
Detectable Inorganic components Detectable Organic Components

Lead, Barium, Antimony, Copper from jacket of Nitrocellulose, Nitroglycerine, Diphenylalamine
bullet, Iron from barrel, Nitrates and nitrite, zinc stabilizer, and DNT
and nickel
Location of Gunshot residue: The basic principle of trace and transfer evidence is the Locard Exchange
Principle
Detection of Gunshot residue

1. Residue should initially be observed and evaluated by the unaided eye and with a low power
stereomicroscope
2. X-ray photography
3. IR-photography-Infra red imaging may be used to visualize heavy soot on dark or bloody
clothing or on multicolored surfaces
4. Chemical methods
Collection of Gunshot residue

Collection of Inorganic Gunshot residue Collection of Organic gunshot residue
a. Wet Method: use 5% nitric acid for inorganic a. Swabbing: Most common technique that makes
content use of acetone of ethanol as solvents.
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b. Tape lifting: a technique used to inorganic b. Vacuum lifting: Used for collecting on clothing’s.
sample for SEM analysis Clothing debris are vacuumed on to a filter disc.
Then using solvents, residue on filter paper will be
collected.
Methods of Analysis
1. Gross Examination
- Non-conclusive
- Use of hand lens
- Check for fine black powder particles:
- Entrance of gunshot
- Dorsum of hand
2. Microscopic Examination
- Fine particles magnified
- No characteristic shape, color or consistency of gunpowder
- Paraffin Test
- Diphenylamine Test
- Dermal Nitrate Test
- Lunge Test/ Gonzales Test
- Principle: Nitrates in the gunpowder residue (NO3) reacts with the
diphenylamine
- Positive Result:
1. A deep blue speck developed when NO3 comes in contact with
diphenylamine reagent
- False Negative Result:
1. Use of automatic pistol
2. Direction and wind velocity
- False Positive result
1. Chlorates, dichromates, iodates, bromates, permanganates and higher
metal oxides.
- Walker’s Test
- C-Acid, H-Acid test
- Uses photographic paper treated with either: C-Acid (2-naphthalamine- 4,8-
disulfonic acid), H- Acid (1-amino-8-naphthol-3,6-disulfonic acid) or Sulfonic Acid
swabbed with alpha naphthalamine in ethyl alcohol
- Griess Test
- Principle: Nitrite in acid solution reacts with a primary aromatic amine forming
diazonium salt
1. Sulfanilic acid reacts with nitrous acid to yield a diazonium ion which then
couples with alpha- naphthylamine to produce a red azo dye.
2. Specific for nitrite, not specific for gunshot residues
3. Alpha-naphthylamine a powerful carcinogen
4. N-(1-naphthyl) ethylenediamine is a suitable replacement agent
-
Tests for Primer Components

Methods Principle
Harrison & Gilroy Test Test for Primer components
Sb.,Ba.,Pb.
Swab moistened with 0.1M HCl gathers residues
Neutron Activation Ba and Sb converted to isotopes by neutron bombardment

Analysis (NAA)
Flameless Atomic High temperature vaporizes metallic elements
Absorption Detected through Absorption Spectrophotometry
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Spectrophotometry
(FAAS)
Scanning Electron Adhesive to gather any particles
Microscope w/ X-Ray Analyzed through Scanning Electron Microscopy
Analyzer X-Ray Analyzer confirms their presence
HPLC Analyze pre-cleaned hand swab

extracts obtained from hands after firing a revolver
Pendant Mercury Drop Electrode (PMDE) Detector
Detect down to 1 ng/swab of nitroglycerine.
Fluorometric Detection One of the most sensitive
Selective modes of detection
Organic constituents of gunshot residues identified by molecular
luminescence.
Supercritical Fluid The mobile phase is a supercritical fluid such as CO2, N2O, NH3, or SF6.
Chromatography (SFC) Solvation power for SF and higher separation efficiency in SFC
Capillary Provide rapid, high-resolution separations of complex mixtures

Electrophoresis
Page 51 of 81
Activity No.12
Forensic Analysis of Gunshot Residue
Microscopic and Chemical Analysis
Introduction
When a gun is fired, hot gasses and small particulates are generated during explosion of the
primer and the propellant in the cartridge casing. These particulates, as well as those formed by the
condensation of some of the hot gases are deposited on parts of the hand and clothes. Of the person
firing the gun as well as on other objects in the vicinity of the weapon. These small particulates are called
“Gunshot Residues: They have distinctive characteristics and can be analyzed by several method.
Samples of Gunshot residues are collected from people suspected of firing a gun by pressing adhesive
material to the area of interest, for example, fingers, the top of the hand and the clothes.
Paraffin test is one of the crudest methods for detecting the presence of gunshot residue. This
method detects the presence of nitrate residue. In this test warm paraffin, which is applied to the skin to
open up its pores, collects contaminants. If the suspect had fired a gun, one potential contaminant would
be the nitrates from gun powder residue. Once the paraffin hardens, either diphenylamine or diphenyl
benzidine is introduced to the paraffin cast, which will turn it blue in the presence of nitrates. Thus, the
presence of blue dots on the paraffin casts is evidence that the suspect had fired a gun.
a. appreciate how to detect and identify the microscopic appearance of a gunshot residue
b. appreciate the proper way of how to identify the presence of nitrate in gun powder residue using
paraffin cast technique
c. determine the importance of gunshot residue analysis in identifying suspects firing a gun as well
as other objects in the vicinity of the weapon.
Materials:
a. Compound microscope e. Cotton gauze
b. Cotton swab f. Beaker
c. Magnifying glass g. Tong
d. Transparent adhesive tape h. manila paper
e. Bunsen burner i. Nitric Acid
f. Tripod j. Freshly prepared diphenylalanine
g. Wire gauze
h. Paraffin wax
Procedure: The instructor will be recording a video demonstrating the proper way of analyzing gunshot
residue using microscopic analysis and paraffin technique. The results will be shown and explained by
the instructor.
A. Microscopic Examination of Gunshot Residue

a. Inspect the subject hand using a magnifying glass of any gunshot residue
b. Moisten the cotton swab with 1M Nitric acid
c. Swab the back of the firing hand and the thumb area
d. Use the adhesive tape to blot the skin previously exposed with 1M nitric acid
e. Run the length of the adhesive tape in the surface of the slide
f. Mount under the microscope and examine the sample under HPO
B. Paraffin test or Lunges Diphenylalanine test

