Biochemical Toxicology
Introduction to You Know ?
Biochemical Toxicology
Chatchawin PETCHLERT, Ph.D.
Department of Biochemistry
Faculty of Science, Burapha University
Review
Toxicology is the science that studies the harmful effects of
overexposure to drugs, environmental contaminants, and
naturally occurring substances found in food, water, air, and soil.
– Main objectives are to establish safe doses and determine
mechanisms of biologic action of chemical substances.
A career in toxicology involves evaluating the harmful effects and
mechanisms of action of chemicals in people, other animals, and
all other living things in the environment.
– This work may be carried out in government, private
industry and consulting firms, or universities and other
research settings.
Toxicologists routinely use many sophisticated tools to
determine how chemicals are harmful.
(e.g.) computer simulations, computer chips, molecular
biology, cultured cells, and genetically-engineered
laboratory animals .
Chatchawin PETCHLERT, Ph.D.
07/06/53
92% of all poisonings happen at home.
The household products implicated in most
poisonings are: cleaning solutions, fuels,
medicines, and other materials such as
glue and cosmetics.
Certain animals secrete a xenobiotic poison
called venom, usually injected with a bite
or a sting, and others animals harbor
infectious bacteria.
Some household plants are poisonous to
humans and animals.
History
2700 B.C. - Chinese journals: plant and
fish poisons
1900-1200 B.C. - Egyptian documents
that had directions for collection,
preparation, and administration of
more than 800 medicinal and poisonous recipes.
800 B.C. - India - Hindu medicine includes
notes on poisons and antidotes.
50-100 A.D. - Greek physicians classified over
600 plant, animal, and mineral poisons.
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History History
50- 400 A.D. - Romans used poisons for
executions and assassinations. Swiss physician Paracelsus (1493-1541)
credited with being
The philosopher, Socrates, was executed “the father of modern toxicology.”
using hemlock for teaching radical
ideas to youths. “All substances are poisons: there is none
which is not a poison. The right dose
Avicenna (A.D. 980-1036) Islamic authority on differentiates a poison from a remedy.”
poisons and antidotes.

1200 A.D. - Spanish rabbi Maimonides writes

first-aid book for poisonings,
Poisons and Their Antidotes

History History
Italian physician
Ramazzini (1713) published
“De Morbis Artificum”
(Diseases of Workers)

describing "asthma" in bakers, miners, farmers, gilders, tinsmiths,

glass-workers, tanners, millers, grain-sifters, stonecutters,
ragmen, runners, riders, porters, and professors. Ramazzini
outlined health hazards of the dusts, fumes, or gases that such
workers inhaled. The bakers and horse riders described by Spanish physician,
Ramazzini would today probably be diagnosed as suffering from Mattieu Joseph Bonnaventura Orfila (1815)
allergen-induced asthma. The lung diseases suffered by most of established toxicology as a distinct scientific discipline.
the other workers would now be classified as "pneumoconiosis,"
a group of dust-related chronic diseases. “Toxicology is a study of poisons”

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History History
20th Century 20th Century

Paul Ehrlich –developed staining procedures to observe Rachel Carson - alarmed public about
cell and tissues and pioneered the understanding of how dangers of pesticides in the environment.
toxicants influence living organisms.

Toxicology Toxicology
What is toxicology? The study of the effects of poisons. Toxicology is arguably the oldest scientific discipline, as the
Poisonous substances are produced by plants, animals, or earliest humans had to recognize which plants were safe
bacteria. to eat.
Phytotoxins
Zootoxins
Bacteriotoxins

Most exposure of humans to chemicals is via naturally

Toxicant - the specific poisonous chemical. occurring compounds consumed from food plants.
Xenobiotic - man-made substance and/or produced by but not Humans are exposed to chemicals both inadvertently and
normally found in the body. deliberately.

