Forensic Chemistry and Toxicology

Mr. Clint Eleazer A. Ricarse, RMT, LPT, RCHT, MAED (CAR), MLS

(ASCPI)

1. What is Forensic Science?

The word forensic comes from the Latin word forensis: public, to the forum or public discussion;
argumentative, rhetorical, belonging to debate or discussion.

Forensic science is the application of scientific methods and processes to matters that involve crime or
the public.

2. Forensic Chemistry

A branch of chemistry which deals with the application of chemical principles in the solution of problems
that arise in connection with the administration of justice, applied in the elucidation of legal problems.

3.Scope of Forensic Chemistry

This embraces a large and diversified field not only the chemical side of criminal investigation which is
generally associated to public mind but also the analysis of any material the quality of which may give
rise to legal proceedings. It is not limited to purely chemical questions involved in legal proceedings but
also invades other branches of forensic sciences notably:

1. Legal Medicine

2. Ballistics

3. Questioned Documents

4. Dactyloscopy

5. Photograph

The responsibilities of a forensic chemist

Performs analytical examination of different controlled substances such as dangerous drugs and
explosive or explosive ingredients.
Examines trace evidences to establish if they have any connection to a crime being investigated.
Analyzes products to determine if they are fakes or if they are what they claim they are for unfair trade
competition.

Conducts researches related to cases being examined.

Prepares technical reports on laboratory findings for court presentation and testifies concerning
scientific facts..

Meets with officers, prosecutors, lawyers and court officers to discuss laboratory results. Responds to
queries of all investigating units and renders expert opinion on matters related to the application of
chemical principles for the purpose of assisting investigators develop leads in their investigation.

As a matter of social responsibility, shares his/her expertise and knowledge through lectures on Forensic
Chemistry to various schools, universities, police training centers and investigating units.

An investigation needs the help of a forensic chemist in order to solve problems of universal nature.
extremely and

Purpose of Analysis

Developing investigation Mads

Establishing aggravating or mugating circumstances in the commission of the crime

Determining the degree of physicalingeries

Distinguishing murder from homicide or suicide and majority of the cases, to establish the fact
constituting criminal violation.

GOLDEN RULES IN THE PRACTICE FORENSIC CHEMISTRY OF

The six golden rules in the practice or forensic chemistry are:

1. Go slow

2. Be thorough

3. Take notes

4. Consult others.

5. Use Imagination

6. Avoid complicated theory

GOLDEN RULES IN THE PRACTICE OF FORENSIC CHEMISTRY

Go slowly: good work cannot be hurried, therefore take all the time necessary to make the case
complete, no matter how urgent it may appear or how pressing others may be for the result; it is
generally possible to adjourn a case if work cannot be finish in time.

Be thorough; make a careful and minute examination of everything and do not be satisfied with a
qualitative analysis if a quantitative one be possible; it always pays to do too much rather than too little
and it is difficult to foresee what will or will not be required in court.

Take notes; keep a full, neat and clear record of everything seen and done.

Quiz time!

1.Considered as the Father of Toxicology

a. Mathieu Orfila

b. Sherlock Holmes

c. Sir Arthur Conan Doyle

d. None of the above

2. Francis Galton

a. Developed methodology for identification using the friction ridges on fingers

b. Developed antroротесту

c. Discovered that blood typing could be used as a means of identification

d. None of the above

3.He established the fundamental principles of document examination.

a. Albert S. Osborn

b. Calvin Goddard

c. Hans Gross

d. Edgar Hoover

4. An Austrian lawyer who coined the term 'Criminalistics'

a. Hans Gross

b. Calvin Goddard
c. Albert Osborn

d. Edgar Hoover

5. Which of the following is mismatched?

a Francis Galton Fingerprinting

b. Edmond Locard: Exchange Principle

c. Hans Gross: Criminalistics

d. None of the above

History of Forensic Science

Sherlock Holmes

Fictional character developed by author Sir

Arthur Conan Doyle.

First to apply serology, fingerprinting,firearm identification, and questioned- document examination

All of this - from fiction - before accepted by real-life criminal investigations.

Mathieu Orfila - 1813 -Father of Toxicology

Lafarge Trial (Marie) - arsenic Book Traite des poisons or Toxicologie general - first scientific study of the
detection and pathological effects of poisons - established toxicology as a distinct field of forensic
science.

Alphonse Bertillon - 1879

The father of criminal identification Developed anthropometry - using body length measurements as
unique properties

Francis Galton(1822-1911)

Fingerprinting

Developed methodology for identification using

the friction ridges on fingers

His work has been enhanced by Sir Edward Henry-father of modern fingerprinting

Henry developed a filing method for storing fingerprint patterns/records prior to computer storage
capabilities

Leone Lattes-1915

Blood testing- used technique developed by Dr. Karl Landsteiner Bloodstains-developed method to
determine blood type from dried blood Discovered that blood typing could be used as a means of
identification

Calvin Goddard -1920s

Individualization of weapons

Firing pin marks on shell casings

Refined the technique of bullet comparisons with the gun from which it was fired.

Albert S. Osborn (1858-1946)

Handwriting expert established the fundamental principles of document examination. considered the
father of the science of questioned document examination in North America

Hans Gross (1847-1915)

-First real life scientific detectivedescribed the application of scientific disciplines to the field of criminal
investigation

-Austrian lawyer

-Coined the term

Criminalistics -Wrote about forensic medicine toxicology, serology, ballistics, and anthropometry

-Suggested using mineralogists, ecologist, and botanists

J. Edgar Hoover Reorganized the Bureau of Investigations in 1924 Included fingerprint cards
Organized a national laboratory that aimed to offer forensic services of all law enforcement agencies

Edmond Locard (1877-1966)

-Locards Exchange Principle whenever two objects come in contact, material will be exchanged between
them.

-Every criminal can be tied to crime by dust particles carried from the scene

-Ex counterfeit coins metal found that matched those of the coins found on 3 suspects
clothesconfronted with evidence - they confessed

What is the importance of the data/information collected by the Forensic Scientist?

EVIDENCE

objects, documents, official statemen ts, etc. that are used to prove something is true or not true.
(Cambridge Dictionary, 2020)

It is the means, sanctioned by law, of ascertaining in a judicial proceedings the truth respecting a matter
of fact.

