Principle of toxicology; jwliao

Principle of Toxicology
z Introduction
z Classification of Toxicants
z Characteristics of Exposures
Route and Site Exposure
Duration and frequency of Exposure
z Undesired Effects
z Interaction of Chemicals
z Dose Response
Dose Response Relationship
Comparison of Dose Response

z Variation in Toxic Responses

z Descriptive Animal Toxicity Tests
z Historical Control Incidence in Lab. Animals

Introduction
Toxicology:
Study the adverse effects of toxicants in living organisms, and
examine the toxic effects:
1. cellular
2. biochemical
3. molecular mechanism of action
4. occurrence
5. risk assessment

Areas of Toxicology:
Three main categories:
1. Descriptive toxicology
Direct conduct testing, and provide information for safety
evaluation and regulation
e.g., acute oral toxicity test, LD50 levels
2. Mechanistic toxicology
Identifying and understanding the mechanisms of toxicants exert
toxic effect on living organisms
e.g., organophosphate pesticides inhibit AChE
3. Regulatory toxicology
The responsibility for deciding whether a drug or chemical poses
a low risk to be marked
1) Environmental Protection Agency (EPA)
Chemicals and other toxicants
2) Department of Health (DOH)
Food and Drugs
3) Taiwan Agricultural Chemicals and Toxic Substances
Research Institute (TACTRI )
Pesticides regulation in Taiwan
4) Occupational Safety and health Administration (OSHA)
Safety and health in the workplace

Four specialized areas of toxicology

1. Forensic toxicology
Forensic toxicology is a hybrid of analytic chemistry and fundamental
toxicology
Concerned with the medicolegal aspects of the harmful effects of
chemicals on human and animals
To aid in establishing the cause of death in a postmortem
investigation
2. Clinical toxicology
Clinical toxicology is concerned with disease caused by or uniquely
associated with toxic substances
Physicians
Special training in emergency medicine and poison management
3. Environmental toxicology
Environmental toxicology focuses on the impacts of chemical
pollutants in the environment on biological organisms
the effects on human health
non human organisms, fish, birds and terrestrial animals
4. Ecotoxicology
Ecotoxicology is a specialized area within environmental toxicology
The impacts of toxic substances on population dynamics in an
ecosystem
The transport, fate, and interactions of chemicals in the
environmental and ecotoxicology

Classification of toxic agents

1. Spectrum of toxic dose
z Poison
Any agent capable of producing a deleterious response in a
biological system, seriously injuring function or producing death
z Paracelsus (1493-1541)
What is there that is not poison?
All things are poison and nothing [is] without poison
Solely the dose determines that a thing is not a poison

2. LD50
The dosage of chemical needed to produce death in 50 percent of
treated animals, was not a constant

Table 2-1. Approximate Acute Oral LD50 of Some Representative

Chemical Agents
Agents
LD50 mg/kg body weight
Ethyl alcohol
10,000
Sodium chloride
4,000
Ferrous sulfate
1,500
Morphine sulfate
900
Phenobarbital sodium
150
Picrotoxin
5
Strychnine sulfate
2
Nicotine
1
d-Tubocurarine
0.5
Hemicholinium-3
0.2
Tetrodotoxin
0.1
Dioxin (TCDD)
0.001
Botulinum toxin
0.00001

3. Classification
WHO, USEPA, OECD,COA
COA classification of acute toxicity rank of pesticide in mammals

LD50
I. Extremely II .Highly III. Moderately IV. Slightly
(mg/kg) hazardous hazardous
Hazardous
hazardous
()
()
()
()

(Classification)

Solid
Liquid

<5
<20

5-50
20-200

50-500
200-2000

>500
>2000

Solid
Liquid

<10
<40

10-100
40-400

100-1000
400-4000

>1000
>4000

Solid, liquid

<0.05

0.05-0.5

0.5-2

2-20

(Rat)

(Rat)

(Rat) LC50
mg/L

Classification of relative toxicity of general compounds

Volume Dose

Class

Toxicity

Super toxic

1mg / kg

0.004 teaspoon 0.09 teaspoon

Extremely toxic

5mg / kg

0.04 teaspoon

1 teaspoon

Highly toxic

1-50 mg / kg

0.2 teaspoon

4.5 teaspoon

Moderately toxic

50-500 mg / kg 2 teaspoon

1 cup

Slightly toxic

0.5-5 gm / kg

0.45 cup

2.5 quarts

Practically nontoxic 5-15 gm / kg

1.34 cup

2 gallons

Relatively harmless 15 gm /kg

1.34 cup

2 gallons

Dog ( 20 kg )

Cow (450 kg )

Characteristics of exposure
1. Many chemicals are of relatively low toxicity in the native form, but
when acted on by enzymes and interfere with normal cells in the body
2. Influence toxicity related to the exposure, routes, duration and
frequency

z The relationship between elimination and frequency of exposure

 Route and site of exposure

1) Oral GI tract, major route
2) Inhalation Lungs
3) Topical, percutaneous, or dermal Skin
4) Ocular, conjunctiva, cornea Eyes
5) Other routes IP, SC, IM, ID, IV..