The test is conducted by applying melted paraffin wax to the back of a suspect's hands. With a
brush, the back of the hand is coated with paraffin wax which on cooling solidifies and can be peeled off
the hand. The surface of the cast that has been in contact with the skin is treated with diphenyl-
amine/sulfuric acid reagent by dropwise addition or spraying. The reagent produces a blue color with
individual particles of nitrates and nitrites.
Page 52 of 81
Activity:
1. Enumerate and explain the methods of how to collect gunshot residue samples.
2. Explain the principle of the paraffin test.
3. Is the paraffin test still used today?
4. Aside from what was performed in the lab, enumerate and explain two more test to detect the
presence of gunshot residue.
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CHAPTER IX: Glass and Glass Fracture
Introduction
Glass is an important physical evidence because it breaks and pieces are scattered at the crime
scene and on a suspect. It is nearly impossible for any suspect involved in a scene where it is present not
to take away a trace. A glass impact will shower fragments into a surprisingly large radius – up to around
three meters. If a forensic investigator trained in glass recovery manages to trace an intruder and seize
some clothes – if he was wearing them at the time – there’s a significant chance it will have glass that
would tie him to the scene in question. Glass can also be found in some unlikely places too – like
our burglar’s hair, ears, underneath the fingernails or even in the skin. Investigators are called upon to
determine whether a window was broken from inside or outside. Determine which of two holes in a
window was made first. A hole was caused by stone, bullet or some other forces.
Objectives
1. justify the use of glass fractures in crime investigation.
2. appreciate the importance of glass fracture reconstruction in forensic crime investigation
Characteristics of Glass:
a. Hard non-crystalline material
b. Usually clear and transparent
c. Glassy state or vitreous state
d. Atoms arranged at random rather than regular
e. Fluid at high temperature
f. Super cooled liquid
Composition of Glass: Sand (silica), Soda, Lime and other trace elements
Additive’s responsibilities:
a. Alumina – Aluminum oxide
- Units formed lead to improved chemical durability and viscosity
b. Boron-Oxide – Very resistant to heat
- Addition used in borosilicate & aluminoborosilicate glasses.
c. Soda Ash – lower silica’s melting point
- Allows metal containers for processing (fluxing agent)
d. Glauber’s salt
- Sometimes used instead of soda ash
e. Lime
- Added to improve hardness & chemical durability
f. Lead oxide – high lead content lowers melting point
Types of Glass
a. Aluminosilicate & Borosilicate
b. Laminated Glass
c. Lead Glass
d. Soda lime Glass
e. Tempered (stressed) glass
f. Potash
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Methods of Glass Analysis
Spectrograph X-ray analysis Physical properties UV Examination

The only adequate method Study of Most sensitive method in Examined in a dark room
for chemical analysis diffraction determining differences A difference in fluorescence
Not rapid method pattern in composition of glass is indicative of physical and
chemical differences
Glass as evidence in Crime, Forensic chemistry application:

a. Automobile glass in cases of hit and run
b. Broken windows caused by pressure, blow or bullets
2 Kinds of Glass Fractures
Radial Fractures Concentric Fractures
Primary fractures Secondary Fractures

Resemble spokes of wheel Circles around the point of impact
Radiating outward from the point of impact Connecting to one radiating crack to others
Breaking of Glass
When a force pushes on one side of a plane of glass,

the elasticity of glass permits it to bend in the direction of the
force applied. Once the elastic limit is exceeded,
the glass begins to crack.
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3 R’s Rule
Radial cracks form a Right angle on the Reverse side of the force. Radial cracks are first
commencing on the side of the glass opposite to the destructive force. Stress lines on a concentric crack
will be at right angles to the front. Concentric cracks occur afterwards. Starting on the same side as the
line force.
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Activity No. 13
Glass Fragments and Fracture
Introduction
Glass is such a common material that it shows up on soles and clothing for almost anyone with
so much glass picked up from casual contact, courts and investigators needs someway to determine
which fragments actually have relevance in cases. This makes forensic glass analysis essential to the
criminal justice.
Objectives: At the end of the activity, the students are expected to be able to appreciate how to
determine and identify the direction and the means a glass pane was broken.
Materials:
a. Window glass
b. Illustration board
c. Hand lens
d. Broad scotch tape
e. Hammer
Procedure: The instructor will be demonstrating through a recorded video the procedure of the test and
will be uploading it in Canvas.
A. Direction of Force
a. Break a piece of glass by means of a hammer
b. Collect and reconstruct the broken pieces of glass
c. Examine the cross section and striation found on the edge of each
d. Determine the first and radial cracks
e. Determines the stress lines
f. Determine the direction where force was applied from the result of your examination.
B. Reconstruction of the broken glass
Activity:

