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Toxicology
Toxicology
Formed from the Greek words
toxicos and logos, toxicology is
the study of the symptoms,
mechanisms, treatments and
detection of poisoning
Vocabulary
Forensic toxicology centers on
the determination of toxic
substances in human tissues,
organs and body fluids such as
urine and blood, and the subsequent
determination of the role any toxic
agents may have contributed to or
caused death.
3
Toxicology
Toxin is any material that is considered
to have a life threatening effect on a
living organism.
Poison is a subgroup of toxins
- comes in many forms
Gaseous
Animals
Liquid
Minerals
Solid
Vegetables
Toxicology
Types of toxicologists
Descriptive Toxicologist: Performs
toxicity test to evaluate the risk that
exposure pose to humans
Mechanistic Toxicologist: attempts to
determine how substances exert deleterious
effects on living organisms.
Regulatory Toxicology: Determines
whether or not a substance has low enough
risk to justify making it available to the
public. Forensic Toxicology is a subspecialty
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Provide Answers to ?
Forensic Toxicologists:
Provide answers to questions that may
be asked during the investigation or at
a subsequent legal hearing.
Was a poison involved in the case?
If so, what was the nature of the poison
how was it administered
was it at a potentially lethal concentration?
6
Forensic Toxicology
What do they Investigate?
Accidental Poisoning
Drug Abuse/Overdose
Suicidal/Homicidal Poisoning
*75% of the evidence
evaluated in the crime
lab is drug related!
7
Vocabulary
2 classes of toxicity: acute and chronic.
Acute toxicity refers to effects that occur
shortly after a single exposure or small
number of closely spaced exposures.
Chronic toxicity refers to delayed effects
that occur after long term repeated
exposures.
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Aniline(hypnotics,
nitrobenzene)
Atropine(Belladonna),
Scopolamine
Odor of disinfectant
CO
HCN
Food poisoning
Metallic compounds
Nicotine
Convulsions
Opiates
Odor of garlic
NaF
Convulsion
Strychnine
11
Poisonings
Children under 6 account for most of the
reported poisonings/year
About 700 deaths by poisoning are
reported each year
Approximately 2 million cases are
voluntarily reported to poison control
centers each year
However adults account for the majority
of deaths by poisoning most of which is
intentional rather than accidental
12
Toxins/Poisons
Most Reported
1 - Household cleaning supplies
2 - Analgesics (aspirin, acetaminophen)
3 - Cosmetics
4 - Cough & cold remedies
5 - Plant scrapes & insect bites
6 - Pesticides
7 - Topical creams & lotions
8 - Hydrocarbons i.e. gasoline, kerosene
9 - Anti-micro bacterial soaps
10 - Sedatives/hypnotics/anti-psychotics
11 - Food poisoning
12 - Alcohol
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Toxins/Poisons
14
Pharmacokinetics &
Pharmacodynamics
Pharmacokinetics is the relationship
between drug conc. In the body & time
after administration of the drug. Form,
frequency & route all have an effect on
this
Pharmacodynamics is the study of the
biochemical & physiological effects of a
drug and its mechanism of action.
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Solubility
Gastric emptying
Metabolic Rate
16
Structure of GI Tract
The GI tract has three major
regions: the stomach, small
intestine and the large
intestine.
Stomach the mucosa contains many
folds that increase the surface area
available for drug absorption. The
high blood supply and the fact that a
drug can potentially reside in the
stomach for several hours can
provide optimum conditions for the
absorption of acidic drugs.
17
Structure of GI Tract
Small Intestine - the most important GI site
for drug absorption. The intestinal mucosa
provides an extremely large surface area; villi
and microvilli aids drug absorption (about 30
long in an adult)
Large Intestine - like the stomach, lacks the
villi and microvilli of the small intestine; serves
as a site for the absorption of drug that has
not been completely absorbed in the small
intestine. (about 6 long in an adult)
18
Mechanisms of Drug
Transport Across the GI
Barrier
The majority of drugs cross the membrane by passive
diffusion.
Physiological Factors
Influencing Drug Absorption
Surface area of the absorption site
pH of the GI fluids
Gastric emptying rate: can be stimulated by
hunger, anxiety, body position and liquid
consumption; Fatty foods, high bulk diet,
depression and various drugs and alcohol retard it.