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Toxicology Biochemical and Molecular Toxicology

• Involves all aspects of the adverse effects of chemicals on living
systems
• Examines the cellular, biochemical and molecular mechanisms of
action as well as functional effects such as neurobehavioral and
immunological, and assesses the probability of their occurrence
• Fundamental to this process is characterized as the relation of
exposure (or dose) to the response
• Much of toxicology deal with the exogenous compounds to the
normal metabolism of the organism, with such compounds being
referred to as xenobiotics. However, many endogenous compounds
are toxic when administered in unnaturally high doses. Similarly,
trace elements, which are essential in the diet at low concentrations,
are frequently toxic at higher levels.
• concerned with the definition, at the molecular and cellular levels,
levels of
the cascade of events that is initiated by exposure to a toxicant and
culminates in the expression of a toxic endpoint
• The tools of chemistry, biochemistry and molecular biology are the
primary tools of toxicology and progress in toxicology is closely
linked to the development of new methodology in these sciences.
• Those of chemistry provide analytical methods for toxicants and their
metabolites, particularly for forensic toxicology,
toxicology residue analysis, and
toxicant metabolism; those of biochemistry provide methods for the
investigation of metabolism and modes of toxic actions; and those of
molecular biology provide methods for investigation of the roles of
gene expression in toxicity.

Mechanistic Toxicology What Do Toxicologists Do?

Most toxicologists work to develop a

How do chemicals cause mechanistic understanding of how
chemicals affect living systems:
their toxic effects?
– Develop safer chemical
products
– Develop safer drugs
– Determine risks for chemical
exposures
– Develop treatments for chemical
exposures
– Teach ( e.g. other toxicologists,
graduate students, and youth)

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What Do Toxicologists Do? What Do Toxicologists Do?

Mechanistic toxicologists study how a chemical
causes toxic effects by investigating its absorption,
distribution, and excretion. They often work in
academic settings or private industries and develop
antidotes.
Forensic toxicologists study the
application of toxicology to the law. They
uses chemical analysis to determine the
cause and circumstances of death in a
? postmortem investigation.

Environmental toxicologists study the

Descriptive toxicologists evaluate the toxicity of effects of pollutants on organisms,
drugs, foods, and other products. They often perform populations, ecosystems, and the
experiments in a pharmaceutical or academic setting. biosphere.

Clinical toxicologists usually are physicians or Regulatory toxicologists use scientific

veterinarians interested in the prevention, diagnosis, data to decide how to protect humans and
and treatment of poisoning cases. They have animals from excessive risk.
specialized training in emergency medicine and Government bureaus such as the FDA
poison management. and EPA employ this type of toxicologist.

Regulatory Toxicology What is a Toxin?

Use data from descriptive and mechanistic toxicology to

perform risk assessments.
The term “toxin” refers to toxic
Concerned with meeting requirements of
regulatory agencies.
substances that are produced
naturally.
Industry/government interactions.

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What is a Toxicant? What are Harmful or Adverse Effects?

The term “toxicant” refers to toxic

substances that are produced by or Those effects which are damaging
are a by-product of human-made to either the survival or normal
activities. function of the individual

Tolerance
Modern Toxicology
Tolerance is a state of decreased responsiveness to a toxic
effect of a chemical resulting from prior exposure to that 1961 - Society of Toxicology
chemical or to a structurally related chemical.

1970s - EPA, FDA, and NIOSH

decreased amount of toxicant reduced
reaching the site where the responsiveness of a
toxic effect is produced tissue to the chemical

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Toxicology Terms Toxicology Terms

Toxicity - The adverse effects Exposure – Contact providing

that a chemical may produce. opportunity of obtaining a
poisonous dose.

Dose - The amount of a chemical

that gains access to the body. Hazard – The likelihood that the
toxicity will be expressed.

Exposure Concepts

Different toxic responses may arise from

different:

– Routes of exposure.

– Frequencies of exposure.

– Duration of exposure (acute vs.

chronic).

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Routes of Environmental Exposure Routes of Environmental Exposure

Ingestion (water and food)

Absorption (through skin)
Injection (bite, puncture, or cut)
Inhalation (air)

Some routes of exposure The Dose Makes the Poison

Intravenous An apparently nontoxic chemical

can be toxic at high doses. (Too
much of a good thing can be
bad).

Highly toxic chemicals can be life

saving when given in appropriate
doses. (Poisons are not harmful
at a sufficiently low dose).

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Fundamental Rules of Toxicology Chemicals, Chemicals Everywhere

Exposure must first occur for the chemical to present Everything in the environment is made of chemicals. Both
naturally occurring and synthetic substances are chemical in
a risk.
nature.

The magnitude of risk is proportional to both the People are exposed to chemicals by
potency of the chemical and the extent of eating or swallowing them, breathing
exposure. them, or absorbing them through the
skin or mucosa.