1. Discuss how do you establish the direction of force applied in the glass.
2. Describe the following method of analyzing glass
a. Floatation method
b. Immersion method
c. Beckeline method
Page 57 of 81
CHAPTER X. DNA Typing
Objectives
1. discuss the importance of DNA typing in relation to crime investigation
2. appreciate the different practical uses of DNA
Introduction
"DNA typing" is a catch-all term for a wide range of methods for studying genetic variations. Each
method has its own advantages and limitations, and each is at a different state of technical development.
Each DNA typing method involves three steps:
1.Laboratory analysis of samples to determine their genetic-marker types at multiple sites of potential
variation.
2.Comparison of the genetic-marker types of the samples to determine whether the types match and
thus whether the samples could have come from the same source.
3.If the types match, statistical analysis of the population frequency of the types to determine the
probability that such a match might have been observed by chance in a comparison of samples from
different persons
What is a DNA?
DNA, or deoxyribonucleic acid, is the hereditary material in humans and almost all other
organisms. Nearly every cell in a person’s body has the same DNA. Most DNA is located in the cell
nucleus (where it is called nuclear DNA), but a small amount of DNA can also be found in the
mitochondria (where it is called mitochondrial DNA or mtDNA).
The information in DNA is stored as a code made up of four chemical bases: adenine (A),
guanine (G), cytosine (C), and thymine (T). Human DNA consists of about 3 billion bases, and more than
99 percent of those bases are the same in all people. The order, or sequence, of these bases determines
the information available for building and maintaining an organism, similar to the way in which letters of
the alphabet appear in a certain order to form words and sentences.
DNA bases pair up with each other, A with T and C with G, to form units called base pairs. Each
base is also attached to a sugar molecule and a phosphate molecule. Together, a base, sugar, and
phosphate are called a nucleotide. Nucleotides are arranged in two long strands that form a spiral called
a double helix. The structure of the double helix is somewhat like a ladder, with the base pairs forming the
ladder’s rungs and the sugar and phosphate molecules forming the vertical sidepieces of the ladder.
An important property of DNA is that it can replicate, or make copies of itself. Each strand of DNA
in the double helix can serve as a pattern for duplicating the sequence of bases. This is critical when cells
divide because each new cell needs to have an exact copy of the DNA present in the old cell.
What is DNA Typing?
Page 58 of 81
This is a process where in DNA extracted from a biological sample is tested. The DNA is
processed to provide identification of genetic sequences.
Page 59 of 81
Chapter XI: Forensic Toxicology
Objectives
1. evaluate the importance of poison isolation in forensics
2. Determine the different examples of dangerous drugs
Introduction
Toxicology is branch of science which treats poisons, their origin, physical and chemical
properties, physiological action, treatment of their noxious effect, and their methods of detection. Poison
is any substance which produces deleterious effects of the living tissues and once introduced into the
blood stream even in a small dose, is likely to cause injurious effect to health and even cause death.
Types of poisoning
Accidental Poisoning Taken without the intention to cause death
Suicidal Poisoning Taken voluntarily for the purpose of taking one owns life
Homicidal Poisoning Given willfully with intent to cause death to the victim
Undetermined Poisoning Problem how poison was obtained and why it was
administered
Conditions Modifying Actions of Poisons
A. Attributed to individuals
a. Age and Sex e. Food
b. Habit f. Disease
c. Health g. Sleep
d. Idiosyncrasy h. Exhaustion
B. Attributed to the poison

Dilution More dilute, rapidly absorbed except corrosive poison
Dose Quantity of poison administered at a time
Physical State Gaseous is more absorbed than solids
Solubility Must be capable of solution to be absorbed
Mode of Rapid action when inhaled in aqueous or vapor form
Administration
Chemical When compared with others, more poisonous
Combination
Mechanical Medium where poisons are dissolved
Combination
Application of Poison Absorption is rapid in blood
Site of Entry of Poisons

a. Mouth f. Urethra
b. Nose g. Bladder
c. Eyes h. Ureter
d. Rectum i. Through injection
e. Vagina
Poison Elimination
a. Emesis e. Milk
b. Respiration f. Sweat
Page 60 of 81
c. Feces/Stool g. Saliva
d. Urine h. Tears
Types of Poisons
Organic poisons Synthetic drugs, Alcohols, Petroleum product, Insecticides,
Chlorocompounds, Aromatic and Organo-P-compounds
Inorganic poisons Metals, metalloids, non-metals, salts, acids, bases, As, Sb, Pb,
Salts, Cyanides, Fluorides, CO, Halogens
Biological poisons All poisonous plants and their parts and extract. Opium, cocaine
and atrophine
Collection of Samples
a. Vomitus
b. Purged materials
c. Urine or fecal stains
d. Clothes, bed sheets and bed cover
e. Medicines the victim or family have been using
f. All containers of medicine
g. Remains of food and drinks
h. Containers of food and drink
i. Cooking utensils
j. Solids and liquids contained in traps of sink
Autopsy Materials
The following materials are a MUST
a. Vomitus and purges e. Intestines (whole)
b. Blood (500 mL) f. Kidneys
c. Urine (Total amount recovered) g. Spleen
d. Stomach contents
Symptoms of Poisoning
a. Vomiting – due to As, acids, alkali, excess liquor and metallic salt
b. Diarrhea – same agents as A
c. Cramps – As, Sb and Pb
d. Delirium – Diarrhea, Cannabis, alcohol, atropine, Hyoscine, LSD
e. Convulsion – Nicotine, Cyanide
f. Paralysis – Snake venom, acotine and As
g. Coma – Barbiturates, opium, CO, Chloroform, Excess liquor
Preliminary test for Poisons
Microscopic Examination
a. Physical State
b. Color, Odor
c. Mixtures of materials
Reaction with Filter Paper

