TOXICOLOGY

Setyawati S Karyono
Laboratorium Farmakologi
Fakultas Kedokteran Universitas Brawijaya
 TOXICOLOGY
 Multi-disciplinary application of Scientific
knowledge to the study of toxins and their effects
on people, animals, wildlife and the environment.
 The study of the detection, occurrence, properties,
effects, and regulation of toxic substances
(toxicant)
 any agent capable of producing a deleterious
response in a biological system (a toxic
substance)
the inherent toxicity of a substance, based on
appropriate animal models or information from
human studies
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)
TOXICOLOGY SPECIALIZE

 Toxicology is the most diversified of all

Scientific disciplines, so Toxicologists usually
specialize in some aspect of toxicology e.g
 Medical Toxicology/ Clinical Toxicology: diagnosis
and treatment of human diseaeses caused by
poisons
 Veterinary Toxicology: diagnosis and treatment of
diseases of domesticated and wildlife caused by
poisons
 Food toxicology is the study of the nature,
properties, effects and detection of toxic
substances in food and their disease
manifestation in humans
 Forensic Toxicology: deals with the legal and
medical aspects of poisons in people and
animals;
 Environmental Toxicology- deals with effects of
pollutants on the environment and wildlife
 TOXICOLOGICAL TERMS

 Toxin /Toxicant : A substance that has been

shown to present some significant degree of
possible risk when consumed in sufficient
quantity by humans or animals
 Natural/Inherent :Toxicants occur in Foods as
a result of biosynthetic origin
 Safety: is the absence of evidence of toxicity

 Safety is relative and there is no absolute

safety
 Toxicity is ability to cause harm/adverse effect
 A toxin/poison is difficult to define as many
substances present in food would have
adverse/toxic effects if taken in large/sufficient
dose
 Thus there are toxic and non toxic doses for any
substance
 Frequency-response curve: a plot of the % of
individual with specific response as a function
of dose
DOSE (OF TOXICANT)
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
 Bioconcentration (also called
bioaccumulation or biomagnification)

BCF measures the tendency for concentration for a given substance

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-Response Relationship:
As the dose of a toxicant increases,
so does the response.
4

RESPONSE

0-1 NOEL
2-3 Linear Range 3
4 Maximum Response

0 1 DOSE
DOSE DETERMINES THE BIOLOGICAL RESPONSE
ACUTE TOXICITY

 Acute toxicity: toxic response ,often immediate,

induced by single exposure.
 The acute toxicity of a substance is defined by
its LD50 / lethal dose that will kill 50% of a
group of exposed animals
 LD50 is also used to LD50 (/Kg substance
determine the level body
of toxicity weight
200mg Caffeine

100ng Botulinum
toxin
40g Sodium
chloride
TOXICITY NOMENCLATURE
LD50
Extremely toxic < 1 mg/kg

Highly toxic 1 - 50 mg/kg

Moderately toxic 51 - 500 mg/kg

Slightly toxic 501 - 5000 mg/kg

Practically non-toxic > 5000 mg/kg

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
THERAPEUTIC AND TOXIC EFFECTS
ARE DOSE-RELATED: PHENOBARBITAL

Sleep Death

ED50 LD50

Dose of Phenobarbital
THERAPEUTIC INDEX (TI)

TI (preclinical) LD50 letal dose

ED50

clinical TI TD50 toxic dose

ED50
LD50 COMPARISON

Chemical LD50 (mg/kg)

Ethyl Alcohol 10,000
Sodium Chloride 4,000
Ferrous Sulfate 1,500
Morphine Sulfate 900
Strychnine Sulfate 150
Nicotine 1
Black Widow 0.55
Curare 0.50
Rattle Snake 0.24
Dioxin (TCDD) 0.001
Botulinum toxin 0.0001
CHRONIC TOXICITY

 Chronic toxicity: Toxic effect that requires some

time to develop, e.g cancer.
 Testing for chronic toxicity involve continuous
feeding of the test substance to rodents for 20-
24 months.
 By analogy to LD50, the amount of a
carcinogen required to induce cancer in 50% of
a group of exposed animal is referred to as
TD50
Protocols have been established for safety
evaluation of foods and substances used in
foods :
 Acute toxicity

 Genetic toxicity

 Subchronic

 Chronic

 Teratogenesis
 Traditional high dose feeding of a few
rats/other rodents
 A safety margin of a 100-fold is usually applied
from the NOAEL (no observable adverse effect
level) to determine safe dose for man, i.e the
ADI (acceptable daily intake).
 New approaches to safety evaluation may be
required for Genetically modified foods|(GMOs)
 Comparative toxicology concept
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: (TOXICOKINETICS)
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
PATHWAYS
 Toxicokinetics
Locus of Tissue
action reservoirs
“receptors”
Bound Free Bound Free

Systemic
circulation

Absorption Free substance Excretion

Bound Metabolites

Biotransformation
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.
 The first 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