When a crime is committed and the assailant has not been caught, evidence becomes necessary. This is
known as the "Locard's Exchange Principle." This principle states that whenever a person enters an
environment or place of business, something is added and something is removed. In other words,
everyone leaves behind evidence or a trace of evidence.

BASIC FORMS OF EVIDENCE

Direct Evidence - that which the senses perceive. Any fact to whithta witness testifies based which a
witness testifies based on what he saw, heard, smelled, touched or tasted, is smelled, touched or tasted,
is direct evidence.

Circumstantial Evidence - a Circumstantial Evidence kind of evidence which seeks to establish a

conclusion by inferences from proved facts.

Hearsay evidence - a statement made by a witness on the authority of another and not from authority of
another and not from others.
Hearsays are generally not admissible as evidence in common law courts, but there are many exceptions
for their acceptability.

SCIENTIFIC EVIDENCE SCIENTIFIC EVIDENCE

The means sanctioned by law, of ascertaining in a judicial proceeding the truth

The means sanctioned by law, of ascertaining in a judicial pespeating a matte resploding a matter of fact
wherein scientific knowledge is necessary.

FORMS OF SCIENTIFIC EVIDENCE

Real or Authentic evidence -evidence which is addressed to the senses of the court.

Testimonial evidence -comes from people. E.g. testimony of an expert witmessimconsult

Experimental evidence

An expert witness may be required to perform

An expert witness may be required to perform certain experiments to prove a certain matter of fact.

Documentary evidence -any written evidence presented by an expert in court.

Incriminating Evidence

An evidence that may prove or disprove the involvement in a crime or fault.

Incriminate to charge with or show evidence or proof of involvement in a crime or fault (Merriam-
Webster, 2020)

Fingerprints

Since no two people have the same fingerprint, this type of evidence is one of the most reliable form of
identification. If left at the crime scene, successfully lifting and preserving this evidence will narrow
down the identity of suspects and help investigators focus on most likely persons connected to a crime.

DNA (Deoxyribonucleic Acid) Profile

Deoxyribonucleic acid (DNA) is the master molecule of every cell. The drawback with DNA evidence is
that it takes weeks or months to analyze and is a relatively expensive procedure. And because of this, it
is not used in every criminal Gane

DNA evidence was used to identify many of the 9/11 victims. When the World Trade Center was
attacked, there was no sophisticated means to identify more than 500 victims at one time. The National
Institutes of Health and other institutions used DNA profiling. In addition to receiving 20,000 pieces of
human remains from the disaster site, a database of the victims' DNA profile was used Out of the 2,792
people known to have perished, only 1,585 remains were identified by DNA extraction methods. Many
more remain unidentified to date.

Blood

DNA can be extracted from blood evidence because blood consists of cells. Blood spatters can also help
investigators piece together crime scenes and it provides theories about how a crime was committed.

Hair

Hair is another useful type of evidence that can bring police closer to a criminal. If a hair strand's root is
intact, it can be submitted for DNA testing. In addition, the color of a hair strand can also be used to rule
out suspects whose hair does not match the recovered hair sample.

Skin

This type of evidence is difficult to see at a crime scene but investigators are trained to look at the most
likely places to find this. During a crime where a victim attempts to perpetrator/criminal may find itself
under the victim's fingernails. Uke hair, skin samples can also help determine the skin. color of the
person involved in the crime and DNA can be extracted for a more accurate identification of criminals.
Skin can also help investigators understand the crime in greater detail.

Testimony from a witness who was at the scene of the crime is one of the most incriminating types of
evidence. How incriminating the testimony is depends on the credibility of the witness, which is
determined by the court. It can be the deciding factor in a case.

Witness Testimony

Written Documents

Criminals usually chronicle their plans and actions in journals, diaries, letters, and even emails. They may
confess their involvement or guilt in letters written to friends, or members of their families. When found
during searches, these can be very incriminating.

Semen

Semen can be used in several ways to verify rape accusations, as well as extract DNA to identify the
criminal. Semen can also be analyzed and tested to determine if there was more than one person
involved in a sexual crime.
Shoe Prints

Shoe prints are extremely useful in police investigations, and they can be a very incriminating type of
evidence. Police can tell a lot about a shoe print, such as the make, model and size of a shoe, as well as
the gender and approximate height of the person. Shoe prints also indicate the activity of the wearer
when the print was made and, if the impressions are visible, police may be able to trace the criminal's
moves and follow their prints to the next destination, such as a nearby home or woods.

Videotapes/ Photographs

Videotapes and photographs are both compelling and incriminating types of evidence. Whether the
videos or photos capture the crime taking place or the people present during or after the crime, it is
valuable evidence that can be used to rule out suspects and find the criminal. The identity of the
suspects in the Boston Marathon Bombing that happened last April 2013 was made possible by
collecting all footages from cellphones, CCTVs installed outside buildings proximate to the bombing as
well as aerial shots that were compared and analyzed.

Ballistics.

Ballistics is the study of firearms and ammunition. This technical form of evidence includes shell casings,
gun powder, bullets, gunshots and other firing characteristics of a weapon. Even the slightest remnants
of a gunshot can be traced to a specific firearm, where it's sold and its owner, if registered.

Drugs.

Chemical substances that could have been ingested or administered to victims can be traced in the body
of the victim through analytical laboratory tests. When the victim is deceased, tissue samples of body
organs can be subjected to laboratory tests to determine the absence or presence of drugs or chemical
substances. Whether the level of the drug indicate criminal intent, accidental administration, self-
inflicted, or abuse must be tied up to other circumstances to arrive at scientific and acceptable
conclusions.

Toxicological Reports

Toxicological reports are done by a forensic chemist based on tests. conducted on the victim
[particularly visceral organs obtained during autopsy] With these reports, investigators can determine
what type of substances were in a person hefore and at the time of the crime.

Witness

A person who has personal knowledge of something. (Merriam-Webster, 2020)

A person who testifies in a cause or before a judicial tribunal, (Merriam-Webster, 2020)

A witness in court may be an ordinary or expert witness.

DID YOU KNOW?