a. Acid Reaction suggestive of the presence of free acids and acid salts or readily
hydrolyzed salts
b. Alkaline Reaction suggestive of the presence of caustic alkalis of ammonia
Page 61 of 81
Flame Tests (Beilten Test)
a. Presence of organic compound is suspected to contain halogens.
b. Moistening a copper wire with the poison
Luminosity Test
a. When Phosphorous is suspected.
b. Small quantity of material is placed in a test tube, acidified with dilute H2SO4.
c. Shaken and heated between 40- 50’C.
Short cut Methods of Toxicological Analysis

a. Reinsch’s Test
- Sample is heated with water strongly acidified with HCl.
- Chemical copper coil or wire is placed in the mixture
b. Mitscherlich’s Test
- Determines Phosphorous from the sample
- Organs acidified with dilute H2SO4
- Collected in dilute HNO3
Page 62 of 81
Experiment No. 14
PREPARATION OF APPARATUS USED
IN THE ISOLATION AND IDENTIFICATION OF POISON
(TOXICOLOGY)
INTRODUCTION:
Forensic toxicology is an examination of all areas of toxicity to aid medico-legal enquiries.

Toxicology is the study of poisons. It takes into account the origins and properties of the poisons;
including chemical and physical properties and their effects on living organisms as well as the corrective
measures for the treatment of poisoning. A forensic toxicologist is interested largely with exposure and
evaluation of poisons in tissues and body fluid acquired at autopsy or a living person in blood, urine or
gastric material. Steam distillation works on the principle that immiscible objects when mixed together can
lower the boiling point of each other. Historically and in some regions of the world today essential oils
were obtained by a rather primitive and ugly process. By boiling some organic matter with water and then
condensing the resultant vapor /gas, one can obtain some essential oils. (http://www.ukessays.com /
essays/biology/).
OBJECTIVE: At the end of the activity, the students are expected to be able to appreciate the preparation
of the different set-ups in the isolation and identification of poisons.
MATERIALS
Bunsen Burner Florence Flask

Condenser Cork Borer
Wire Gauze Rubber Tubing
Glass Tubing File
Distilling Flask Iron Stand (2)
Iron Clamp (2) Beaker (200 ml)
Sterile Urine Container (2) Gloves
Mask
PROCEDURE: The instructor will be demonstrating through a recorded video the procedure of the test
and will be uploading it in Canvas.
Distillation Set-Up: The proper way of setting up will be shown by the instructor.
1. Examine the illustration of the Distillation Set-up.
2. Study the diagram provided and carefully prepare the Steam Distillation set up.
3. The setup of instruments will be checked by the instructor before operating the
system.
Page 63 of 81
http://ph.images.search.yahoo.com/search/images
4. Combine equal parts (30 ml) of 95% ethyl and urine sample and dispense in
the distilling flask. (instructor to explain the proper technique)
5. Assemble the thermometer in a tightly fitted cork. The bulb of the thermometer
should be submerged in the solution.
6. Boil the solution and maintain a temperature lower than the boiling point of an
ethyl alcohol throughout the process.
7. Collect the distillate using a beaker (30 ml) and transfer in s sterile urine
container.
8. Keep the distillate for the next laboratory activity.
1. What are other methods used in forensic toxicology to separate the poisons from the entire
specimen? Describe each.
2. Discuss the principle of separation in the following processes:
a. Gel electrophoresis
b. Dialysis
c. Chromatography
d. Precipitation
e. Sedimentation
3. What is solvent extraction? Discuss completely
Page 64 of 81
Chapter XII. Dangerous Drugs
Objectives
1. differentiate dangerous from prohibited drugs
2. identify the different features of dangerous drugs
Republic Act No. 9165 June 7, 2002, This Act shall be known and cited as the
"Comprehensive Dangerous Drugs Act of 2002". It is the policy of the State to safeguard the
integrity of its territory and the well-being of its citizenry particularly the youth, from the harmful
effects of dangerous drugs on their physical and mental well-being, and to defend the same against
acts or omissions detrimental to their development and preservation. In view of the foregoing, the
State needs to enhance further the efficacy of the law against dangerous drugs, it being one of
today's more serious social ills.
At the end of this chapter, you should be able to:

a. Discuss the mechanism of action of various toxins in the body.
b. Associate clinical signs & symptoms to particular types of toxicity/poisoning.
c. Describe the effects of varying blood alcohol concentrations on behavior.
d. Describe the methods employed for alcohol determination.
e. Classify the drugs of abuse as to mode action
f. Describe the techniques for drug analysis.
g. Accurately perform a screening drug test following the proper protocol.
Section 1. DRUGS OF ABUSE
Nature of Drugs and Drug abuse
Drugs are any substance that produces physiological or psychological change within a short
period of time after ingestion of a specified dose. Different types of drugs affect your body in different
ways, and the effects associated with drugs can vary from person to person. How a drug effects an
individual is dependent on a variety of factors including body size, general health, the amount and
strength of the drug, and whether any other drugs are in the system at the same time.
Nature of Drug dependence
When an individual becomes strongly attached to a drug. Dependency is subdivided into two
categories: physiological and psychological. Drugs can have short-term and long-term effects. These
effects can be physical and psychological, and can include dependency. Psychological dependence is
when a person develops an uncontrollable “craving” (mental or emotional need) for a drug, the craving is
a desperate need to continue. In physiological dependency, the body continually needs to have the drug,
a person experience sickness if drug is discontinued.
Six Categories of Drugs of abuse
A. Opiates and Narcotic B. Stimulants C. Hallucinogens

