Principles of Toxicology:

The Study of Poisons

Department of
Cellular Biochemistry and
Toxicology

Toxicology

Definitions
Toxicological studies
Dose-response correlations
Threshold limit values
Examples

Toxicology

Exposure + Hazard = Risk

All substances can be a poison
Dose determines the response
Pathway, Duration of Frequency of Exposure and Chemical
determine Dose
Absorption, Distribution, Metabolism & Excretion
The extent of the effect is dependent upon the concentration
of the active compound at its site of action over time
Bioactivation: compounds to reactive metabolites
Individual variation of the organism will affect ADME

The study of the adverse effects

of a toxicant on living organisms
Adverse effects
any change from an organisms normal state
dependent upon the concentration of active compound at
the target site for a sufficient time.

Toxicant (Poison)
any agent capable of producing a deleterious response in a
biological system

Living organism
a sac of water with target sites, storage depots and enzymes

What is a Poison?
All substances are poisons;
there is none that is not a poison.
The right dose
differentiates a poison and a remedy.
Paracelsus (1493-1541)

Environmental Health Paradigm

Exposure Assessment
Emission Sources

Effects Assessment
Internal Dose

Health Effects

Environmental
Concentrations

Human Exposure

Hazardous
Denotes the probability of injury or illness
from contact or use
Industrial Hazards

Toxicity
Explosivity
Ignitability
Reactivity

Toxic Substance
Capacity of a substance to produce injury or
illness
Acute Effects
Short term, appear shortly after exposure. Can
be from single exposure

Chronic Effects
There is a latency, long period of time before
you see effect

Three Types of Toxic Hazardous

Materials
Chemical Agents (poisons)
Physical Agents (dusts, fibers, heat, noise,
corrosive)
Biological Agents (pathogens)

Definitions
Toxicology is the quantitative and
qualitative study of the adverse effects of
toxicants on biological organisms
Toxicant is a chemical or physical agent
that produces adverse effects on biological
organisms.

So Toxicology is the study of:

How toxicants enter the organism
How toxicants effect the organism
How toxicants are eliminated from (leave) the organism
All substances are toxic if taken in the wrong quantities

How toxicants enter organism

Inhalation (mouth or nose to lungs) then into
blood(+*)
Ingestion (mouth to stomach) then into
blood(+)
Injection (cuts, punctures in skin) into blood
Dermal absorption (through skin) into
blood(+*)
+ Involve membrane transport
* Greatest threats in industry

Effects of Toxicants
Irreversible Effects
Carcinogen - causes cancer
Mutagen - causes chromosome damage
Reproductive hazard - damage to
reproductive system
Teratogen - causes birth defects

Effects of Toxicants
May or may not be reversible
Dermatotoxic affects skin
Hemotoxic affects blood
Hepatotoxic affects liver
Nephrotoxic affects kidneys
Neurotoxic affects nervous system
Pulmonotoxic affects lungs

Definitions
Pharmacokinetics the absorption,
distribution, metabolism and excretion of
chemicals through the (human) system.
Bioaccumulation things such as lead,
mercury, PCBs, carbon tetrachloride that build
up in organs and have low excretion rate. Low
exposure over a long time leads to response

Elimination of toxins
Excretion through kidneys, liver and lungs
Detoxification is the biotransformation of
chemicals into something less harmful
Storage in fatty tissue

Toxicological Studies
Baseline study with no toxicant
Toxicology study to quantify response to
toxicants in specified physical state

Difficulties in Toxicological
studies
Baseline study required (control group)
Response not necessarily numerical
Specificity of individual response

Allergy or immunity
Statistical study required
Organism specific response, not applicable to humans
Dosage response
Response time, latency, acute versus chronic
Difficulty in measuring intended variable (lead in liver measured
by lead in blood)

Difficulties in Toxicological
Studies
Major Problem
No ethical way to get human volunteers, hence
need to use model systems of rats, cats, dogs,
rabbits, etc.

Hinders production of a new chemical,

almost as stringent as a new drug
Currently averages 17 years

Dose
The amount of chemical entering the body
This is usually given as
mg of chemical/kg of body weight = mg/kg
The dose is dependent upon
* The environmental concentration
* The properties of the toxicant
* The frequency of exposure
* The length of exposure
* The exposure pathway

What is a Response?
The degree and spectra of responses depend
upon the dose and the organism--describe
exposure conditions with description of dose
Change from normal state
could be on the molecular, cellular, organ, or
organism level--the symptoms

Local vs. Systemic

Reversible vs. Irreversible
Immediate vs. Delayed
Graded vs. Quantal
degrees of the same damage vs. all or none

Dose versus Response

Run test on large
population
Given same dose (usually
in dose/body mass)
Determine the number
or fraction of individuals
that have a response

Dose versus Response (cont)

Repeat tests using different
doses
Find average response to
each dose
Plot Response versus
logarithm of dose
Forms Sigmoid shaped
curve

Dose-Response Relationship:
As the dose of a toxicant increases,
4
so does the response.
RESPONSE
0-1 NOAEL
2-3 Linear Range
4 Maximum Response

DOSE
DOSE DETERMINES THE BIOLOGICAL RESPONSE

Dose Limit Values

EDf Effective dose for f
percent of population.
Reversible response
TDf Toxic dose for f
percent of population.
Undesirable response that
is irreversible
LDf Lethal dose for f
percent of population.