It is not illegal to witness a crime or to have knowledge relevant to an event and not report it. There is
no legal duty to do so. It depends on the person to report it to the police or not, most especially if
special circumstances exist. It is more out of a moral obligation. to report a crime that has been
witnessed than it is a legal obligation to do so

An Ordinary Witness

The Rule of Court requires that a person must have the following:

• must have the organ and power to perceive.

gathered other by his organ of sense can be inparted to

• does not fall in any of the exception provided for by law in not fall any of

... to be considered as ordinary witness.

An 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.

This kind of witness is called EXPERT WITNESS.

This kind of witness is called EXPERT

ORDINARY WITNESS

Can only state what his senses have perceived

may not be skilled on the line he is testifying

EXPERT WITNESS

may state what he has porcold and alsu give opinien, deduction or conclusians to bi perception

must be skilled in the art, science or trade he is testifying.

ORDINARY WITNESS

VS.
EXPERT WITNESS

Ordinary Witness

cannot testify on things or facts he has not perceived except those provided for provided by law.

Expert Witness

may testify on things which he has not seen by giving his opinions, deductions or conclusions on the
statement and conclusions on the of facts.

Values of Expert Testimony:

The degree of learning of the witness

The basis and logic of his conclusion

Other proof of the case

Four Stages of Work in the Practice of Forensic Chemistry

Collection or reception of the specimen to be examined

The actual examination

The communication or results of the examination

Court Appearance

Quiz time!

Forensic Chemistry is a branch of chemistry that deals with:

a. Application of chemical principles for the solution of a crime

b. Administration of justice

c. Convicting the guilty and exonerating the innocent

d. All of the above

A forensic chemist can testify in court as to:

a. His theory of how a crime was committed

b. The identity of the victim

c. The guilt of the suspect

d. The validity of the tests that he conducted

Result of forensic examination conducted on evidence gathered may be communicated to:

a. Person who requested for the test

b. Family of the crime victim

c. Interested military personnel

d. NBI

Which is not a responsibility of a forensic chemist?

a. Conducts chemical examination of explosives or explosive ingredients

b. Conducts DNA tests on blood or other body fluid specimen

c.Files case in court based on result of chemical examination

d.Renders expert testimony in court to back up results of tests.

A forensic chemist, when called upon to testify in court is considered an authority regarding:

a. The identity of the suspect

b. The identity of the victim

c. The guilt of the suspect

d. The validity of the tests conducted

A policeman was patioiling when ne nearu d scream, saw a man coming out of a house carrying a
bloodied knife. Upon responding, he finds a woman with a stab wound on the back apparently dead.
The policeman can testify that:

a. He knows the killer to be the man carrying the bloody knife.

b. He saw a man coming out of the house with a bloodied knife.

c. The scream he heard was that of the woman found dead.

d. The victim died of a stab wound sustained on the back.

A doctor of medicine is called upon to render testimony in court because he witnessed a crime. Because
he is a doctor, he is considered as:

a. Ordinary witness

b. Character witness

c. Expert witness

d. Eyewitness

There are golden rules in the practice of forensic chemistry. These rules are intended to ensure
thoroughness and not overlook anything that might be significant to a case. Which of the following is an
unacceptable practice?

a. Entertain even the most complicated theory

b. Have a wide and uncontrollable imagination

c. Make a record of everything seen and done

d. Good work must not be hurried.

One of the responsibilities of a Forensic Chemist is to conduct lectures in schools, police training centers
and investigation units

a. As additional source of income for Forensic Chemists

b. To share their knowledge and elevate awareness of investigators to help them perform their jobs
well.

c. A requirement of the PRC for renewal of their professional license

d. None of the above

The responsibilities of a Forensic Chemist begin with the reception of evidence from the evidence
custodian and continue up to the court if they are called to testify. Proper reception of evidence involves
the following EXCEPT:

a. Break and dispose the original seal.

b. Enter into the laboratory logbook the request.

c. Document the appearance of the sample received in its original container.

d. Carry out the requested laboratory examination.

A kind of evidence which seeks to establish a conclusion by inferences from proved facts.

a. Indirect evidence

b. Circumstantial evidence

c. Direct evidence

d. Both A and B

A witness is called upon to testify in court. As an ordinary witness, he must give his perception of:

a. What he saw

b. What he thinks

c. What he believes

d. None of the above

SAMPLING AND HANDLING TECHNIQUES

In sampling and handling of evidence, an over-arching principle is that the investigators are not a source
of contamination. Secondly, they should be knowledgeable about sampling techniques as well as
handling techniques in order to safeguard the value of the evidence to be of any use especially if the
case is litigated. Handling also includes following the chain of custody in safekeeping of evidence.
General Principles of Collecting Specimen

• Sufficiency of Samples

• Maintenance of Individuality

• Standard for comparison

• Labeling and Sealing

Sufficiency of Samples.

Most analytical tests require only a small amount of sample. But this should not mislead investigators to
be content with very minute samples during the collection process.

How much or how little will depend on the classification of the crime and the type of evidence to be
collected.

Standard for Comparison

This serves as a BLANK when the sample is analyzed. As a blank, the standard will be separately
scrutinized, analyzed and compared to the sample evidence. What will be considered significant will be
the difference between the Standard and the Sample.

Maintenance of Individuality.

When collecting samples, each evidence must be treated as a separate piece. This means that samples
must not be lumped into a common container. One piece of evidence must be packed in a separate
container and so are the other pieces of evidence.

Evidence has to be labeled properly in order to identify what it is, its source, person responsible for
collecting the same, date, time and place it was collected and if the case number is known to be
included as well. If other pertinent information is available, to include the same. Labeling will prevent
the sample from being mixed-up with similar

samples and potentially lose its value if it could not be identified later. Proper sealing of the container
will discourage and eliminate the possibility of tampering and thereby preserve the value of the
evidence.