Drugs
*Morphine is the primary *Stimulants are taken to *Cause a significantly altered mental state, often
active drug in opium that make one feel more including hallucinations
came from the dried sap energetic, strong, or awake
of the opium poppy plant. *Marijuana is one of the oldest
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*Abused stimulants: *Physiologically active ingredients: Cannabinoids,
Amphetamine, found in the resinous leaf coating of Cannabis sativa
methamphetamine, and
cocaine *The most active cannabinoid
is THC
Livescience.com *Methamphetamine is the
drug most commonly
*Opium can be smoked produced in clandestine labs
directly or chemically
processed to isolate pure
morphine
*Opiates are Medicalnewstoday.com

psychologically addictive
drugs; withdrawal causes Livescience.com
*Cocaine - very powerful
severe physiological
stimulant; enormously
symptoms
psychologically addicting *Metabolites of Marijuana:
*Codeine - second most
*Cocaine hydrochloride is -Delta-9-carboxy-THC
abundant component of
usually inhaled through the
opium, used as a strong -111-hydroxy-delta-9-THC
nose
painkiller and cough
suppressant *Its free base form (“crack”) -Detectable in urine from 1 to 4 weeks after last use
is vaporized by heat in a
*Hashish – more potent form of marijuana made
pipe and inhaled into the
from the flowering tops of the plant
lungs
*Hash oil: made by heating the plant material in a
solvent, then evaporated leaving a thick oily material
(almost pure resin)
Webmd.com *Hash oil can be mixed with tobacco or other

Sunrisehouse.com vegetable material
*LSD is an extremely potent hallucinogen with

normal dose of 30-50 mg, this causes visual
hallucinations, brilliant colors and the perception that
one is wise
Medicalexpress.com
Page 66 of 81
*Naturally occurring hallucinogens: Peyote, the bud
of a particular cactus whose main ingredient is
mescalin
*Magic mushrooms: Genus Psilocybe; Active

components: psilocin and psilocybin
e.wikipedia.org
D. Depressants, Hypnotics and E. Club Drugs F. Athletic Performance

Tranquilizers Enhancers
*Alcohol (depressant) - most abused *prepared by clandestine labs, or obtained *Athletes trying to gain a
substance legally from other countries competitive edge may
abuse stimulants and
*MDMA - “love drug” or “Ecstasy” painkillers
*GHB, gamma hydroxybutyrate, *The first drug controlled

because of their abuse
*GHB and related compound GBL:
by athletes were anabolic
used for their hypnotic or depressant steroids
Laressio.com
effects
*Anabolic steroids
promote cell growth
resulting in growth of
*Barbiturates - physiologically
muscle tissue and
active depressants, resulting in a sometimes bone size and
strength
physical & mental state similar to Theconversation.com
Page 67 of 81
alcohol-induced intoxication *Ketamine - anesthetic and animal
tranquilizer; causes anterograde amnesia
*Rohypnol, GHB, and ketamine have been

implicated in cases of drug-facilitated
sexual assaults “date-rape” drugs
Drugtestsinbulk.com
*Condensation product of urea and

malonic acid En.wiki.com
*Fat soluble : easily crosses BBB
*Low doses: sedation, drowsiness and

sleep
*Higher doses : anesthesia

Statnews.com
*Very high doses : stupor, coma and
death
*Toxicity : depression, cyanosis,

hypothermia, hypotension
*Valium (a benzodiazepine) - a
tranquilizer drug designed to relieve
anxiety
*Rohypnol or “roofies” is a
benzodiazepine and a major drug of
abuse at raves and
the club scene
Testing for Drugs of Abuse
Drug testing centers over the years had an increasing capability. Drug testing has been required
by government agencies, industrial agencies and sports agencies. Forensic drug testing has been used
for the application of drug test in questions of law. The following are specimen requirements and the
different methods used for drug testing:
• Urine specimen – the usual sample used to test a number of drugs of abuse
o Dias advantage:
▪ Detects only fairly recent drug use
▪ Will not differentiate casual use from chronic drug abuse
▪ Does not determine degree of impairment
▪ Does not determine dose of drug taken
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▪ Will not state exact time of use
• Meconium
o first stool of the newborn
o begins to form during second trimester and continues to accumulate until birth
o provides evidence of maternal drug use anytime during the last two trimesters
• Hair
o obtained easily; not easily tampered
o prior drug use may be detected for several months
o hair growth: 0.3 to 0.4 mm/day
o mechanism of drug deposition in hair
▪ transfer from blood to growing hair shaft
▪ transfer from sweat
▪ environmental contamination
• Sweat
o sweat-patch collection devices, worn for several days to several weeks
o the drug, if present accumulates on the absorbent pad in the patch
o advantage: monitoring of drug use in correctional institutions or in drug rehab programs
• Saliva
o easy to obtain, less invasion of privacy and ease of adulteration
o ultrafiltrate of plasma and used for recent drug use
o detection of drug level is shorter compared with urine
Laboratory Considerations
Specimen collection
o urine: sample of choice; represents the net load of the drug over a long period
o guard against specimen exchange and/ or adulteration
o urine pH, specific gravity and creatinine
o blood: represents transient passage of the drug thru the circulation
o only a quick picture of the drug level at a specific time
Processing and handling – confidentiality and chain of custody
Analytical methods
a. screening assay – good sensitivity with marginal specificity
b. confirmatory assay – high sensitivity and specificity
- qualitative and quantitative info
- different from screening procedure
Techniques for Drug Analysis