“The dose makes the poison” (amount of chemical at People can protect themselves by
the target site determines toxicity). blocking these routes of exposure.

Duration & Frequency of Exposure Exposure Concepts

Duration and frequency are also important Exposure to chemicals may come from
components of exposure and contribute to
dose. many sources:
– Environmental
Acute exposure - less than 24 hours;
– Occupational
usually entails a single exposure
– Therapeutic
Repeated exposures – Dietary
are classified as: – Accidental
– Subacute - repeated for up to 30 days – Deliberate
– Subchronic - repeated for 30-90 days
– Chronic -repeated for over 90 days

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Children & Poisons Individual Responses Can Be Different

The variety of responses among organisms that get the
same dose of chemical is due to individual
susceptibility.
Dose and individual susceptibility play roles in all
situations involving chemicals, including those making
medicine and caffeine.

What is a dose?
What is dose-
dose-response?
Threshold Effects for Dose

Dose-Response Is there such a thing

Relationships as a ‘safe’ dose??
The dose is the actual amount of a chemical that enters
the body.
Agent A

Response
Dose-response suggests that a dose, or a time of Agent B
exposure (to a chemical, drug, or toxic substance), will
cause an effect (response) on the exposed organism.
“NOAEL”
(No Observable Adverse
Threshold dose suggests that there should be a dose or Effect Level)
exposure level below which harmful or adverse effects are
not seen in a population.
Dose

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Dose--Response Relationship
Dose Lethal Doses
LD50 is the dosage (mg/kg body weight) causing death in 50%
of exposed animals.
Agents LD50 (mg/kg)*
Ethyl alcohol 10,000
Sodium chloride 4,000
Ferrous sulfate 1,500
Morphine sulfate 900
Phenobarbital sodium 150
Therapeutic Index (TI) = TD50/ED50 Picrotoxin 5
Strychnine sulfate 2
Nicotine 1
d-Tubocurarine 0.5
Hemicholinium-3 0.2
Tetrodotoxin 0.10
Dioxin (TCDD) 0.001
Botulinum toxin 0.00001

Introduction to Xenobiotics How Does the Body Prevent the Actions

of Xenobiotics ?
*Recall: Foreign chemicals are
synthesized within the body are
termed xenobiotics (Gr. Xenos 1) Redistribution
meaning “strange”)*
2) Excretion – (primarily water soluble compounds)
- kidney and liver
• Xenobiotics may be naturally 3) Metabolism – the major mechanism for terminating
occurring chemicals produced by xenobiotic activity, and is frequently the single most
plants, microorganisms, or animals important determinant of the duration and intensity of
(including humans). toxic responses to a xenobiotic.
- liver, kidney, lung, GI, and others
• Xenobiotics may also be synthetic chemicals produced by
humans.
Poisons are xenobiotics, but not all xenobiotics are poisonous.
poisonous. Note: 1) and 2) are highly dependent upon 3)

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Xenobiotics at Work General Scheme of Xenobiotic Metabolism

TOXICOKINETICS
Lipophilic Hydrophilic
(parent compound) (metabolite)
1) Decrease biological activity
Metabolism 2) Increase excretability

Xenobiotic

Phase I Phase II Metabolites

Metabolites
(oxidative) (synthetic)
size
Bioactivation polarity ionization
Detoxification
Detoxification functionality water solubility
Excretion Increase excretability

How Xenobiotics Cause Toxicity Types of Toxic Effects

Some xenobiotics cause toxicity by disrupting normal cell functions: Death - arsenic, cyanide

– Bind and damage proteins (structural, enzymes) Organ Damage - ozone, lead

– Bind and damage DNA (mutations)

Mutagenesis - UV light
– Bind and damage lipids
Carcinogenesis - benzene, asbestos
– React in the cell with oxygen to form
“free radicals” which damage lipid, protein, Teratogenesis - thalidomide
and DNA

