PHAR3004
Toxicology – General Principles
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Peta-Gaye Thomas-Brown
z Outline
Define toxicology and the main subdivisions
Explain basic concepts and common terminologies in toxicology
Classify common poisonous plants
Outline common poisonous animals
Describe toxicology samples
Describe methods of analysis in toxicology
Describe the main mechanisms of toxicity
Discuss common poison treatment/antidotes
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Toxicology
Toxicology is the science of poisons
Source, Properties, Actions, Adverse effects, Detection
and Treatment of the conditions poisons produce
Poisons – Chemical/physical agent that produces
adverse responses in biological organisms
Including – household, environmental, industrial and
pharmacological substances
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Toxicology
Paracelsus (1493-1541)
“All substances are poisons: There is none which
is not a poison. The right dose differentiates a
poison and a remedy.”
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Subdivisions of Toxicology
Clinical Toxicology
Study of toxic effects of various drugs in the body, and the treatment and
prevention of drug toxicity in the population
Forensic Toxicology
Study of medical evidence of poisoning, and tries to establish the extent to
which poisons were involved in human deaths
Environmental Toxicology
Study of the effects of pollutants on organisms, populations, ecosystems, and
the biosphere
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Subdivisions of Toxicology
Occupational Toxicology
Experimental Toxicology
And others e.g. Biochemical, Toxinology
z Toxicology Terminologies
Toxic
A chemical/ physical agent have the characteristic of producing an
undesirable/ adverse health effect
Toxicity
Ability of a toxic agent to cause adverse effects in living organisms
Hazard
Likelihood of injury based on accessibility, packaging or formulation
Risk
The probability that an event will occur based on patient vulnerability
z Toxicology Terminologies
Toxicants
Toxic substances from chemicals
Toxins
Poisonous substances produced within living cells or organisms
(Biological compounds) e.g Venom, ricin, botulism
Carcinogen
Ability to induce cancer or increase its incidence and can affect any
cells or tissues e.g. benzene, vinyl chloride, benzo(a)pyrene
z Toxicology Terminologies
Mutagen
Ability to induce hereditary genetic defects or increase their incidence
and effect on DNA e.g. radiation, nitrosoamines
Teratogens
Ability to induce non-hereditary congenital malformations (birth defects)
or increase their incidence and effect on the growing fetus e.g rubella,
thalidomide, PCBs, dioxins
Toxicology
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Terminologies
Endocrine/ hormonal disruptor
Mimic hormone e.g. PBDE, BPA
Block hormone activities e.g.
Tamoxifen
Directly stimulate or inhibit the
endocrine system
Toxicology
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Terminologies
Endocrine/ hormonal
disruptor
e.g. Tamoxifen (Breast
Cancer treatment)
Toxicology – Source of Poisons
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Chemicals – commonest source e.g. drugs,
corrosives
Plants – hashish, cocaine, poison ivy
Animals – least common but most serious source
e.g. venoms from scorpion, spiders, snakes, wasps
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Toxic Effects - Statistics
1. Approx. 2.1 million reported cases of human toxic exposures per year
Route: Most poisons enter the body orally – 76.2%
Where: 92% of the exposures occurred at home; 14% occurred in
a health care facility; 2% occurred at work
Who: 52.7% occurred in children > 6 years; 40% of the cases
involved children > 3 years
2. 85.9% of poison exposure were unintentional
Suicide intent was present in 7.5% of the cases
Therapeutic errors comprised 7% of exposures
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Toxic Effects - Statistics
3. Fatalities:
59% of fatalities occur in the 20-49 year age group
2.2% of fatalities occurred in children > 6 years
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Most Common Sources of Poisons
Adult Exposures Children Exposures
Analgesics - 13.3% Cosmetics and personal care
products - 13.3%
Sedatives/hypnotics/anti-
psychotics - 9.8% Cleaning substances - 10.5%
Cleaning substances - 9.5% Analgesics - 7.2%
Antidepressants - 8% Foreign bodies - 6.8%
Bites/envenomations (snakes, Plants - 6.6%
scorpions, etc) - 7.9%
Topicals - 6.3%
Alcohols - 5.4%
Cough and cold preparations - 5.3%
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Most Common Sources of Poisons
Fatalities
Analgesics – acetaminophen, Amphetamines and cocaine – 61% of
aspirin/ other salicylates = 72% street drug deaths
84% of the fatalities were Ecstasy - involved in 23 fatalities (17-
intentional 24 age group)
Antidepressants – TCA’s = 69% Heroin - 29% of deaths
Sedatives/hypnotics/anti- Carbon monoxide
psychotics – benzodiazepines
Alcohols
= 36%
Cardiovascular drugs
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Toxic Effects
Local (non-specific) – at the site of exposure
Poison start a harmful effect upon contact with the biological
system
It does not require specific sites or receptors to elicit its effects
E.g. swelling or pain at site
