PRINCIPLES OF TOXICOLOGY

Understand these important principles

Dose...
•
• Dose-response relationships...
• Chemical interactions...
• Mechanisms of Toxicity...
• Routes of exposure...
• Exposure/Effect characteristics...
• Hazard
• Potency...
• Absorption...
• Distribution...
• Elimination...
• Accumulation...
• Detoxification

Principles of Toxicology....

1. Dose: is the most important determinant of toxicity....

• frequency: single (acute), several (sub chronic) or long-term (chronic)....

• dose may be administered a variety of ways:

oral (p.o.), intravenous (i.v.), intramuscular (i.m.), subcutaneous (s.c.)

2. Effect (response):

primary assumptions...

• there is a molecular site (or receptor) with which the chemical interacts to
produce a response...
 • production of response is related to the concentration of the compound
at the active site...

• the concentration of the compound at the active site is related to the dose
administered....

3. Relationship between Dose and Effect: response is graphically

illustrated by a dose-response curve, with dose (independent variable) on x-
axis, and the effect (dependent variable) on y-axis...

effect may be quantal

(all-or-none), such as
death....

or continuous (graded),
such as a change in
blood pressure,
respiration rate, hormone
level, change in
biochemical
concentration....

Evaluating Dose-response:

• For purposes of accurate quantitation, typical dose-response curve is

plotted as log dose vs. percent response....
Both are graphs from the same set of experimental data. The log-dose scale
results in a more linear representation of the data, and is more desirable since
we will use the linear, portion of the curve (from approximately 16 to 84%) to
calculate toxic potency.

For acute (short-term) effects, one common method to calculate toxic potency is
by the LD50 which is the statistically-derived single dose of a substance
that can be expected to cause mortality in 50% of the test animals...

From the graph on the next page, in which log dose is plotted against percent
mortality, we are able to determine, by inspection, that the LD50 is
approximately 100 mg/kg...

This sigmoid curve has a

linear portion between
approximately 16 and
84%. These represent
limits of ± 1 standard
deviation (SD) of the
mean of a normally-
distributed population.
Because these data are
from a normally-distributed
population, the curve can
be transformed to more
accurately describe the
data....
 Sigmoidal curve identical
to the above....

The middle panel is a

histogram in which each
bar represents the % of
animals that died at each
dose minus the % that
died at the immediately
lower dose. By definition,
in the normal, gaussian
distribution, the mean ± 1
SD represents 69.3% of
the population, ± 2 SD is
95.5% and ± 3 SD is
99.7% of the population.
Notice that most of the
animals respond to the
LD50. Those that die from
the smaller doses are
termed sensitive (or
hypersusceptible) while
those that require larger
doses are resistant.

The percent units can be

converted to units of
deviation from the mean
of the normal distribution.
These units, called
normal equivalent
deviations (NEDs)
simplify response
calculations. The NED for
50% response is 0; -1 for
 15.9%, +1 for 84.1% and
so on. To avoid the use of
negative numbers, the
system of probability
units (or PROBITS) was
developed. Probits = NED
+ 5. The data on the
bottom panel are plotted in
probits. When the
population is assumed to
be normal, the probit
transformation can be
applied. The curve is
much straighter, making
an accurate LD50
calculation possible.

% response NED PROBIT

0.1 -3 2
2.3 -2 3
15.9 -1 4
50.0 0 5
84.1 +1 6
97.7 +2 7
99.9 +3 8

Characteristics of Exposure and Effect....

Exposure:

1. Acute: exposure for a duration of less than 24 hr; often a single exposure...

2. Subacute: generally refers to repeated exposure for a month or less.

3. Subchronic: exposure duration from between 1-3 months.

4. Chronic: exposure often greater than 3 months. Usually continual daily

dietary exposure. For animal studies, often for a lifetime of the animal.

Effect: the toxic effects observed following single or repeated exposure to a

chemical are often quite different...

1. Acute Effects: rapidly developing, reaching a maximum with severe

symptoms.
(exposure to high doses of CN- will kill within a few minutes..... )

2. Subacute Effects: symptoms generally not as severe, but toxic effects often
same as acute....

3. Chronic Effects: progresses at a slow and varying rate; may be mistaken

for other diseases. Often difficult to determine cause-and-effect unless in
laboratory....

• asbestos-caused cancer may be delayed 20-30 years....

• acute exposure may result in either acute or chronic effects....

(acute exposure to asbestos may lead to cancer....)

• chronic exposure may result in either chronic or acute effects...

(chronic lead exposure may lead to subacute or acute symptoms....)

4. Accumulative Effects: occurs two ways...

• accumulation of toxin: exposure to heavy metals (lead, mercury) that have

long half-lives result in disease due to metal accumulation....

• accumulation of effect: low level exposure to organophosphate pesticides

depresses acetylcholine esterases to a point where symptoms occur....

5. Delayed Effects: effect may occur only after long exposure; agent cannot be
found in blood or tissues. Damage to system already done....

• radiation sickness

Variations in dose response curves...

1. Predictability: slope of curve...

The slope of the dose-

response curve gives a
clue to the variability of
the data. Compounds
 "A" and "B" (right) have
the same LD50 value,
although the shape of
their curves are different.
The slope for "A" is
relatively "flat,” while that
for "B" is steeper. For
"A," a larger change in
dose is required for a
significant change in
response, while a small
change in dose will
cause an effect with "B."
Thus, there is less
inherent variability with a
compound showing a
steeper curve. All things
being equal, the effects
of a compound with
steeper dose-response
curve are more
predictable.