 IMMUNOLOGIC
I. Enzyme Multiplied Immunoassay Technique (EMIT)
o Drug to be measured: hapten part of an antigen
o Serum + antibody mixture + enzyme labeled drug + substrate
o Measure enzyme activity: drug concentration
o More rapid than RIA
II. Enzyme-Linked Immunosorbent Assay (ELISA)

o Drug to be measured is the hapten
o Specific antibodies bound to a solid-state carrier
o Separation of the bound drug from the unbound
III. Fluorescence Immunoassay

o Drug bound to a fluorogenic substrate (Umbelliferyl-β-galactoside)
o Fluorogenic drug reagent + antibody + beta-galactosidase --- incubated with serum
sample
o Drug and FDR compete for binding
o Fluorescent product = Umbelliferone
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IV. Radioimmunoassay
o Drug (hapten) + protein + antibody
o Incubation of serum + antibody + radiolabeled drug (competition for antibody binding
sites)
o 2 fractions: Free fraction (unbound drug) and Antibody-bound drug
 CHROMATOGRAPHIC
I. Chromatography
o Adsorption of drug to a solid support and elution by means of a mobile , liquid phase
o TLC – screening for drug identification
Thin Layer Chromatography
o use solid phase support medium and liquid mobile phase separation system
o drugs are separated on the basis of their ability to dissolve in the solvent system and its
strength of interaction with the support phase
o color reactions are then used to locate and identify the specific substance
o urine sample adjusted to pH 8.5
HPLC and GLC- primarily for quantitating serum drug levels in TDM and also for confirming drug
identification
Gas Chromatography/ Mass Spectrometry
o most specific and sensitive: gold standard
o parent drug and metabolites may be detected
o sample is placed in a solvent to extract the substance
o extract is concentrated and injected into a gas chromatograph
o fractionation pattern is determined
o mass spectrometry: determine molecular weight and structural characteristics to identify
the drug
 SPECTROPHOTOMETRIC
o Spectral scan: tentative identification of drugs
o Protein precipitation or extraction
o Quantitative analysis
Section 2. TOXIC AGENTS

• Routes of exposure: ingestion, inhalation and transdermal absorption
• Absorption in the GIT is by passive diffusion: this process requires that the substance cross
cellular barriers
• Toxins that are not absorbed from the GIT do not produce systemic effects but may produce local
effects – diarrhea, bleeding and malabsorption of nutrients.
A. Alcohols - common CNS depressants

• Cause disorientation, euphoria, confusion and may progress to unconsciousness, paralysis and
even death
1. Ethanol
▪ Most commonly abused depressant
▪ Toxic metabolite: acetaldehyde
▪ Increase in GGT, AST, AST/ALT ration (>2.0), increased HDL and MCV
▪ Serum, plasma and whole blood are acceptable specimen (venipuncture site should be
cleaned with alcohol-free disinfectant – benzalkonium chloride)
▪ The specimen must be capped all the time to avoid evaporation of alcohol
▪ Methods for testing: enzymatic, GC and osmometry
▪ Detection limit: 12 hours
▪ Fatal dose: 300 – 400 mL of pure alcohol consumed in <1 hour
Page 70 of 81
Influence of Acute Ethanol Ingestion on Ethanol Levels and Behavior
BAC (mg/dL) Influence
10-50 mg/dL None to mild euphoria
50-100 mg/dL Mild influence on stereoscopic vision and dark adaptation
100-150 mg/dL Euphoria; disappearance of inhibition; prolonged reaction time
150-200 mg/dL Moderately severe poisoning; reaction time greatly prolonged;

loss of inhibition and slight disturbances in equilibrium and
coordination
200-250 mg/dL Severe degree of poisoning; disturbances of equilibrium and

coordination; retardation of thought processes and clouding of
consciousness
250-400 mg/dL Deep, possibly fatal coma
(lifted from Henry’s Clinical Diagnosis and Management by Laboratory Methods, edited by
McPherson and Pincus)
Stages of Acute Alcoholic Influence / Intoxication

BAC (g/100 mL) INFLUENCE
0.01 – 0.05 Subclinical
0.03 – 0.12 Euphoria
0.09 – 0.25 Excitement
0.18 – 0.30 Confusion
0.25 – 0.40 Stupor
0.35 – 0.50 Coma
0.45+ Death
Modified from Dubowski KM, Gadsden RH Sr, Poklis A. The stability of ethanol in human whole blood
controls: an interlaboratory evaluation. J Anal Toxicol. 1997 Oct;21(6):486-91. All rights reserved
(lifted from Tietz Textbook of Clinical Chemistry and Molecular Diagnostics 5 th edition by Burtis, Ashwood
& Bruns)
OTHER ALCOHOLS
NAME/SYNONYMS SOURCES TOXIC SYMPTOMS OF METHODS FOR
METABOLITE TOXICITY DETERMINATION
Methanol / wood Methylated Formic acid Ocular toxicity, GC-MS

alcohol spirits metabolic acidosis
Anti-freeze
agents
Page 71 of 81
Isopropyl alcohol / Antiseptic Acetone Acetonemia, acetonuria Gas
rubbing alcohol agent & hyperosmolarity Chromatography
without hyperglycemia
& acidosis
Ethylene glycol / Hydraulic fluic Glycolic acid Deposition of calcium HPLC