LD50
Quantal responses can be treated as gradient when data
from a population is used.
The cumulative proportion of the population responding
to a certain dose is plotted per dose--10-30 fold variation
w/in a population
If Mortality is the response, the dose that is lethal to 50%
of the population LD50 can be generated from the curve
Different toxicants can be compared--lowest dose is
most potent

Definitions
Therapeutic Margin
TM = LD50% - ED50%

Margin of Safety
MOS = LD5% - ED95%

Safety Index
SI = LD5%/ED95%

Therapeutic Index
TI = LD50%/ED50%

LD50 Comparison
Chemical
Ethyl Alcohol
Sodium Chloride
Ferrous Sulfate
Morphine Sulfate
Strychnine Sulfate
Nicotine
Black Widow
Curare
Rattle Snake
Dioxin (TCDD)
Botulinum toxin

LD50 (mg/kg)
10,000
4,000
1,500
900
150
1
0.55
0.50
0.24
0.001
0.0001

Exposure: Pathways
Routes and Sites of Exposure

Ingestion (Gastrointestinal Tract)

Inhalation (Lungs)
Dermal/Topical (Skin)
Injection
intravenous, intramuscular, intraperitoneal

Typical Effectiveness of Route of Exposure

iv > inhale > ip > im > ingest > topical

Exposure: Duration
Acute
< 24hr
usually 1 exposure
Subacute 1 month repeated doses
Subchronic
1-3mo
repeated doses
Chronic > 3mo
repeated doses
Over time, the amount of chemical in the body can
build up, it can redistribute, or it can overwhelm
repair and removal mechanisms

ADME:
Absorption, Distribution,
Metabolism, and Excretion
Once a living organism has been exposed to a
toxicant, the compound must get into the body
and to its target site in an active form in order to
cause an adverse effect.
The body has defenses:
Membrane barriers
passive and facilitated diffusion, active transport

Biotransformation enzymes, antioxidants

Elimination mechanisms

Absorption:
ability of a chemical to enter the blood
(blood is in equilibrium with tissues)
Inhalation--readily absorb gases into the blood stream via
the alveoli. (Large alveolar surface, high blood flow, and
proximity of blood to alveolar air)

Ingestion--absorption through GI tract stomach (acids),

small intestine (long contact time, large surface area--villi;
bases and transporters for others)
1st Pass Effect (liver can modify)

Dermal--absorption through epidermis (stratum corneum),

then dermis; site and condition of skin

Distribution:
the process in which a chemical agent
translocates throughout the body
Blood carries the agent to and from its site of
action,
storage
depots,
organs
of
transformation, and organs of elimination
Rate of distribution (rapid) dependent upon
blood flow
characteristics of toxicant (affinity for the tissue,
and the partition coefficient)

Distribution may change over time

Distribution:
Storage and Binding
Storage in Adipose tissue--Very lipophylic
compounds (DDT) will store in fat. Rapid
mobilization of the fat (starvation) can rapidly
increase blood concentration
Storage in Bone--Chemicals analogous to
Calcium--Fluoride, Lead, Strontium
Binding to Plasma proteins--can displace
endogenous compounds. Only free is available
for adverse effects or excretion

Target Organs: adverse effect is dependent

upon the concentration of active compound at the
target site for enough time
Not all organs are affected equally

greater susceptibility of the target organ

higher concentration of active compound

Liver--high blood flow, oxidative reactions

Kidney--high blood flow, concentrates chemicals
Lung--high blood flow, site of exposure
Neurons--oxygen dependent, irreversible damage
Myocardium--oxygen dependent
Bone marrow, intestinal mucosa--rapid divide

Target Sites:
Mechanisms of Action
Adverse effects can occur at the level of the molecule, cell,
organ, or organism
Molecularly, chemical can interact with
Proteins Lipids
DNA
Cellularly, chemical can

interfere with receptor-ligand binding

interfere with membrane function
interfere with cellular energy production
bind to biomolecules
perturb homeostasis (Ca)

Metabolism:
adverse effect depends on the concentration of
active compound at the target site over time
The process by which the administered chemical (parent
compounds) are modified by the organism by enzymatic
reactions.
1o objective--make chemical agents more water soluble and
easier to excrete
decrease lipid solubility
--> decrease
amount at target
increase ionization
--> increase
excretion rate --> decrease toxicity
Bioactivation--Biotransformation can result in the formation of
reactive metabolites

Biotransformation (Metabolism)
Can drastically
effect the rate of
clearance of
compounds
Can occur at any
point during the
compounds
journey from
absorption to
excretion

Compound
Ethanol

Without
Metabolism
4 weeks

With
Metabolism
10mL/hr

Phenobarbital 5 months

8hrs

DDT

Days to weeks

infinity

Biotransformation
Key organs in biotransformation
LIVER (high)
Lung, Kidney, Intestine (medium)
Others (low)

Biotransformation Pathways
* Phase I--make the toxicant more water soluble
* Phase II--Links with a soluble endogenous agent
(conjugation)

Individual Susceptibility
--there can be 10-30 fold difference in
response to a toxicant in a population
Genetics-species, strain variation, interindividual
variations (yet still can extrapolate between
mammals--similar biological mechanisms)

Gender (gasoline nephrotox in male mice only)

Age--young (old too)
underdeveloped excretory mechanisms
underdeveloped biotransformation enzymes
underdeveloped blood-brain barrier

Individual Susceptibility
Age--old
changes in excretion and metabolism rates, body
fat

Nutritional status
Health conditions
Previous or Concurrent Exposures
additive
synergistic

--antagonistic

Excretion:
Toxicants are eliminated from the body
by several routes
Urinary excretion
water soluble products are filtered out of the blood by the
kidney and excreted into the urine

Exhalation
Volatile compounds are exhaled by breathing

Biliary Excretion via Fecal Excretion

Compounds can be extracted by the liver and excreted into
the bile. The bile drains into the small intestine and is
eliminated in the feces.

Milk

Sweat

Saliva