Intestinal motility
Drug stability in the GI system
Dietary components
Disease states
20
Drug Metabolism
Metabolism is the fundamental mechanism of
drug elimination. Metabolism aids drug
excretion processes and may affect the
pharmacological response of a drug by
altering its potency and/or duration of action
Metabolism can occur in the plasma as well
as organs other than the liver such as the GI
tract, kidneys and lungs.
21
Factors Affecting
Metabolism
Age
Young children and elderly people usually have
a lower metabolic capacity
Disease
Diseases can affect all of the ADME processes.
Liver and kidney diseases probably have the
greatest effect on drug concentrations by
affecting metabolism and excretion.
22
Factors Affecting
Metabolism
Weight
Genetic Factors
Gender and race differences in enzyme activity can affect
drug metabolism
Diet
Diet may influence the metabolism of some drugs.
Exposure to other drugs (particularly alcohol) markedly
affects the metabolism of certain drugs.
23
Excretion of Drugs
The excretion of drugs and metabolites
terminates their activity and presence in the
body. Elimination can occur by various routes.
The kidney plays a major role with the
excretion of drugs and/or their metabolites into
the urine. Drugs may also be excreted in the
feces, bile, lungs sweat, saliva and breast milk.
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What is a drug?
A drug can be defined as a natural or synthetic
substance that is used to produce physiological or
psychological effects in humans or other higher order
animals.
Narcotic drugs are analgesics, meaning they relieve
pain by a depressing action on the central nervous
system. This effects functions such as blood
pressure, pulse rate and breathing rate.
The regular use of a narcotic drug will invariably lead
to physical dependence.
The most common source for these narcotic drugs is
opium, extracted from poppies.
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Illicit Drugs
NIDA 9
Methamphetamine
Cocaine
Opiate
Benzodiazepam
Amphetamine
THC
Methadone
Barbiturates
PCP
CNS Depressants:
Narcotics (opiates)
Sedatives, hypnotics
(PCP, Quaalude)
Tranquilizers (Valium)
CNS Stimulants
Cocaine
Amphetamines
Hallucinogens
Mescaline, psilocybin,
LSD, PCP
THC
Steroids
Inhalants
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Signs of an Overdose
Respiratory depression
cold clammy skin
Confusion
Convulsions
Severe drowsiness
Constricted pupils
28
Hallucinogens
A class of drugs is hallucinogens; marijuana is
the most well-known member of this class.
Hallucinogens cause marked changes in
normal thought processes, perceptions, and
moods.
Marijuana is the most controversial drug in this
class because its long-term effects on health
are still largely unknown.
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Hallucinogins
Cannabinoids Marijuana
The chemical substance largely responsible for the hallucinogenic
properties of marijuana is known as tetrahydrocannabinol, or THC.
The THC content of Cannabis varies in different parts of the plant,
generally decreasing in the following sequence: resin, flowers,
leaves, with little THC in the stem, roots, or seeds.
The THC-rich resin is known as hashish.
Marijuana does not cause physical dependency
the risk of harm is in heavy, long-term use.
31
Hallucinogens
Drug/Commercial & Street Names
Administered/Detection in Urine*
Hashish
Boom, chronic, gangster,
hash, hash oil, hemp
Swallowed, Smoked
14 days - 11weeks
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Other Hallucinogens
Other hallucinogens include LSD, mescaline, PCP, psilocybin, & MDMA (Ecstasy).
LSD is synthesized from lysergic acid, & can cause hallucinations that can last for
12 hours.
Phencyclidine, or PCP, is often synthesized in clandestine laboratories & is often
smoked, ingested, or sniffed.
PCP is often mixed with other drugs, such as LSD, or amphetamine, & is sold as a
powder (angle dust), capsule, or tablet.
Oral intake of PCP first leads to feelings of strength and invulnerability, which may
turn to depression, tendencies toward violence, & suicide.
33
Opiates
Morphine is readily extracted from opium and is
used to synthesize heroin.
Addicts frequently dissolve heroin in water by
heating it in a spoon, and then inject in the skin.
Heroin produces a high that is accompanied
by drowsiness and a sense of well-being that
generally last for three to four hours.
Codeine is also present in opium, but it is
usually prepared synthetically from morphine.