Investigators follow their own chain of custody in handling and safekeeping of evidence. All pieces of
evidence gathered at the crime scene are turned over to the evidence custodian. Evidence that has to
be tested in the laboratory by a forensic chemist are properly turned over to the custody of the
laboratory. The forensic laboratory records all received as well as released evidence in an official
logbook of evidence. Just as investigators are not allowed to keep in their possession any evidence, the
same principle applies to forensic chemists.
Forensic chemists usually perform their analytical work in a sterile laboratory decreasing the risk of
sample contamination. In order to prevent tampering, forensic chemists must keep track of a chain of
custody for each sample. A chain of custody is a document that stays with the evidence at all times.
Among other information, the identifying tag contains signatures and identification of all the people
involved in transport, storage and analysis of the evidence.

This makes it much more difficult for intentional tampering to occur, it also acts as a detailed record of
the location of the evidence at all times for record keeping purposes. It increases the reliability of a
forensic chemist's work and increases the strength of the evidence when presented in court

Scene Investigation and Operation

Crime scene

The area or vicinity of occurrence of the crime and where physical evidence is found. It includes all areas
in which the criminal, the victim, and any eyewitnesses moved during the time of the crime and
thereafter.

The crime scene is what is subjected to a search and thorough documentation. Crime scene analysis is a
slow, methodical, systematic, and orderly process of collecting physical evidence based on proper
recognition and collection procedures. Samples collected are submitted to laboratories for exam ination
by forensic exam iners to establish or uncover significant information or clues that may provide leads to
the solution of a crime. Crime scene investigation also includes the identification and interview of
witnesses and the arrest of suspect or suspects at the crime scene.

Crime laboratories do not solve crimes. But the concerted efforts of professional and police officers who
conduct a com petent investigation will enhance chances for the successful outcome of a crim inal
investigation. The success of an investigation depends greatly upon the ability of an investigator to
recognize and preserve physical evidence. Failure to do so, no sophistical machine in the laboratory or
technicalexpertise can salvage the situation.

Crime Scene Investigation Process

1. First Investigators on the Scene

2. Crime Scene Classification

3. Documentation

4. Gathering Evidence

5. Reconstructing a Crime Scene

Processing the Crime Scene

1. Preparation for the Operations

2. Secure and Preserve the Crime Scene

3. Preliminary Survey of the Crime Scene

4. Documentation of the Crime Scene

5. Crime Scene Search

6. Collection of Evidence

7. Final Survey and Release of the Crime Scene

Documentation of the Crime Scene

a. Basic Premises

b. Approach

C. Secure and Protectd. Speak to witnesses

e. Evaluate Physical Evidence Possibilites

f. Narrative

g. Photography

h. Sketch

Crime Scene Search

a. Quadrant or Zone Search Method

b. Strip of Line Search Method

C. Spiral Search Method

d. Grid or Double Strip Search Method

e. Wheel Search Method

Spiral method (a)-may move inward or outward; best used where there are no physical barriers
(b) Strip/Line

- Line or strip method (b)- best in large, outdoor scenes

-Grid method (c)-basically a double-line search; effective, but time-consuming

- Zone method (d)-most effective in houses or buildings; teams are assigned small zones for searching

(c) Grid

(d) Zone/Quadrant

- Wheel or pie method (e)- best on small, circular crime scenes

Forensic Serology

BLOOD

The circulating tissue of the body

Complex mixture of cells, enzymes, proteins and inorganic substances.

Red fluid of the blood vessels.

Normal pH is 7.35 to 7.45

Composition of Blood

1. 45% Cells

a. Red Blood Cells (Erythrocytes)

b. White Blood Cells (Leukocytes)

C Blood Platelets (Thrombocytes)

2. 55% Plasma

a. Water (90%)

b. Solid (10%)

Introduction to Blood

White Blood Cells (LEUKOCYTES)

DNA in nucleus
Red Blood Cells (ERYTHROCYTES)

No DNA in nucleus

Blood Smear Leishman

blood platelets

Platelets (THROMBOCYTES)

erythrocytes

Forensic Analysis of Blood

1. Visual examination of evidence

Presumptive screening test (Is it blood?)

Confirmation test (Seriously, is it blood?)

Determine species origin (human blood?)

Identify the blood (whose blood is it?)

Forensic Analysis of Blood

1. Visual examination of evidence

2.Presumptive screening test (Is it blood?)

Confirmation test (Seriously, is it blood?)

Determine species origin (human blood?)

Identify the blood (whose blood is it?)

Presumptive Screening Tests

• Negative result means the questioned stain is not likely blood

Positive result means the questioned stain is likely blood

Presumptive Screening Tests

Presumptive tests produce a color reaction or release of light

Tests rely on catalytic properties of blood (hemoglobin presence)

Presumptive Screening Tests

Color Tests

• Apply chromogen (color changing chemical)

• Apply oxidizing agent (hydrogen peroxide)

• The catalyst of the reaction is hemoglobin

Rapid color change is a positive result.

This means the stain is blood.

Presumptive Screening Tests

Color Test Method

• Sample stain with clean cotton swab

• Add drop of chromogen

• Add drop of hydrogen peroxide

Presumptive Screening Tests

Color Test Method

• Sample stain with clean cotton swab

• Add drop of chromogen

• Add drop of hydrogen peroxide

Alternatively, collect stain on thread; add chromogen and hydrogen peroxide in spot plate

Presumptive Screening Tests

Color Tests

Remember, other non-blood substances might catalyze the reaction also.

Chemical Oxidants

Plant materials

False Positive - A positive result given by a substance that is not blood.

Presumptive Screening Tests

5 Types of Color Tests

Benzidine

• Phenolphthalein

• O-Tolidine

• Tetramethylbenzidine (TMB)

• Leucomalachite Green (LMG)

Presumptive Screening Tests

Benzidine

Positive result = blue color

Carcinogen (cancer causing) in 1974

No longer used

Presumptive Screening Tests

Phenolphthalein

• Positive result = pink

Some other substance produce colors other than pink (not blood) Still used today
Presumptive Screening Tests

O-Tolidine

• Positive result = blue

Similar to benzidine; still carcinogenic as it can be metabolized to benzidine No longer used; gradually
replaced by TMB

109

Tota

Presumptive Screening Tests

Tetramethylbenzidine (TMB)

• Positive result = Blue-green

Most common test for blood

1. Rub stain with moist swab

2. Add TMB

3. Add hydrogen peroxide

Look for quick blue color

Presumptive Screening Tests

Leucomalachite Green (LMG)

