Acute Toxicity

AMMAR SARWAR (PHARM-D)

DEFINITION OF TOXICOLOGY
Toxicology can be defined as that branch of science that deals with poisons. In practice, however, many complications exist beyond these simple definitions, both in bringing more precise meaning to what constitutes a poison and to the measurement of toxic effects.

DEFINITION OF TOXICOLOGY

CONTI

Broader definitions of toxicology, such as the study of the detection, occurrence, properties, effects, and regulation of toxic substances, although more descriptive, do not resolve the difficulties. Toxicity is rather a cascade of events starting with exposure, proceeding through distribution and metabolism, and ending with interaction with cellular macromolecules (usually DNA or protein) and the expression of a toxic end point.

TYPES OF TOXICITY
Acute Toxicity: Occuring within 24 hours. Sub-Acute Toxicity: Occuring within 72 hours. Chronic Toxicity: After Years. Sub Chronic: Toxicity: Within 1-3 Months.

TYPES OF TOXICITY
Acute Toxicity: Occuring within 24 hours. Sub-Acute Toxicity: Occuring within 72 hours. Chronic Toxicity: After Years. Sub Chronic: Toxicity: Within 1-3 Months.

CHRONIC TOXICITY

Definition: Toxicity elicited by prolonged exposure and results in delayed effects

1- EXAMPLE OF ACUTE TOXICITY

Mortality occurring within two days of a single dose of a chemical would be a prime example of acute toxicity.

2- EXAMPLE OF CHRONIC TOXICITY

Reduced litter size following continuous (i.e., daily) dosing of the parental organisms would be indicative of chronic toxicity.

(Litter: the offspring at one birth of a multiparous animal <a litter of

puppies>)

ASPECTS OF ACUTE TOXICITY

1- Acute Effect 2- Acute Exposure

ASPECTS OF CHRONIC TOXICITY

1- Chronic Exposure 2- Delayed Effects

3- EXAMPLE

chronic exposure to a persistent, lipophilic chemical may result in sequestration of significant levels of the chemical in adipose tissue of the organism with no resulting overt toxicity. Upon entering the reproductive phase, organisms may mobilize fatty stores, releasing the chemical into the blood stream resulting in overt toxicity including death.

4- EXAMPLES

Acute exposure during a susceptible window of exposure (i.e., embryo development) may result in reproductive abnormalities and reduced fecundity once the organism has attained reproductive maturity.

ACUTE TOXICITY

Acute Exposure

Acute Effects

Time
(a) Short-term exposure resulting in immediate effects

CHRONIC TOXICITY

Chronic Exposure
Chronic Effect

Time
(b) Continuous exposure resulting in sub-lethal effects

MIXED ACUTE/CHRONIC TOXICITY

Chronic Exposure
Acute Effects

Time
(c) Continuous exposure resulting in acute effects

MIXED ACUTE/CHRONIC TOXICITY

Acute Exposure Chronic Effect

Time
(d) Short-term exposure resulting in later sublethal effects

RELATIONSHIP BETWEEN DOSE AND TOXICITY

As the dose of a chemical is increased, the response also increases..

REGULATORY AGENCIES IN THE USA

1-Food and Drug Administration (FDA) 2-Food, Drug, and Cosmetic Act 3-Labor Department 4-Occupational Safety and Health Act 5-Consumer Product Safety Commission 6-Consumer Product Safety Act 7-Environmental Protection Agency (EPA) 8-Federal Insecticide, Fungicide, and Rodenticide Act 9-Clean Air Act

REGULATORY AGENCIES IN THE USA CONTI

10-Federal Water Pollution Control Act 11-Safe Drinking Water Act 12-Toxic Substances Control Act 13-Resource Conservation and Recovery Act 14-State governments 15-Various state and local laws 16-Enforcement of certain aspects of federal law delegated to states

TOXICITY TESTING
Toxicity

assessment; is the determination

of the potential of any substance to act as a poison, the conditions under which this potential will be realized, and the characterization of its action.

CONTROVERSIES.

Among the many areas of controversy are the use of animals for testing and the welfare of the animals, extrapolation of animal data to humans, extrapolation from high-dose to lowdose effects, and the increasing cost and complexity of testing protocols relative to the benefits expected.

TOXICITY TESTING IN ANIMALS

Traditionally the basis for the determination of toxicity has been administration of the test compound, in vivo, to one or more species of experimental animal, followed by examination for clinical signs of toxicity and/or mortality in acute tests. In addition pathological examination for tissue abnormalities is also performed, especially in tests of longer duration.

EXTRAPOLATION OF RESULTS

The results of these tests are then used by a variety of extrapolation techniques to estimate hazard to humans.

TOXICITY TESTING IN ANIMALS

A.

Acute

1. LD50 and LC50oral, dermal or inhaled 2. Eye irritation 3. Dermal irritation 4. Dermal sensitization
B.

Sub-chronic

1. 30- to 90-day feeding 2. 30- to 90-day dermal or inhalation exposure

TOXICITY TESTING IN ANIMALS CONTI

C.

Chronic/reproduction

1. Chronic feeding (including oncogenicity tests) 2. Teratogenicity 3. Reproduction (multi-generation)

D.

Special tests

1. Neurotoxicity 2. Potentiation 3. Metabolism 4. Pharmacodynamics 5. Behavior

ROUTES OF ADMINISTRATION
The nature and degree of the toxic effect can be affected by the route of administration. This may be related to differences at the portals of entry or to effects on pharmacokinetic processes. In the latter case, one route (e.g., intravenous) may give rise to a concentration high enough to saturate some rate-limiting process, whereas another (e.g., subcutaneous) may distribute the dose over a longer time and avoid such saturation.