 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)
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
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 --antagonistic
 synergistic
TOXIC AGENT
ASBESTOS-RELATED HEALTH
CONDITIONS
Cancer Non-Cancerous Lung
 Lung cancer Abnormalities:
 Asbestosis
 Mesothelioma
 Pleural plaques
 Other cancers
 Pleural thickening
 Pleural effusion
(fluid) (Can be an
early indication of
mesothelioma)
EXAMPLES OF ASBESTOS USE
 Shingles
 Floor tiles
 Asbestos cement
 Roofing felts
 Insulation & acoustical
products
 Steam pipes, boilers
 “Popcorn” ceilings
 Patching, joint compounds,
textured paint, asbestos
paper tape
 Brakes, clutch facings
 Plastics, fabrics, paints,
paper
ASBESTOSIS
•Scarring and inflammation of lungs
caused by long-term exposure to
asbestos
•Can be disabling or fatal
•Shortness of breath; Persistent
and productive cough; chest
tightness; can lead to heart
failure
•Onset usually 10-20 years after
initial exposure
•Incidence rate increases and
the disease becomes more
severe with increasing dust level
and duration of exposure.
LUNG CANCER
 Smoking increases risk by 12- 20
times
 Asbestos exposure (non-smoker)
increases risk by 5 times
 Smoking & Asbestos Exposure
together increase the risk 50-90 times
• 5-year survival rate in U.S. only 14%

• Accounts for 50% of all asbestos-

related disease
 Asthma
Reversible obstruction of the upper airways
that causes breathing difficulty

Increasing O3 and Particulate Matter are

associated with
 in asthma symptoms and


medication use
  emergency department visits
  in asthma hospitalizations

 7.7% of adults have asthma ~190,000 adult Oregonians with asthma

7.5% of US children have asthma. If prevalence among Oregon
children is the same, then estimated ~50,000 children in Oregon have
asthma
LEAD Health Effects
• Brain damage and lower
intelligence
• Nervous system damage
• Behavior and learning problems
• Impaired speech and language
• Slowed growth and development
• Kidney and liver damage
• Coma, convulsions, or death
• Detection: Blood Test

About 1 in 22 children in America have

high levels of lead in their blood.
LEAD IN HOMES
 MERCURY
 Brain damage  Lung damage
 Kidney damage  Effects passed
from mother to
 Changes in brain fetus: mental
function, such as retardation,
tremors, changes in blindness,
vision or hearing, seizures, inability
memory problems, to speak
or irritability.  Nausea, vomiting,
 Increased blood diarrhea, skin
pressure or heart rashes
rate  Detection: Urine
Test

 MINAMATA DISEASE
MINAMATA
DISEASE

ADULT MEHG
PESTICIDE EXPOSURE
Health symptoms:
headache, muscle
twitching, tremors, lack of
coordination, abdominal
cramps, diarrhea, nausea,
vomiting, dizziness, chest
tightness, blurred vision,
excess sweating,
weakness.
FAS CHILD
TERATOGENIC EFFECT
FOOD TOXICOLOGY
FOOD TOXICOLOGY

 Food toxicology is the study of the nature,

properties, effects and detection of toxic
substances in food and their disease
manifestation in humans
FOOD POISONING

 Food can have potentially toxic directly

added to it
 Food can be the vehicle for toxic substances

 Food can be the media for the growth of

organisms that produce toxic substances
 The organisms used as food can produce of
accumulate toxic substances
SUBSTANCES ADDED TO FOOD
 Coloring agents
 Antifoaming agents
 Stimulants
 Caffeine
 Sweetening agents
 Various artificial sweeteners
 Texturing agents
 Calcium acetate
 Curing agents
 Calcium chloride
 Drying agents
 Some may contain cobalt
FOOD IS THE VEHICLE

 Atmospheric deposition
 Heavy metals
 POPs or POCs

 Pesticides

 Industrial chemicals

 Motor vehicle-liked substances

 Metals

 PAHs
FOOD IS THE VEHICLE
 Materials deliberately used in food production
 Pesticides
ADIs
Maximum residue levels (MRLs) established by
regulatory agency
 Animal drugs
 Maximum residue levels
 ADIs
FOOD AS THE MEDIA
 Microbial and Fungal toxins or mycotoxins
 Aflatoxins
 Trichothecenes
 T-2 Toxin
 Diacetoxyscirpenol
 Vomatoxin or deoxynivalenol
 Ochratoxins
 Fumonisins

 Zearalenone
 a resorcyclic lactone
FOOD AS THE MEDIA

 Clostridium perfringens
 Meats contaminated with intestinal contents at
slaughter
 Several different toxins

 Bacillus cerus
 Cerulide us heat stable
FOOD AS THE MEDIA
 Staphylococcus aureus
 Common
 Exfoliativetoxins
 Leukociden
 Enterotoxins

 E coli
 O157:H7
 Shiga-like toxin
SUBSTANCES FROM MICROBIAL GROWTH

 Bacterial toxins
 Botulinus toxin from Clostridium botulina
 Can grow in intestinal tract of infants
 Fermentation of protein such as seal or fish eggs in
plastic
 Blocks the release of acetylcholine from
presynaptic nerve fiber
SUBSTANCES MIGRATING INTO FOOD
 Substances from the package
 Substances from the food handling equipment

 Lubricants from food processing equipment

 PRODUCTS FORMED DURING FOOD
PREPERATION
 Nitrosamines, etc
 Burned meats
 PAHs
 Heterocyclic amines
 Food Safety Toxicology
• All testing is conducted through oral
exposure in surrogate species to evaluate the
potential toxicity of the new animal drug to
humans following oral exposure in food.

•The objective is to define the concentration of

drug substance that produces no effect in the
toxicological assay of greatest relevance to
oral human exposure.

N(o) O(bserved) E(ffect) L(evel) or NOEL

FOOD SAFETY TOXICOLOGY
The appropriate NOEL is divided by a “safety
factor” to obtain an “acceptable daily intake”
(ADI),
ADI  which is the amount of drug residue per
kilogram body weight per day that can be
consumed daily over the lifetime of a human
without harmful effect.