• Positive result = Green

Not as sensitive as TMB or specific as phe- nolphthalein

Presumptive Screening Tests

Tests Using Chemiluminescence and Fluorescence

Chemiluminescence - light is emitted as a product of the chemical reation

Fluorescence light is emitted when a substance is exposed to a shorter wavelength of light(UV light)

Presumptive Screening Tests

Chemiluminescence and Fluorescence

• More sensitive than color tests

• May damage blood stain (no blood/DNA typing)

Used to locate and define areas of blood

old blood stains

cleaned floor

Presumptive Screening Tests

Luminol

• Method similar to color test

Luminol is combined with oxidant and sprayed over area thought to contain blood

Emits a blue-white to yellow green glow

Fluorescein

• Fluorescein is combined with oxidant and sprayed over area thought to contain blood
Fluorescein when treated with a UV light

Fluorescein includes a thickener; this makes it more effective on vertical surfaces

Study showed no interference with DNA analysis

Forensic Analysis of Blood

1. Visual examination of evidence

Presumptive screening test (Is it blood?)

Confirmation test (Seriously, is it blood?)

Determine species origin (human blood?)

Identify the blood (whose blood is it?)

Confirmatory Tests

Due to the possibility of false positives with the presumptive tests, confirmatory tests are necessary

• Confirmatory tests involve making crystals that detect the presence of hemoglobin

Confirmatory Tests

Teichmann Test and Takayama Test

Small amount of blood added to microscope slide

Chemical solution is added

Slide is heated to form crystals

Crystals viewed under microscope

Confirmatory Tests

Takayama Test

Teichmann Test
Forensic Analysis of Blood

1. Visual examination of evidence

Presumptive screening test (Is it blood?)

Confirmation test (Seriously, is it blood?)

4. Determine species origin (human blood?)

Identify the blood (whose blood is it?)

Species Origin

Most methods test for serum proteins

Serum proteins are found in all animals, but are slightly different

Species ID methods based on antigen/antibody interactions

• Antigen = serum protein

Antibody produced when foreign serum protein is detected

Certain antibody will only attach to one species' serum protein

Species Origin

Antibody

Antigen

Antibody is in antiserum

Antigen (serum protein) is in blood sample

• Human antiserum will only attach to human blood sample

Rabbit antiserum will only attach to rabbit blood

sample

Dog antiserum will only attach to dog blood sample

Species Origin

Ring Precipitin Test

Blood sample (dilute) in top layer

Antiserum in heavy

Ring Precipitin Test

Blood sample (dilute) in top layer

Precipitate means blood and antiserum species match

Antiserum in heavy bottom layer

Yes line = Yes human blood

Human Antiserum

Species Origin

Ring Precipitin Test

No line = Not human blood

Human Antiserum

Forensic Analysis of Blood

1. Visual examination of evidence

Presumptive screening test (Is it blood?)

Confirmation test (Seriously, is it Deedmine species origin)

(human Identify the blood (whose blood is it?)

Forensic Analysis of Blood

1. Visual examination of evidence

Presumptive screening test (Is it blood?)

Confirmation test (Seriously, is it blood?)

Determine species origin (human blood?)

5. Identify the blood (whose blood is it?)

Testing for Seminal

Stains

Many of the cases sent to a forensic laboratory involve sexual offenses, making it necessary to examine
exhibits for the presence of seminal stains.

Characterization of Seminal Fluid

Normal male can ejaculate 2.5-6 ml of seminal fluid Each ml contains 100 million or more spermatozoa

Sperm Cell Structure

A normal sperm will have the following

characteristics:

A smooth oval head - The head of a normally formed sperm has a smooth surface and resembles the
shape of an egg

The head of the sperm measures 2.5 to 3.5 um in diameter and 4.0 to 5.5 um in length
(um=micrometers). This results in a 1.50 to 1.70 length to width ratio

a 1.50 to 1.70 length to width ratio

They have a well-developed acrosome that covers 40 to 70 percent of the oval shaped head

A slim middle section (body) that is approximately the same length as the head

A thinner tail section that is about 45 micrometer in length

https://www.microscopemaster.com/sperm-cells.html

Collection Method

Handling of articles bearing stains should be done very carefully to avoid damage to spermatozoa.
Vaginal/anal/penile swabs should be sent along with their smears on slides. Swabs should be taken on
sterile gauze / cloth and their smears prepared on sterile slides. These should be dried in air at room
temperature (37 degrees Celsius) and swabs dispatched in sterile test tube and slides in clean wrappers.

Methods Applied for Detection of Seminal Stains are Classified as:

1. Physical Examination

2. Chemical Examination

3. Microscopic Examination

Physical Examination

Include Visual Examination.

• To naked eye seminal stains generally appear translucent or opaque spots, at times with

yellowish tint and darker border depending on colour and thickness of substrata, which, if absorbent,
also acquire stiffness due to dried semen.

On good substrata seminal stains may appear to be fluorescent under ultraviolet light.

Chemical Examination

PH 74 Alkaline

The tests used to detect Seminal Stains are:

Florence Test

Barberio Test

Acid Phosphatase Test

MUP Test

LDM Isoenzyme Method

LDM Isoenzyme Method

Creatinine in Phosphokinase
Ammonium Molybdate Test (Phosphorus)

Semen Specific Glycoprotein (P30) Test

Enzyme-linked immunosorbent assay (ELISA), the SEMA® assay, for a seminal vesicle-specific antigen
(SVSA)

Florence Test

Basis: Choline is detected in this method.

Procedure: A few drops of watery solution of the stain is extracted and taken on a slide and a drop of
Florence reagent (8%) W/V solution of lodine in water containing 5% W/V of Potassium lodide) is poured
& allowed to mix slowly under a cover slip. Dark brown crystals of choline

periodide, generally needle snaped, formed with a few minutes. Non-specifics & false negative results
are common.

Barberio Test

Basis: Detection of Spermine

Procedure: A few drops of Barberio's reagent when added to spermatic fluid produces crystals of sperm
in picrate (needle shaped, rhombic & of yellow color).

For various valid reasons, like non-specificity and lack of reproducibility, the Florence and Barberio's
tests have not been accepted universally.