• comparisons of two compounds by their dose-response curves are

meaningful only when their curves are parallel......

2. Potency: position along the x-axis.....

Position along the x-

axis is indicative of
potency. The more
potent compound is on
the left (squares)
because less compound
is needed to produce an
equivalent response
compared to the
compound depicted on
the right (triangles)....
Compounds vary widely in their potency:

LD50 (mg/kg;
Chemical Agent
rat, oral)
ethyl alcohol 10,000
table salt 4000
sodium pentobarbital 150
strychnine 2
tetrodotoxin 0.10
tetrachlordibenzodioxin (TCDD;
0.001
"dioxin")
botulinum toxin 0.00001

3. Margin of safety....

These three curves

demonstrate how desirable
and undesirable effects of
drugs are illustrated. ED is
effective dose (such as
analgesia), TD is toxic dose,
resulting in some sort of
adverse or "side" effect(s),
while LD is lethal dose. The
therapeutic index of this
drug is the ratio of the TD50
(or LD50) and the ED50; the
margin of safety (a more
conservative estimate) is the
ratio of the TD1 and the ED99.

4. Potency vs. Efficacy.....

 From their relative positions
along the x-axis, compound
"A" is more potent than
compound "B." Both "A" and
"B" also reach maximum
efficacy since their effects
both reach the limit of
response (top).

The difference between

potency and efficacy
(bottom). At low doses, "C" is
more potent than
"D." However, "D" has a
greater maximal efficacy than
"C." Higher doses of "C"
cause no increase in effect.

5. Interactions of chemicals:

a. Additive effects....

The effects of two chemicals,

when given together are often
additive, that is, the combined
effect of two chemicals is
predictable based on the known
toxicity of both compounds. No
specific interactions occur...
1+1=2

b. Synergistic effects....
 The combined effect of the
administration of two
compounds may be greater
than the sum of the two effects;
this is called synergism. The
synergist piperonyl butoxide is
added to some insecticides to
greatly increase their toxicity to
insects.
1+ 1 = 10

c. Antagonism....

The toxic effect of a chemical,

A, agonist, can be reduced
when given with another
chemical, B, the antagonist.
Antagonists, are often used
as antidotes. There are
several mechanisms of
antagonism: 1) functional
antagonism: simple
counterbalancing of the toxic
effect (caffeine and
phenobarbital); 2) chemical
antagonism: antagonist
reacts with the toxin to
reduce toxicity (dimercaprol
chelates toxic heavy metals
such as As); 3) receptor
antagonism: antagonist
binds to receptor, (atropine
with organophosphate
insecticides); 4)
dispositional antagonism:
 fate of the toxin is altered
(cholestyramine can prevent
absorption of organic
chemicals by binding with
them).

Classification of toxicants....

In toxicology, compounds are classified various ways, by one or more of the

following classes:

Classification by....

Use: pesticides (atrazine), solvents (benzene), food additives (nutrasweet)

Effects: carcinogen (benzo-α-pyrene), mutagen (methylnitrosamine),

hepatotoxicant (CHCl3)....

Physical state: oxidant (ozone), gas (CO2), dust (Fe2O3), liquid (H2O)....

Chemistry: aromatic amine(aniline), halogenated hydrocarbon (methylene

chloride)....

Mechanism: cholinesterase inhibitor (malathion), methemoglobin producer

(nitrite)....

Mechanisms of Toxicity....read Chapter 3

Target Organ Toxicity: many toxins do not produce general effects but are
specific to only a few organs......

• vinyl chloride: liver cancer

• asbestos: mesothelioma

• paraquat: lung toxicity

• cadmium: kidney toxicity

Target organs are often not the site of the highest concentration of a chemical....
 • Lead concentrates in bone, but its effects are mainly seen in soft tissues,
such as liver, kidney and blood cells....

• DDT accumulates in adipose tissue (fat stores), but produces no effects

there...it is primarily a central nervous system toxin....

Target organs most frequently affected by toxicants:

1. Central nervous system

2. Circulatory system (blood, blood-forming system)

3. Visceral organs (liver, kidneys, lung)

4. Muscle and Bone

Possible mechanisms of tissue sensitivity: i.e., why are toxins often selective
to tissues?

• preferential accumulation: toxicant may accumulate in only certain

tissues, and cause toxicity there....

Cd in kidney, paraquat in lung

• selective metabolic activation: enzymes needed to convert a

compound to the active form may be present in highest quantities in a
particular organ…..

CCl4, nitrosamines in liver

• characteristics of tissue repair: some tissues may be protected from

toxicity by actively repairing toxic damage; some tissues may be
susceptible because they lack sufficient repair capabilities....

nitrosamines in liver

• specific receptors and/or functions: toxicant may interact with

receptors in a given tissue....

curare: a receptor-specific neuromuscular blocker used in dart

poisons

• Physiological sensitivity: the nervous system is extremely sensitive to

agents that block utilization of oxygen....
 nitrite: oxidizes hemoglobin (methemoglobinemia)

cyanide: inhibits cytochrome oxidase (cells not able to utilize

oxygen)

barbiturates: interfere with sensors for oxygen and carbon dioxide

content in blood....