Duration and frequency of exposure

Four categories:
1. Acute:
Acute exposure is defined as exposure to a chemical for less than
24 hours
A single administration, repeated exposure may be within a 24-h
period for some slightly toxic or practically nontoxic chemicals
Half of lethal dose (LD50, mg/kg body weight)
2. Subacute: Repeated exposure for 1 month or less
NOAEL (no observed adverse effect levels, mg/kg/day)
3. Subchronic: Repeated exposure for 1~3 months
NOAEL (no observed adverse effect levels, mg/kg/day)
4. Chronic (long term, carcinogenicity): Repeated exposure for more
than 3 months
NOAEL (no observed adverse effect levels, mg/kg/day)
ADI (acceptable daily intake, mg/kg/day) =
NOAEL/(UF*MF)
RfD (Reference dose, mg/kg/day) = NOAEL/(UF*MF)

Spectrum of undesired effects

1. Allergic reactions:
Chemical allergy is an immunologically mediated adverse reaction to
a chemical resulting from previous sensitization or hypersensitivity:
1) Low doses and repeated exposures of chemicals
2) Chemicals combine with an endogenous protein to elicit and
allergy reaction called a hapten
3) The formation of antibodies at least 1 or 2 weeks required
4) Skin Acquired contact dermatitis (ACD), urticaria and itching
5) Eyes conjunctivitis
6) Asphyxia bronchiolar constriction

2. Idiosyncratic reaction
Chemical idiosyncrasy refers to a genetically determined abnormal
reactivity to a chemical
1) Individuals, extreme sensitive to low doses or extreme insensitive
to high doses
2) Succinylcholine
Form pseudocholinesterase, less break down succinylcholine

3. Immediate vs. Delayed toxicity

z Ops inhibited AChE
Respiratory failure after intoxication
z Triothocresylphosphate (TOCP) poison:
Delayed neurotoxicity is not observed until at least several days
after exposure
4. Reversible vs. irreversible toxic effects
Degeneration, regeneration or necrosis, apoptosis
5. Local vs. systemic toxicity
Target organs: hepatoxicity
CNS depression; multiple organic toxicities

Interaction of chemicals
1. Additive effect
The combine effects of two chemicals is equal to the sum of the
effects of each agent given alone
1) Example: 2 + 3 = 5
2) Organophosphous pesticides Cholinesterase inhibition

2. Synergistic effect
The combine effects of two chemicals are greater to the sum of the
effects of each agent given alone
1) Example: 2 + 2 = 20
2) Carbon tetrachloride and ethanol induced hepatoxicity

3. Potentiation effect
One chemical dose not has a toxic effect on a certain organ but when
added another chemical
1) Example: 0 + 2 = 10
2) Isopropanol is not hepatoxic, but enhance carbon tetrachloride
induced hepatoxicity

4. Antagonistic effect
The combine effects of two chemicals interfere with each others
action
1) Example: 4 + 0 = 1
2) Dimercaprol (BAL) chelates with metal ions, As, Pb.
3) Atropine and PAM are OP antidotes
4) Metal-binding protein, Metallothionein binding with Cd

Dose response
Definition:
The characteristics of exposure and the spectrum of effects come
together in a correlative relationship customarily referred to as
the dose-response relationship
Two types:
1. Individual to varying doses of a chemical (Fig. 2-3)
2. Population of individuals (Fig. 2-4)

Evaluating the dose-response relationship

LD50:
The LD50 is the statistically derived single dose of a chemical that
can be expected to cause death in 50% of the animals test
LD50 is statistically calculated by the probit analysis
A sigmoid dose-response curve (Fig. 2-4)
A stated all-or none response is called threshold(Fig. 2-6)

Quantal all-or-none response:

1. Dose-effect curve can be constructed for lethality, cancer, liver
injury, and other toxic responses (Fig. 2-7; 2-8)
2. Calculation on the basis of body weight and body surface (Table
2-2)

Comparison of dose response:

1. Therapeutic index (TI): (Fig. 2-9)
1) For drugs use in risk assessment
2) TI is defined as the ratio of the dose required to produce a toxic
effect and the dose needed to elicit their desired therapeutic
response
3) TI = LD50/ED50
4) The higher TI is relative safety
2. Marginal of safety (MOS):
1) For nondrug chemicals use in risk assessment
2) Estimated exposed dose to a human population and the highest
nontoxic dose determined in experimental animals
3) MOS = LD1/ED99

3. Potency vs. Efficacy: (Fig. 2-10)

1) Potency is referred to the range of doses over which a chemical
produces increasing adverse responses
2) Efficacy reflects the limit of the dose-response relationship on the
response to a certain chemical or function

Variation in toxic responses

Selectively toxicity:
A chemical produces injury to one kind of living without harming
another of life
Pesticides for agriculture:
Less toxic to the plant but injure to fungi, insects
Antibiotics
Penicillin, Kanamycin, Cephalocin.
Species differences toxicity:
Aflatoxin induced liver tumor
15 ppb in rats, but 10,000 ppb in mice
Rats expressed with glutathione S-transferase (mYc) and has
highly catalytic to epoxide of aflatoxin
Mice less detoxified to aflatoxin

Individual difference in response:

Hereditary differences
50% of Caucasian population has a gene deletion for enzyme GST
M1
Genetic polymorphism
Smokers lack null allele, increased risk approximately 3 folds

Descriptive animal toxicity tests

Key points of carcinogenicity test

1. Tumors, both benign and malignant, are not uncommon events in
animals even in the absence of exposure to any carcinogen
2. There are numerous different tumor types that develop spontaneously
in both sexes of both rats and mice, but at different rats
3. Background tumors that are common in one species may be
uncommon in another
4. Even within the same species and strain, large gender differences in
background tumor incidence are sometimes observed
5. Even when the general protocols, diets, environment, strain and source
of animals, and other variables are relatively constant background
tumor incidence can vary widely

Historical control of tumor incidence

(Fig. 2-10; 2-11) Rat spontaneous tumor ref