Yellow needle-shaped crystals

Acid Phosphatase Test

Acid Phosphatase is an enzyme secreted by the prostate gland into the seminal fluid

Can be detected when mix with an acidic solution of sodium alpha naphthylphosphate and Fast Blue dye

Bladder

Seminal Vesicles
Prostate

Urethra

Rectum

Procedure

► Moisten filter paper with water and rubbing it over the suspect area transfers any acid phos to the
filter paper

Place 1-2 drops of test solution on the filter paper

► Purple indicates acid phosphatase is present (positive reaction)

► A negative reaction produces no color change and is interpreted as an absence of semen

I don't know how that semer got in my

False results

Some vegetables (cauliflower) and fruit

(watermelon, grapes), fungus, contraceptive

creams, and vaginal secretions

These all give a positive result to the acid phosphatase test

But, none react with the speed of semen

A reaction time of less than 30 seconds is considered

a strong indication of semen

Anything longer is not considered to be semen

MUP Test

Another substance, MUP will fluoresce under UV light when it comes in contact with acid phosphate
(positive for seminal fluid)

MUP stands for 4-methyl umbelliferyl phosphate

CHOLINE AND SPERMINE TEST:

Bases: Unique combination of choline & spermine is present only in semen.

• Liquid semen & dried seminal stains can be identified by a thin layer chromatographic Technique.

• 1 ml of semen present can be detected by this method.

Microscopic Examination

Spermatozoa Test

• Semen is diluted with water and dried on filter paper

• Microscopic examination looks for spermatozoa

Sperm Issues

►Male normally releases 250-600 million sperm during ejaculation

► Chance of locating one is high, but not always true

Sperm may bind tightly to cloth

Stain must not be rubbed too hard, or it may disintegrate sperm

► Sex crimes may involve males with low sperm counts, oligospermia

Or men that have no sperm at all in their semen, aspermia

Determining time of sexual attack

Living, motile sperm survive 4-6 hrs in vaginal cavity of a living female

Non-motile sperm may be found 3-6 days after intercourse

Intact sperm (with tails) not normally found 16 hrs after sex, but have been found 72 hrs after in rare
instances

Finding seminal acid phos. decreases with time

after sex, with little chance of finding it after 48 hrs

Investigators should determine when and if voluntary

Investigators should determine when and if voluntary sex last occurred before sexual assault

PSA, or p30, will not be detected beyond 24 hrs

after sex
Rape Evidence

Presence of seminal fluid

Physical injuries such as bruising or bleeding confirms a violent sexual assault took place

Transfer of physical evidence-blood, semen, fibers, and hair-are usually present.

Collection of Rape Evidence

All outer and undergarments are collected and packaged separately in paper bags

Trace evidence is collected by standing on a clean sheet while removing clothing Bedding may be
recovered if seminal stains are present

Medical Examination of the victim

Pubic Combings

Pubic Reference Samples from victim

Vaginal swabs and smear

Rectal swabs

Medical Examination of Victim

Oral swabs

Head hairs

Blood sample

Fingernail scrapings

All clothing

Urine specimen

DID YOU KNOW?

In 1835, Henry Goddard was asked to investigate a burglary in Southampton, England. The butler
claimed to have struggled with the burglars and discharged his weapon in the process. Goddard
retrieved the bullet and by bullet comparison disproved the butler's claim. Like most gun owners in
those days, the butler made his own bullets by melting lead and pouring it into a mould. His mould had a
fault, a small pimple. Faced with the evidence, the burglar confessed to faking the burglary to gain favor
with his employer.
The Chemistry of Gunpowder

The formulation of gunpowder has one specific purpose and that is to create the most efficient and
rapid gas that would serve as propelling force that would push the bullet through the barrel of the gun
towards its intended target.

Gunpowder, in its most basic form usually has three (3) ingredients: a fuel source, an oxidizer and
stabilizer. The fuel source is charcoal and graphite. The oxygen needed for the reaction is supplied by
Potassium Nitrate (KNO;) or more commonly called salitre, a local preservative for tocino or pork; and
Sulfur as the stabilizer which speeds the rate of combustion and helps lower the temperature that the
mixture needs to ignite.

What is GSR?

When a gun is fired, there is combustion of both the primer and powder of the cartridge. Residue from
this combustion is known as gunshot residue or GSR. This residue may be deposited on a person at close
proximity to the fired gun. It may be deposited on the skin or clothing of the person who fired the gun,
or at the entrance wound of a victim, or on other target materials at the scene. There are two types of
GSR namely:

What is GSR?

There are two types of GSR namely:

1. Primer Gunshot Residue or pGSR which comprises of fused particles of the elements Lead (Pb),
Barium (Ba) and Antimony (S). PGSR is used to associate a person with a firearm.

2. Powder Gunshot Residue or GSR refers to the pattern evidence caused by un-burnt and partially burnt
gunpowder that is evident on clothing and entry wounds. This is used to that is evident on clothing and
entry wounds. This is used to determine muzzle to target distance.

Primer residues adhere to fired bullets up to a considerable distance from the muzzle some 200 meters
thus it is possible to associate a bullet with a firearm by testing for the PGSR. The presence of pGSR is
also an indicator of the proximity of the victim from the shooter.

Modern gunpowder or 'smokeless' gunpowder can contain up to 23 organic compounds but

Nitrocellulose is virtually always present along with other compounds containing nitrate or nitrogen.
Diphenylamine is a Nitrogen compound used as stabilizer in the powder and can be detected using
reagents specific for testing Nitrogen.

Collection, Preservation and Packing of Evidence

Samples must be obtained from the skin of a victim at scene at the first opportunity. Delay in obtaining
residues, movement or washing of the body prior to autopsy will considerably diminish or destroy
gunshot residues. Clothings should always be retained on the body up to autopsy. Clothings will be
examined for gunshot residues, entrance aid scene. analysis wounds of and in interpretation of the

Collection procedure:

Wear a pair of surgical gloves to ensure that you are not a transfer agent.

• Swab the back of the suspect's left and right hand using cotton balls moistened with dilute nitric acid.