1,2-ethanediol & antifreeze oxalate crystals in renal
agent Oxalic acid tubules
Assessment No. 5
Question: On the space provided, differentiate a screening from a confirmatory drug testing
laboratory as to service capabilities and test processes.
Question: In your opinion, was the War on Drugs by the President successful?
Page 72 of 81
Reference:
Hayes, Wallace A (2007). Principles and Methods of Toxicology (5th ed.). USA: Informa
Healthcare.
Katzung, B.G., ed. (2009) Basic and clinical pharmacology. (11th ed.). Boston : McGraw-Hill.
Page 73 of 81
Experiment No. 15
APPLICATION OF MICROSCOPIC EXAMINATION
ON MARIJUANA
INTRODUCTION:
A very small amount of the dried unknown is selected. This becomes the sample. The sample is
placed on a microscopic slide. A drop or two of water is then added to the slide. The slides are examined
at varying levels of magnification and under different light conditions. What the analyst is looking for is
two distinct morphological features. They are looking for microscopic “hairs” on the unknown. These are
cystolithic hairs and glandular hairs. Cystolithic hairs are often likened to like little bear claws in their
appearance. Some techniques call for the use of hydrochloric acid after they look for these hairs. A few
drops of HCL are added by the analyst. The analyst then looks to see if there is some unspecified
effervescence under the light of the microscope
(http://www.thetruthaboutforensicscience.com/tag/microscopic-morphological-examination-for-
marijuana/).
OBJECTIVE: At the end of the activity, the students are expected to be able to:
a. identify the microscopic characteristics of marijuana;
b. determine the morphologic features of marijuana.
MATERIALS AND REAGENTS
Samples
Marijuana leaves and seeds, papaya leaves, shredded tobacco (cigarette of any brand).
Tobaco Americano or any available leaves resembling marijuana in form.
Materials and Instruments

Cover Slip
Glass slides
Electric Microscope
Reagent
Dilute HCl
1. Place few pieces of crushed leaves suspected to be marijuana on a glass slide
and view the preparation under the microscope (LPO).
2. Observe the fine hairs that appear like bear claws and the spheroidal crystolith
on its base.
3. Place a drop of dilute HCl against the specimen after the microscopic
examination.
4. Take note of the effervescence.
5. Repeat the above procedure using other samples.
1. Describe marijuana as follows:

a. RA 9165
b. Pharmacological Classification
c. 3 components (describe)
d. Drug schedule
e. Signs or symptoms of marijuana intoxication
f. Other names
Page 74 of 81
g. Route of introduction
h. Use/uses
2. What are the chemical tests for marijuana? Describe each and give the positive results.
3. What is medical marijuana? Discuss completely.
4. Are you in favor of medical marijuana? Explain your
Page 75 of 81
Experiment No. 16
ON SHABU (Microcrystal Test)
INTRODUCTION:
Shabu is an amphetamine derivative (trade name Methedrine) used in the form of a crystalline
hydrochloride; used as a stimulant to the nervous system and as an appetite suppressant. Shabu is a
prohibited drug. Said to be addictive and destructive. For occasional users l, here are the effects such as
Insomnia which can last for as long as three days, feeling energized and alert, increased activity,
increased sexuality and makes you lose your appetite (answers.com/Q/What
_is_shabu_and_what_is_the_effect_in_the_body).
OBJECTIVE: At the end of the activity, the students are expected to be able to:
a. identify the presence of methamphetamine (shabu) using the microcrystal test;
b. determine the physical and chemical characteristics of shabu.
a. Cavity Slide Reagents

b. Pasteur Pipette - 5% NaOH Solution
c. Glass Slide - 5% HAuCl in H3PO4 (Gold Chloride)
d. Electric Microscope
Procedure: The instructor will be demonstrating through a recorded video the procedure of the test and
will be uploading it in Canvas.
A. Preparation of Reagent
5% HAuCl in H3PO4
- Dissolve 1 gram of commercial gold trichloride acid (HAuCl4 x 4H20) in
20 ml of a solution containing 1 volume of concentrated H3PO4 and 2
volumes of water.
B. Procedure
1. Transfer a small quantity of the sample powder into the depression of
the cavity slide.
2. Add a drop. of the volatizing reagent (5% NaOH solution).
3. Immediately transfer a drop of testing reagent (5% HAuCl in H3PO4)
4. Place a cover slip on top of the sample cavity and stand for 10 minutes at
room temperature.
5. After 10, minutes, examine the crystals in the reagent or at the edge of the
reagent drop under the microscope using the LPO.
7. Illustrate and describe result.
V. QUESTIONS FOR RESEARCH
1. Describe shabu based on the following parameters.

a. RA 9165
b. Pharmacological Classification
c. Active components
d. Route of administration
2. Signs of shabu intoxication
3. What are the chemical tests used in the detection of shabu?
Page 76 of 81
Experiment No. 17
Drug Testing: MET and THC
OBJECTIVE: At the end of the activity, the students are expected:

a. To properly perform the screening test for drugs of abuse based on Thin Layer Chromatography
b. To describe the principle underlying Thin Layer Chromatography as a screening test for drugs of
abuse
c. To appreciate the significance of monitoring “chain of custody” in drug testing
MATERIALS:
• Specimen: Urine
• Drug Testing kits
PROCEDURE: The instructor will make a video of the actual laboratory performance of the procedure
and explanation of the principle of the tests. The video will be uploaded for the students to be able to
familiarize themselves with the procedure. The video will explain the complete process of screening drug
tests and the results of such test.
GUIDE QUESTIONS: The instructor will make an online forum where the students can type and post their
answers to the QFRs. For offline students, they can write it on an intermediate pad to be submitted via
mail, express couriers or through text message.
1. Discuss the principle associated with screening drug testing (TLC) and confirmatory drug testing
(GC-MS)
2. In tabular form, characterize the different drugs of abuse based on:
a. Other names (common names/chemical name)
b. Mode of action
c. Detection Limit & Detection Time in Urine
3. Discuss the concept of “Chain of Custody” in drug testing.
Page 77 of 81
Experiment No. 18
ON BARBITURATES
INTRODUCTION:
Forensic drug analysis deals with the identification and quantification of illegal drugs. Forensic
drug tests are generally carried out in two steps: screening and confirmation. Once drugs are detected
through screening, for example spot test kits (e.g., immunoassays, Marquis test, etc), samples are then
collected and sent to laboratories for confirmation tests. Confirmation requires high sensitivity and
selectivity toward drugs, as well as their metabolites, and is frequently carried out by GC/MS. Gas
chromatography (GC) is based on the separation of volatile samples by their unique affinity for the
column. Target drug compounds in the sample are identified by their retention times when samples are
passed through chromatographic columns. GC coupled with mass spectrometry (MS) is a powerful
technique because structures of unknown compounds can be identified after they have been separated
by GC. Other analytical instruments, such as HPLC, FTIR, and UV/Vis are found in forensic laboratories
as complementary techniques ( http://staff.buffalostate.edu/kimj/File/Project1.htm).
OBJECTIVE: At the end of the activity, the students are expected to appreciate the proper way of how to
perform the microscopic testing for the identification of common barbiturates by crystal formation
employing Wagenaar test.
Materials and Instrument

