Principle of toxicology; jwliao

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

Variation in Toxic Responses Descriptive Animal Toxicity Tests 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
Poison Any agent capable of producing a deleterious response in a biological system, seriously injuring function or producing death 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
(Classification) (Rat) (Rat) (Rat) LC50 mg/L Solid, liquid <0.05 0.05-0.5 0.5-2 2-20 LD50 I. Extremely II .Highly III. Moderately IV. Slightly (mg/kg) hazardous hazardous Hazardous hazardous () () () () Solid Liquid Solid Liquid <5 <20 <10 <40 5-50 20-200 10-100 40-400 50-500 200-2000 100-1000 400-4000 >500 >2000 >1000 >4000

Classification of relative toxicity of general compounds Class Super toxic Extremely toxic Highly toxic Moderately toxic Slightly toxic Toxicity 1mg / kg 5mg / kg 1-50 mg / kg 0.5-5 gm / kg Volume Dose Dog ( 20 kg ) 0.04 teaspoon 0.2 teaspoon 0.45 cup 1.34 cup 1.34 cup Cow (450 kg ) 1 teaspoon 4.5 teaspoon 1 cup 2.5 quarts 2 gallons 2 gallons 0.004 teaspoon 0.09 teaspoon

50-500 mg / kg 2 teaspoon

Practically nontoxic 5-15 gm / kg Relatively harmless 15 gm /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

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 Ops inhibited AChE Respiratory failure after intoxication 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