Swab the back of the suspect's left and right hand using cotton balls moistened with dilute nitric acid
Perform the same on the palm areas. If a rifle or shotgun in involved, obtain also a swab of the left and
right facial cheek areas of the suspect

Put the swabs in separate plastic bags labeled hand palm cheek accordingly marked with your initials,
the date and exhibit number.

Swabs don't have to be collected if more than 72 hours has lapsed.

beled control swab. 'Get a swab of the interior of a spent casing using water instead of acid. Put the
swab

Collection procedure:

• Put clothing evidence intended for gunshot residue testing inside a plastic bag.

Mark the bag with your initials, the date and exhibit number.

• Photograph the weapon before you handle it. Handle with surgical gloves. Drop the firearm inside a
plastic bag, label accordingly and send to the laboratory. If unspent ammunition is available, send it
together with the weapon.

Do NOT remove clothing from the victim's body. Prior to examination for gunshot residue, clothing will
be subjected to photography for analysis

Do NOT remove clothing from the victim's body. Prior to examination for gunshot residue, clothing will
be subjected to photography for analysis of entrance wounds and gunshot range determination.

An alternative method is to BAG THE HANDS of the suspect by means of sandwich bags slipped over
each of the suspect's hands and secured with a rubber band.

The essential consideration is that every possible means of preventing cross examination be
implemented. Investigators should be able to answer YES to both questions below:

• Did I take all necessary steps to prevent cross examination?

• Did I wash my hands before performing the collection procedure?

Tests for Gunshot Residue

Diphenylamine Test

Diphenylamine may be used as a wet chemical test for the presence of the nitrate ion. In this test, a
solution of diphenylamine and ammonium chloride in sulfuric acid is used. In the presence of nitrates,
diphenylamine is oxidized, giving a blue coloration. This reaction has been used to test for organic
nitrates as well, and has found use in gunshot residue kits detecting nitroglycerine and nitrocellulose.

DA test is also known as the Lunge's test, Dermal Nitrate Test, Paraffin-DPA Test, and Gonzales test
[after the person who improved the test).

Sodium Rhodizonate Test

This is a test for the presence of barium or lead residue. The hands of the suspect are swabbed with
gauze moistened with dilute Hydrochloric Acid. The gauze is cut into small pieces to which sodium
rhodizonate, distilled water and a buffer solution is added. A color change to brown- pink indicates the
presence of Barium and a color change to scarlet red indicates the presence of Lead. If a mixture of both
colors is observed, this indicates that both Barium and Lead are present.

Harrison- Gilroy Test

This is also a test for the presence of Antimony on the hands. The hands of the suspect are swabbed
with gauze moistened with dilute Hydrochloric Acid. The gauze is air- dried and then treated with
Triphenylmethylarsonium lodide. A color change to orange indicates the presence of Antimony.

Modified Greiss Test

This is used to test for nitrite residues on bullet holes on surfaces such as wood, metal, furniture,
clothing, etc. A piece of photographic paper is treated with a chemical mixture of sulfanilic acid and
alpha-naphthol in methanol to make it non-reactive to light. The photographic paper is then placed face
down against a suspected bullet hole. The back of the photographic paper is steam ironed with vinegar
instead of water in the iron. Vapours from the vinegar penetrate the paper, and a reaction occurs
between any nitrite residues on the suspected bullet hole and the chemicals in the treated paper.
Reaction will be observed as red or orange specks on the paper if nitrite is present.

Factors that have bearing on the result of Nitrate tests:

Wind. It is possible that the gunpowder residue particles may have been blown on the hand directly
from the barrel of the gun being discharged by another person.
• The age and efficiency mechanism of a gun.

Older guns are less efficient but new weapons due their efficiency ensure complete combustion during
the firing. Better combustion produces lesser un-burnt residue.

Humidity of the surrounding environment (higher humidity will result to lesser combustion thus higher
yield of gunpowder nitrates);

Protective Gloves. The use of gloves to protect the hands will result to no nitrates being deposited and
will result in a negative DPA test.

Factors that have bearing on the result of Nitrate tests:

• Other Nitrogenous Materials. Handling of materials that contain Nitrogen such as fertilizers, tobacco,
urine, cigarettes, will result to a false positive DPA test.

• Time Lapse. Nitrates in the skin pores stay for up to 72 hours only which ordinary washing cannot
eliminate. Beyond 72 hours, these nitrates are already lost and DPA tests are not conclusive;

• Gun Caliber. Some guns by their design cause greater leakage of powder during the firing over other
gun types.

DID YOU KNOW?

In 1936, King George V of England was dying of terminal cancer. While the public was led to believe he
passed away peacefully, it was later revealed that his personal physician administered a dose of cocaine
and morphine in an act of mercy killing or euthanasia. Lord Dawson, the physician had carried out
euthanasia at the King's request recorded in his diary that he decided to terminate the King's life in the
late evening so that the announcement of the king's death would first appear in the morning's Times
rather than the socially inferior afternoon papers.

WHAT ARE DRUGS?

Drug is a chemical substance that brings about physical, physiological, behavioral and/or psychological
change in a person taking it. All medicines are drugs but not all drugs are of medicinal value. Medicines
are legal drugs.

These are substances that are prescribed by doctors to alleviate, cure, relieve patients of certain
diseases and symptoms. But medical drugs taken by persons to whom they are not prescribed can be
dangerous as well.

Dangerous /Prohibited Drugs are substances that can potentially damage the brain, heart and other vital
organs. Dangerous drugs act directly on the central nervous system and therefore its effects are
immediate. When taken into the body, it brings about physical, emotional or behavioral changes such
as: ability to stay awake and do continuous activity; creates a sense of power and surge of energy;
makes people paranoid or have hallucinations; creates a feeling of peace and the absence of any care.

Regulated Drugs. Drugs that have medical value but have the potential of being abused and therefore
the government regulates its use and dispensing.

MethamphetamineS

These are stimulants. Stimulants speed up the brain and give users a sense of high' and a surge of
energy. These are swallowed or snorted. However, when taken in large doses, can cause dangerously.

High body temperature, confusion, convulsions, coma and even death.