- Electric Microscope - gloves and masks
- Glass slides and cover slips
- Pasteur Pipettes
- Gloves and Mask
Reagents:
- Socobarbital sodium - Amybarbital
- Phenobarbital - Wagenaar Reagent
1. Place a small amount of Phenobarbital powder on a glass slide.

2. Add a drop of Wagenaar Reagent.
Preparation of Wagenaar Reagent:
- Add ethyllenediamine to a 5% Copper Sulfate until solution becomes dark violet.
3. Repeat the procedure using Amytal and Seconal.
4. Observe the crystal formation and take not of the differences.
V. QUESTIONS FOR RESEARCH
1. Tabulate the three barbiturates based on the following:

a. Other names
b. RA 9165 Classification
c. Pharmacologic classification
d. Medical use of uses
e. Route of introduction
f. Signs of benzodiazepine intoxication
2. What are the chemical tests for barbiturates?
Page 78 of 81
References:
A. Textbook
Textbooks/Reference Books (minimum of five (5) books) – must be available at the UB Library
Knight, B. (2005). Simpson’s Forensic medicine (11 th ed.). USA.: C & E Publishing Inc.
Saferstein, R. (2008). Handout in Forensic science. New Jersey: Prentice Hall.
Vidal, P. (2005). War against drug abuse. Manila: Mary Jo Publishing House, Inc.
B. Journals
Forensic Science International
Journal of Chromatography. B, Analytical technologies in the biomedical and life science (2002-present)
Journal of Chromatography. B [several earlier titles] (1977-2002)
Journal of Forensic Sciences (2006-present) [NCSU subscription coverage]
Journal of forensic Sciences backfile (1972-2005)
C. Electronic Sources
Dou, J. (2008). Forensic chemistry. Retrieved on October 2010 from

http://www.lib.ncsu.edu/courses/chem441/journals.html
Lou, S. (2009). Forensic chemistry: An introduction. Retrieved on October 2010 from

https://webauth.ncsu.edu/wrap-bin/was16.cgi?affil
Forensics in the crime scene. Retrieved on January 2009 from

http://enterprise.astm.org/SUBSCRIPTION/DIGITAL_LIBRARY/JOURNALS/FORENSIC/index.html
Page 79 of 81
Module Evaluation
The learner’s feedback is vital to us. Taking into account your assessment and impression will
help us enrich the content enhance the quality your learning engagement with us.
From this view, we would appreciate if you could spend some time completing this evaluation by
checking the column you think is appropriate and then providing a qualitative response to the questions
raised in this form.
The questionnaire is anonymous and though your participation is voluntary, your utmost
cooperation is encouraged.
Once completed the results of these questionnaires will be analyzed and an overview compiled
which will be reported to the next cohort of students in the module handbook. The overview will also be
used to inform discussion at programme team conference.
INDICATORS Strongly Moderately Slightly Disagree Strongly

Agree Agree Agree Disagree
I. The Module
a. was effectively designed
b. had clear learning

outcomes
c. was well organized
d. contained relevant
information
e. had clear images
f. had sufficient parts
g. had lessons that are

related to life experiences
II. The Assessment
a. rubrics were clear
b. instructions were
comprehensive
c. was sufficiently
challenging
d. was aligned to the

lessons
e. was done within the

prescribed time
Page 80 of 81
f. had enriched my
knowledge about the
lessons
g. were of different types
h. contained critical thinking

questions
What do you like most about the module?
What could have been improved on the module?
What other things you suggest to improve the module?
How satisfied are you with the module?
Thank you.
Page 81 of 81

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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]

II. Requirements of the Course:

Week Lecture Study Guide Laboratory Study

Rubrics for the evaluation of Illustrations and Diagrams

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

March 11, 1915 -Department of Legal Medicine was created

Definition of Forensic Chemistry:

Roles of Forensic Chemistry:

Functions of Forensic Chemistry:

Scope of Forensic Chemistry

Stages in the Practice of Forensic Chemistry

Factors Contributing to the loss of physical evidence

Techniques used in Forensic Chemistry

Principles used in Forensic Chemistry

Crime Scene Vocabulary

Crime Scene Protocol

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

MATERIALS: Illustrations, References on

1. Describe and discuss the function of each instrument in forensic toxicology.

MATERIALS: Reference books

DISCUSSION (Saferstein, 2013)

Draw/illustrate the following:

Part 1: Define the following

Study of different insects

Method of Collecting Blood

Screening Tests for Blood and Blood Stain

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

2. Indirect or Reverse Blood Typing

Confirmatory tests for Blood and Blood Stain

Blood Spatter Analysis

Person is Bleeding Profusely and struggle has taken place

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

A. PERFORMANCE OF A VENIPUNCTURE: (Calaluce,2015).

QUESTIONS FOR RESEARCH

Questions for research:

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

Questions for research

A. Preparation of Takayama Reagent

B. Procedure for Takayama Test

Questions for Research

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

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

Also known as Van Deen’s test

Part 4: Differentiate Teichman’s from Takayama test

Biological Characteristics of Semen