34
Opiates
OxyContin, with the active ingredient oxycodone, is
not derived from opium or morphine, but does have
the same physiological effects on the body as do
opium narcotics.
OxyContin is prescribed to a million patients for
treatment of chronic pain.
Methadone is another well-known synthetic opiate.
Methadone, which is pharmacologically related to
heroin, appears to eliminate the addicts desire for
heroin while producing minimal side effects.
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Depressants
Depressants are another class of drugs.
Depressants are substances used to
depress the functions of the central nervous
system.
Depressants calm irritability and anxiety and
may induce sleep.
These include alcohol (ethanol),
barbiturates, tranquilizers, and various
substances that can be sniffed, such as
airplane glue, model cement, or aerosol gas
propellants such as freon.
37
Depressants
Alcohol (ethyl alcohol) enters the bodys bloodstream
and quickly travels to the brain, where it acts to suppress
the brains control of thought processes and muscle
coordination.
Barbiturates, or downers, are normally taken orally and
create a feeling of well-being, relax the body, and
produce sleep.
Tranquilizers, unlike barbiturates, produce a relaxing
tranquility without impairment of high-thinking faculties or
inducing sleep.
Sniffing has immediate effects such as exhilaration, but
impairs judgment and may cause liver, heart, and brain
damage, or even death.
38
Benzodiazepine
Anesthetics/PCP
Ativan, Halcion, Librium, Phencyclidine: angel Valium, Xanax: candy, dust, downers,
boat, hog, love boat,
sleeping pills, peace tanks pill
Injected, swallowed
Injected, swallowed,
smoked
1-6 weeks
7-14 days
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Heroin Usage
In 1974 estimated users were 246,000.
Between the years 1988-1994 there were an
estimate of 28,000-80,000 new users
Between 1995 and 2001 the number of new
users was greater than 100,000
Today there has been a reported 3.7 million
people in the US alone who reported using
heroin at least once in their lifetime.
43
44
Oxycodone
In 1993 3.5 tons of Oxycodone were
manufactured for sale in the United
States
2003 41 tons were manufactured
The tablet is either taken orally or
crushed and sniffed or dissolved in water
and injected
This drug is often stolen and
prescriptions are often forged
45
Hydrocodone
It is used as a cough suppressant and
analgesic
DEA has this drug listed as the most
frequently encountered opiate
pharmaceutical that is submitted for drug
evidence to federal, state and local forensic
laboratories.
Abusers obtain the is drug by theft, doctor
shopping, fraudulent prescriptions, and fake
call in prescriptions
46
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Dextropropoxyphene
Related to Morphine
First marketed in 1957
Used to help moderate to mild pain
There is 150 tons produced in the United
States annually
25 million prescriptions have been written.
This drug is among the top 10 drugs reported
by the medical examiner in drug abuse deaths.
48
Stimulants
The drug classification of stimulants includes
amphetamines, sometimes known as uppers or
speed, and cocaine, which in its free-base form
is known as crack.
Stimulants are substances taken to increase
alertness or activity, followed by a decrease in
fatigue and a loss of appetite.
50
Stimulants
Amphetamine and methamphetamine, often injected
intravenously, causes an initial rush, followed by an
intense feeling of pleasure.
Followed by a period of exhaustion and a prolonged period of
depression.
52
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Toxicology of Alcohol
The analysis of alcohol exemplifies the primary
objective of forensic toxicologythe detection
and isolation of drugs in the body for the purpose
of determining their influence on human
behavior.
Alcohol, or ethyl alcohol, is a colorless liquid
normally diluted with water and consumed as a
beverage.
Like any depressant, alcohol principally effects
the central nervous system, particularly the brain.
59
Alcohol Levels
Alcohol appears in the blood within minutes
after it has been taken by mouth & slowly
increases in concentration while it is being
absorbed from the stomach & the small intestine
into the bloodstream.
When all alcohol has been absorbed, a max
alcohol level is reached in the blood; and the
post-absorption period begins.
The alcohol concentration slowly decreases
until a zero level is again reached.
60
Alcohol Levels
Factors affecting outcome on individual consumption:
1. Time taken to consume the drink
2. Alcohol content
3. Amount consumed
4. Food presence in the stomach
The extent to which an individual may be under the
influence of alcohol is usually determined by either
measuring the quantity of alcohol present in the
blood system or by measuring the alcohol content
in the breath.