Methamphetamines are also known as: speed, uppers, meth, crystal meth, chalk, ice, glass, Christmas
tree, and crank. These are available as tablets, solution for iniection, caked or powders. gummy

Cocaine

These are also stimulants. And like methamphetamines they produce a fast, intense feeling of power
and energy. Cocaine is either inhaled or injected directly to the vein. When its effect wears off, the
feelings of power and energy are replaced by depression and paranoia and therefore would cause the
user to crave for another dose in order to feel good again. Cocaine causes the heart to beat faster and
the blood pressure to rise. As a result, if the heartbeat is dangerously fast, this will cause heart fatigue.
and eventually lead to a fatal heart attack. powder, basa, smack.

Cocaine is also called: coke, rock, snow, low, white, toot, nose candy, ash, flake,

Marijuana

This is the most widely abused drug throughout the world. It comes from the shredded leaves, stems,
seeds and flowers of the Cannabis Sativa plant locally known as Marijuana. Marijuana is smoked in the
form of a hand- rolled cigarette, or a pipe. Like cocaine, Mariuana also raises the heart rate and blood
pressure and poses the the same dangers to the heart as cocaine does.

Marijuana is also called weed, grass, pot, chronic, joint, blunt, herb, cannabis, hashish, Mary Jane.

Inhalants

Inhalants are substances that are sniffed directly from the container. They contain volatile substances
and are abused because they give the user a sense of' high. Some popular inhalants are: paint thinners,
rugby, gasoline, felt-marker fluids, hair sprays, deodorants, spray paints. Inhalants decrease oxygen to
the brain and can lead to brain damage.

Inhalants are known by more popular names of: whippets, poppers, snappers, rush, bolt and bullet.

Barbiturates

These are classified as Central Nervous System (CNS) depressants. They are used to reduce restlessness,
emotional tension, as an anticonvulsant and anesthetic.

Heroin

Heroin is derived from morphine alkaloid that is found in opium. When taken it produces euphoria,

analgesia or relief from pain and anxiety.

It is usually injected but it can also be vaporized, sniffed, taken orally or as a suppository.

Heroin abuse will result to slow, shallow breathing, drop of blood pressure, muscle spasms, convulsions,
coma and even death.

LSD

LSD stands for lysergic acid diethylamine. It is a hallucinogenic drug. It is sold as "acid" in the form of
tablets, capsules, and sometimes in liquid form.

It is also reterred to as Lucy in the sky with diamonds, and lightning.

Amphetamines

Amphetamines are psychostimulant drugs the most famous of which is Ecstasy. When taken it produces
euphoria, alertness, increased libido, concentration energy, self-esteem, self- confidence, grandiosity. It
is used as a performance enhancer and a cognitive enhancer. College and high school students use it as
a study and testtaking aid because it increases energy levels, and motivation. which allows students to
study over longer periods than usual. It has such side negative effects as irritability, aggression,
paranoia, insomnia, hypertension. Ecstasy is also referred to as X, Love Drug, Libido, LBD. When abused
may result to coma and death.
FORENSIC TOXICOLOGY

ESSENTIAL QUESTIONS

How can you distinguish among the types of toxins and their effects on the body?

How are different forensic techniques used to isolate and identify toxins in the body?

What factors influence the effects of alcohol on the body and why?

Forensic toxicology is...

• the use of detecting and identifying the presence of drugs and poisons in body fluids, tissues, and
organs to aid medical or legal investigation.

• The primary concern for forensic toxicology is not the legal outcome of the toxicological investigation
or the technology utilized, but rather the obtaining and interpreting of the results.

History

Mathieu Orfila, chairman of the legal department at La Sorbonne in France, published a book in 1814.
medicine

• This was the first attempt to systematically study and classify poisons.

Orfila was the first to articulate the fact that poisons must be absorbed, or enter the blood, to manifest
their effects.

In the U.S., forensic toxicology did not develop until the early 20th Dr. Alexander Gettler (lead FT in NYC
medical examiner lab) is considered this. country's first forensic toxicologist.

century in New York

toxicology

FORM

The form of a substance may have a profound impact on its toxicity especially for metallic elements, also
termed heavy metals. For example, the toxicity of mercury vapor differs greatly from methyl mercury.
Another example is chromium. Cr³+ is relatively nontoxic whereas Cr6+ causes skin or nasal corrosion
and lung cancer.

The form of the substance

Insoluble vs. soluble

The innate chemical activity of substances also varies greatly. Some can quickly damage cells causing
immediate cell death. Others slowly interfere only with a cell's function.

DOSAGE

The dosage is the most important and critical factor in determining if a substance will bean acute or a
chronic toxicant. Virtually all chemicals can be acute toxicants if sufficiently. large doses are
administered. Often the toxic mechanisms and target organs are different for acuteand chronic toxicity.

ROUTE OF EXPOSURE

The way an individual comes in contact with a toxic substance, or exposure route, is important in
determining toxicity. Some chemicals may be highly toxic by one route but not by others. Two major
reasons are differences in absorption and distribution within the body.

How it enters the body

Inhalation vs. injection vs. ingestion

ABSORPTION

The ability to be absorbed is essential to systemic toxicity. Some chemicals are readilyabsorbed and
others are poorly absorbed. For example, nearly all alcohols are readily

absorbed when ingested, whereas there is virtually no absorption for most polymers. The rates and
extent of absorption may vary greatly depending on the form of a chemical and the route of exposure.
to it.

Roles of the Toxicologist

• Must identify one of thousands of drugs and poisons

• Must find nanogram to microgram quantities dissipated throughout the entire body

• Not always looking for exact chemicals, but metabolites of desired chemicals (ex. heroin morphine
within seconds)

Toxicology Procedures

• Presumptive/Screening

⚫ quick test to narrow down possibilities spot/color tests

• Confirmation

determines exact identity thin-layer/gas chromatography, IR spectroscopy, mass spectrometry

Presumptive/Screening

• Marquis Test:

Turns purple in the presence of opiates

Turns orange-brown in presence of amphetamines

• Scott Test:

Turns blue in the presence of cocaine

Duquenois-Levine:

• Turns purple in the presence of tetrahydrocannabinal

Marquis Test

Shank & W

Scott Test

Scott Cocaine Test (Simplified) POSITIVE RESULT

Duquenois- Levine

Duquenois

Marijuana Test

POSITIVE RESULT - Blue