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Biochemical Toxicology
Target Organ Toxicity
Central Nervous System – lead
Immune System - isocyanates
Liver - ethanol, acetaminophen
Respiratory Tract - tobacco smoke,
asbestos, ozone
Eye - UV light (sunlight)
Kidney - metals
Skin - UV light, gold, nickel
Reproductive System –
dibromochloropropane
Animals in Research
“Virtually every
medical achievement
of the last century has
depended directly or
indirectly on research
in animals.”
U.S. Public Health
Service
Chatchawin PETCHLERT, Ph.D.
07/06/53
What Is the Risk?
People can make some choices about chemical exposure;
however, some exposure is controlled at a level other than
an individual one. Collective groups of people, such as
communities and governments, seek to control chemical
exposure on a community or global level.
Descriptive Animal Toxicity Tests
Two main principles Test Material Identification
1. The effects produced by a
Chemical Characterization
compound in laboratory
animals, when properly
qualified, are applicable to
Literature Review
humans. Structure/Activity Assessment
2. Exposure of experimental
animals to chemicals in Short-term Animal Studies
high doses is a necessary
(Acute/Short-term Repeated Dose)
and valid method of
discovering possible
In vitro Genetic Toxicology Metabolism/Pharmacokinetics
hazards in humans.
Subchronic Toxicity
Reproductive/Teratology Chronic Toxicity Oncogenicity
Typical tiered testing scheme for the toxicological evaluation of new chemicals 52
(From: Wilson et al., 2001; In: Hayes, 2001; Principles and Methods in Toxicology, 4th ed., Fig.19-1, p. 918.)
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International Conference on Harmonization (ICH) Codification of “Safety”

Protocols. Titles and abbreviations adopted November, 2005. Summary
Carcinogenicity studies
S1A Need for carcinogenicity studies of pharmaceuticals Toxicology is a fascinating science that
S1B Testing for carcinogenicity of pharmaceuticals
S1C(R1) Dose selection for carcinogenicity studies of pharmaceuticals & limit dose makes biology and chemistry interesting
Genotoxicity studies
S2A Guidance on specific aspects of regulatory genotoxicity tests for pharmaceuticals
and relevant.
S2B Genotoxicity: A standard battery for genotoxicity testing of pharmaceuticals
Toxicokinetics and pharmacokinetics
S3A Note for guidance on toxicokinetics: The assessment of systemic exposure in Understanding HOW (i.e. mechanism)
S3B toxicity studies
Pharmacokinetics: Guidance for repeated dose tissue distribution studies something produces a toxic effect can lead to new ways of
Toxicity testing preventing or treating chemically-related diseases. Animal use
S4 Single dose toxicity tests
Duration of chronic toxicity testing in animals (rodent and non rodent toxicity
in research is essential for medical progress.
testing)
Reproductive toxicology
S5(R2) Detection of toxicity to reproduction for medicinal products & toxicity to male Many diseases are the result of an interaction between our
fertility
Biotechnological products
genetics (individual variability) and chemicals in our
S6 Preclinical safety evaluation of biotechnology-derived pharmaceuticals environment.
Pharmacology studies
S7A Safety pharmacology studies for human pharmaceuticals
S7B The non-clinical evaluation of the potential for delayed ventricular repolarization
(QT interval prolongation) by human pharmaceuticals
Toxicology provides an interesting and exciting way to apply
Immunotoxicology studies science to important problems of social, environmental, and
S8 Immunotoxicity studies for human pharmaceuticals public health significance.
Joint safety/efficacy (multidisciplinary) topic
M3(R1) Non-clinical safety studies for the conduct of human clinical trials for
pharmaceuticals Data from: http://www.ich.org/cache/compo/276-254-1.html

Toxicological Information Sources Electronic Databases

• Toxicology Data Network (TOXNET)
• CHEMTREC (Chemical Transportation
• The Agency for Toxic Substances and Disease Registry Emergency Center)
(ATSDR)
• MEDTREC (Medical Transportation
• The National Center for Environmental Health (NCEH) Emergency Center)
• The United States Environmental Protection Agency (EPA) • Material Safety Data Sheets (MSDS)
• The National Institute for Safety and Health (NIOSH) • HazDat (Hazardous Substances and Health
• The Centers for Disease Control and Prevention (CDC) Effects Database)
• The Food and Drug Administration (FDA) • Thailand Chemical Safety Website
• The American Conference of Governmental Industrial • Chemtrack
Hygienists (ACGIH)
• The Nuclear Regulatory Commission (NRC)

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Local Toxicological Information Sources Local Toxicological Information Sources

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International Toxicological Conference

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