61
Alcohol Levels
Elimination of alcohol throughout the body is
accomplished through oxidation & excretion.
Oxidation takes place almost entirely in the
liver, while alcohol is excreted unchanged in
the breath, urine, and perspiration.
Experimental evidence has verified that the
amount of alcohol exhaled in the breath is in
direct proportion to the blood concentration.
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Field Testing
Law enforcement officers typically use field
sobriety tests to estimate a motorists degree
of physical impairment by alcohol and
whether or not an evidential test for alcohol
is justified.
The horizontal gaze nystagmus test, walk
and turn, and the one-leg stand are all
considered reliable and effective
psychophysical tests.
A portable, handheld, roadside breath tester
may be used to determine a preliminary
breath-alcohol content.
69
Anabolic Steroids
These are synthetic compounds that
are chemically related to the male sex
hormone testosterone.
Anabolic steroids are often abused by
individuals who are interested in
accelerating muscle growth.
Side effects include unpredictable
effects on mood and personality,
depression, diminished sex drive,
halting bone growth, and liver cancer.
70
Schedules of Classification
Schedule I drugs have
a high potential for
abuse and have no
currently accepted
medical use such as
heroin, marijuana,
methaqualone and
LSD.
72
Schedules of Classification
Schedule II drugs have a high potential
for abuse and have medical use with
severe restrictions such as cocaine, PCP,
and most amphetamine and barbiturate
prescriptions.
73
Schedules of Classification
Schedule III drugs have less potential for
abuse and a currently accepted medical
use such as all barbiturate prescriptions
not covered under Schedule II, codeine,
and anabolic steroids.
74
Schedule of Classification
Schedule IV drugs have a low potential for
abuse and have a current medical use such
as darvon, phenobarbital, and some
tranquilizers such as diazepam (valium) and
chlordiazepoxide (librium).
Schedule V drugs must show low abuse
potential and have medical use such as
opiate drug mixtures that contain
nonnarcotic medicinal ingredients.
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Drug Identification
The challenge or difficulty of forensic drug
identification comes in selecting analytical
procedures that will ensure a specific
identification of a drug.
This plan, or scheme of analysis, is
divided into two phases.
Screening test that is nonspecific and
preliminary in nature to reduce the
possibilities to a manageable number.
Confirmation test that is a single test
that specifically identifies a substance.
79
Screening Tests
A screening test is normally employed to
provide the analyst with quick insight into
the likelihood that a specimen contains a
drug substance.
Positive results arising from a screening
test are considered to be tentative at best
and must be verified with a confirmation
test.
The most widely used screening tests are
thin-layer chromatography, gas
chromatography, and immunoassay.
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Confirmatory Tests
Different testing
panels may have
more then one
combination of tests.
Sodium
Nitroprusside
84
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Analytical
Instrumentation
Specific/ Non-specific:
Stimulant vs. Depressant
Exact ID of the drug
Destructive/Non-destructive:
Sample consumed, further testing
Instrumentation: GC/MS
FTIR
Chromatography
Chromatography is a means of separating and
tentatively identifying the components of a mixture.
The theory of chromatography is based on the
observation that chemical substances have a
tendency to partially escape into the surrounding
environment when dissolved in a liquid or when
absorbed on a solid surface.
Those materials that have a preference for the
moving phase will slowly pull ahead and separate
from those substances that prefer to remain in the
stationary phase.
87
TLC
TLC uses a solid stationary phase usually coated onto a
glass plate and a mobile liquid phase to separate the
components of the mixture.
The liquid will slowly rise up the plate by capillary action
causing the sample to become distributed between the
stationary phase and the moving liquid phase.
Because most compounds are colorless, the materials
must be visualized by placing the plates under ultraviolet
light or spraying the plate with a chemical reagent.
The distance a spot travels up a thin-layer plate can be
assigned a numerical value known as the R f value.
88
GC/MS
Destructive, Specific
Chromatography: selective separation
of components of a sample
GC separates
MS identifies and
quantitates
*Forensic Labs use
GC methods to test
for blood alcohols
GC/MS
Chromatography: selective
separation of components of a
sample
90
Gas Chromatography
In GC: 2 phases
The moving phase.
The stationary phase
After a mixture has traversed the length of the column,
it will emerge separated into its components.