VARIATION IN TOXICITY BY ROUTE OF EXPOSURE

Chemical Species/Gender Route LD50 (mg/kg)

N-Methyl-N-(1naphthyl) fluoroacetamidea

Mouse/M

Oral Dermal Subcutaneous

371 402

250
115

Rat\M

Oral
Dermal Subcutaneous

300
78

Chlordane

Rat\M

Oral
Dermal

335
840

ROUTES OF ADMINISTRATION CONTI..

1- Oral:

Oral administration is often referred to as administration per os (PO). Compounds can be administered mixed in the diet, dissolved in drinking water, by gastric gavage, by controlled-release capsules, or by gelatin capsules.

ROUTES OF ADMINISTRATION CONTI..

Dermal:

Dermal administration is required for estimation of toxicity of chemicals that may be absorbed through the skin. Compounds are applied, either directly or in a suitable solvent, to the skin of experimental animals after hair has been removed.

ROUTES OF ADMINISTRATION CONTI..

Inhalation: Inhalation toxicity studies are conducted in inhalation chambers. The complete system contains an apparatus for the generation of aerosol particles, dusts, or gas mixtures of defined composition and particle size and a sampling apparatus for the determination of the actual concentration within the chamber.

Injection: Except for certain pharmaceuticals and drugs of abuse, injection (parenteral administration) does not correspond to any of the expected modes of exposure.

ACUTE TOXICITY TESTS

Acute toxicity test methods measure the adverse effects that occur within a short time after administration of a single dose of a test substance. This testing is performed principally in rodents and is usually done early in the development of a new chemical or product to provide information on its potential toxicity.

ACUTE ORAL TESTING

Acute Oral Testing focuses on determining the dose that kills half of the animals (i.e., the median lethal dose or LD50), the timing of lethality following acute chemical exposure, as well as observing the onset, nature, severity, and reversibility of toxicity.

LD50

LD50: (Lethal Dose 50): Amount of a chemical that is expected to kill 50% of the test population LD50 is the measure generally used when exposure is by swallowing, through skin contact, or by injection. Expressed in milligrams of chemical per kilogram of body weight (mg/Kg) of the test animal.

TEST SYSTEM/ANIMAL SYSTEM

2 species required.

Mice rats sometimes rabbits or dogs

DOSE ADMINISTRATION

At least two routes are selected

Oral Dermal
Inhalation

ACUTE ORAL TOXICITY TESTING

Fixed Dose Procedure Principle: lethal doses are avoided

the dose expected to produce some signs of toxicity without causing severe toxic effects or mortality Sighting study: 1 animal at 5/ 50/ 300/ 2000 (limit test)/ (5000) mg/kg bw for dose selection Main study: 4 additional animals at same dose Further groups of animals may be dosed at higher or lower fixed doses, depending on the presence or absence of signs of toxicity or mortality

ACUTE ORAL TOXICITY TESTING

Acute Toxic Class Method Principle: less animals needed

CONT..

a stepwise procedure with the use of 3 animals of a single sex (females) per step, at 5/ 50/ 300/ 2000 (limit test) . Depending on the mortality and/or the moribund status of the animals, on average 2-4 steps may be necessary to allow judgement on the acute toxicity of the test substance.

ACUTE ORAL TOXICITY TESTING

UP

CONT..

and Down Procedure: The up-and-down procedure employs sequential dosing, using only a single animal at each step, the dosage depending on whether the previously dosed animal lives or dies. The test provides a point estimate of lethality and confidence intervals, and can be used to evaluate lethality up to 5000 mg/kg.

EYE IRRITATION
Draize test: The preferred experimental animal is the albino rabbit. The test consists of placing the material to be tested directly into the conjunctival sac of one eye, with the other eye serving as the control.

ACUTE DERMAL TOXICITY STUDY

ACUTE DERMAL TOXICITY STUDY

Limit test at 2000 mg/kg bw: for minimal 10% of body surface (back) fur is removed - solids: moistened with water/ vehicle - liquids: undiluted - test substance should be held in contact with skin for 24 h with porous gauze (=semi-occlusive) to: ensure exposure/ contact prevent ingestion of test substance

ACUTE INHALATION TOXICITY STUDY

ACUTE INHALATION TOXICITY STUDY

groups of animals are exposed for a defined period to the test substance in graduated concentrations, one concentrationbeing used per group - by using inhalation equipment - by using chambers - exposure between 4 and 24 hours as constant as possible - particle size analysis of aerosol concentrations - The LC50 value is expressed as weight of test substance per standard volume of air (mg/1), or as parts per million (ppm).

PARAMETERS

Mortality Clinical pathology Weight change Signs of toxicity

CLASSIFICATION AND LABELLING

It is classified in to five categories of severity where category 1 requires the least amount of exposure to be lethal and
category 5 requires the most exposure to be lethal.

LC50

LC50: (Lethal Concentration 50)

is the concentration of a chemical that expected to kills 50% of a sample population.

LC50 is generally used when exposure to a chemical is through the animal breathing it in,
Normally expressed as milligrams of substance per liter (mg/L) of air or water (ppm)

RELATIONSHIP BETWEEN LD50/LC50 AND TOXICITY

Higher LD50 or LC50

Higher Toxicity

Lower Toxicity

Lower LD50 or LC50

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YOU!