The written record of this separation is called a
chromatogram.
The time required for a component to go through a
GC column is known as retention time.
91
Gas Chromatography
Sample diluted in solvent
Injected into capillary column
Heat vaporizes sample, travels through
column
Detector records time it takes to move
through column (retention time)
Data (chromatogram) compared to
known substances
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Gas Chromatography
Chromatograms
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Theory of Light
Light is described as a continuous wave.
When white light passes though a prism, it is dispersed into a
continuous spectrum of colors.
Waves are described in terms such as:
Wavelength, the distance between two successive crests (or
one trough to the next trough).
Frequency, the number of crests (or troughs) passing any
one given point per unit of time.
The electromagnetic spectrum is the entire range of radiation
energy from the most energetic cosmic rays to the least
energetic radio waves.
Visible light is only a small part of the electromagnetic
spectrum.
95
Spectrophotometry
Just as a substance can absorb visible light to
produce color, many of the invisible radiations of
the electromagnetic spectrum are likewise
absorbed.
Spectrophotometry, an important analytical tool,
measures the quantity of radiation that a
particular material absorbs as a function of
wavelength and frequency.
96
UVand IR
Spectrophotometry
Mass Spectrometry
In the mass spectrometer, a beam of highenergy electrons collide with a material,
producing positively charged ions.
These positive ions almost instantaneously
decompose into numerous fragments, which
are separated according to their masses.
The unique feature of mass spectrometry is
that under carefully controlled conditions, no
two substances produce the same
fragmentation pattern.
98
GC and Mass
A direct connection between the GC column
and the mass spectrometer allows each
component to flow into the mass spectrometer
as it emerges from the GC.
The separation of a mixtures components is
first accomplished by the GC.
Then, fragmentation of each component by
high-energy electrons in the mass
spectrometer, will produce a distinct pattern,
somewhat like a fingerprint, of the substance
being examined.
99
FTIR
Non-destructive, specific
Visible light is made of electromagnetic
waves, seen as colorsIR is in the
spectrum, we just cant see
Works by measuring degree of
absorption of IR waves of certain
frequencies
Works only with pure samples
100
FTIR - Printouts
Cocaine
Heroin
101
Toxicity
Water can cause fatal electrolyte
imbalances
Arsenic and cyanide are harmless in
small doses
103
Drug Identification
The challenge or difficulty of forensic drug
identification comes in selecting analytical
procedures that will ensure a specific
identification of a drug.
This plan, or scheme of analysis, is divided
into two phases.
Screening test that is nonspecific and preliminary
in nature to reduce the possibilities to a
manageable number.
Confirmation test that is a single test that
specifically identifies a substance.
107
Preliminary Analysis
The unknown substance may be any one of a thousand
or more commonly encountered drugs, the analyst
employs screening tests to reduce these possibilities to
a small and manageable number.
This objective is often accomplished by subjecting the
material to a series of color tests that will produce
characteristic colors for commonly encountered illicit
drugs.
Microcrystalline tests can also be used to identify
specific drug substances by studying the size and shape
of crystals formed
108
Conformational
Determination
Nondrug Poisons
Heavy metals such as arsenic, bismuth, antimony,
mercury, and thallium are only occasionally encountered
because severe environmental protection regulations
restrict their availability to the general public.
Carbon monoxide is one of the most common poisons
encountered in a forensic laboratory.
To measure the concentration of carbon monoxide in the
blood spectrophotometric methods determine the
amount of carboxyhemoglobin relative to oxyhemoglobin
or total hemoglobin; or a volume of blood can be treated
with a reagent to liberate the carbon monoxide, which is
then measured by gas chromatography.
113
Significance of Findings
Once a drug is found and identified, the
toxicologist determines its influence on the
behavior of the individual.
For many drugs, blood concentration levels are
readily known and are used to estimate the
pharmacological effects of the drug on the
individual.
When dealing with a living person, the
toxicologist has the added benefit of knowing
what a police officer may have observed about
an individuals behavior and motor skills.
114
ELISA Template
Confirmatory testing
115
HairMaggots
Bacteria
Anthrax
fluids
Other
body
117
Results:
A clear well indicates the presence
of the drug (The drug bound to the plate,
prevented conjugate from binding so substrate
couldnt bind and produce a color change)
119
George
Trepal