Systemic – affecting the entire body
Poisons affect a system/organ far from the portal of entry
E.g. pulmonary, cardiac or CNS toxicity
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Toxic Effects
Stupefying
e.g. Dathura, Cannabis, Chloral hydrate
Abortifacient
e.g. Oleanders, Croton, Calotropis, Aconite, Ergot, Lead, Arsenic,
Mercury, Potassium permanganate
Cattle poisons
e.g. Oleanders, Calotropis, Organophosphates, Arsenic, Strychnine,
Abrus precatorius (John crow beads)
Arrow poisons
e.g Abrus, Croton, Aconite, Strychnine, Curare (poison darts), Snake
venom
z Common Poisonous Plants
Oleander – Cardiac Dumb Cane – Calcium
glycosides oxalate crystals
Ackee – Hypoglycin
John Crow / Jumbie Beads –
Castor Bean – Ricin
Abrin protein
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Common Poisonous Plants
Capsicum annuum – pepper plant
Spathiphyllum spp. – peace lily
Euphorbia spp. – poinsettia
Phytolacca sp. – poke weed, ink berry
z Classification of Poisonous Plants
Group 1 – Serious poisoning
1. Contain systematically active
poisons
e.g Belladonna (Atropa
belladonna); Deadly
nightshade (Solanum spp)
2. Contain e.g. Atropine
Anticholinergic effects – dry
mouth, blurred vision, dilated
pupils, palpitations,
tachycardia, urinary retention
z Classification of Poisonous Plants
Group 2a Group 2b
1. Contain insoluble calcium oxalate 1. Contain soluble oxalate salts
crystals e.g. Dumb Cane (sodium or potassium)
Burning pain Acute hypocalcemia →
cardiac arrest
Local mucous membrane
Renal damage
swelling + irritation
Other organ damage
secondary to precipitation of
calcium oxalate crystals
Systemic toxicity due to large
amounts ingested
Gastroenteritis
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Classification of Poisonous Plants
Group 3
Mild to moderate GIT irritation
- oral ingestion
Dermatitis – skin (various
toxins)
e.g. Beech (Fagus sylvatica);
Pokeweed (unripe berries)
(Phytolacca americana)
z Poisonous Animals
Venomous snake
(fangs)
Wasp (stinger)
Tetradotoxin from Puffer fish
z Toxicology Samples
Samples – Tissues and bodily fluids obtained at autopsy
1. Blood: Best from femoral and iliac veins
Jugular veins – may be contaminated by reflux from
upper thorax
General body cavity – highly contaminated by intestinal
contents
Heart or great vessels in chest – contaminated by
postmortem diffusion of drugs/alcohol from stomach or
from aspirated vomit
z Toxicology Samples
2. Urine – 20-30 mL in sterile containers without preservatives
3. Faeces – used in heavy metals poisoning such as arsenic, lead,
mercury
4. Gastric material – vomit or stomach contents
5. Organs – most common organ collected is the liver
Bile – helpful for morphine & chlorpromazine
Lungs – in cases of solvents
6. Hair & Nails – Heavy metals poisoning & prolonged use of
opiates
z Methods of Analysis
Qualitative Methods:
1. Colour tests – rapid, easily performed but non-
specific screening test
Ferric chloride test for salicylates – Pink-Purple
Zwikker test for barbiturates – Purple
Formaldehyde-sulphuric acid test for benzodiazepines –
Orange
Mandalin test for opioids - Brown
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Methods of Analysis
Qualitative Methods:
2. Chemical tests e.g. Reinsch test – an initial indicator for the
presence of heavy metals in the biological sample
Antimony
Arsenic
Bismuth
Selenium
Thallium
Mercury
z Methods
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Analysis
Reinsch Test –
Spherical
Globules of
Mercury
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Analysis
Reinsch Test –
Octa and Tetra
Hydral Crystals of
Arsenic
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Analysis
Reinsch Test –
Amorphous
Crystals of
Antimony
z Methods of Analysis
Quantitative Methods:
1. Chromatography – a mobile phase is run across a stationary
phase
Thin Layer Chromatography (TLC): Mobile phase – a mixture of
organic solvents such as chloroform or methanol; Stationary phase –
silica gel spread over a class plate
Gas Chromatography-Mass Spectrometry (GC-MS): Mobile phase –
a carrier inert gas such as helium or nitrogen; Stationary phase – a liquid
High Performance Liquid Chromatography (HPLC): Mobile phase –
liquid solvent; Stationary phase – column packed with solid particles
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Thin Layer Chromatography
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zThin
z Layer
Chromatography
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Thin Layer
Chromatography
z Thin Layer Chromatography
1. Samples to be analyzed are spotted near the bottom of the plate and allowed to dry
2. Plate is placed upright in a chamber with the bottom (spotted with sample) placed in
contact with the mobile phase. Mobile phase will move up stationary phase by capillary
action
3. Solvent will move over sample and separate different components of the sample based
on the affinity of the component for the mobile or stationary phase
4. When solvent front (leading edge) reaches the top, the plate is removed and location of
the sample components visualized (using fluorescent dyes and UV light)
5. Result can be quantified by using the Retention Factor (Rf) = distant moved by
sample distant moved by solvent
6. Sample component can be identified by comparing the calculated Rf with Rf of known
substances using a Rf table, given according to mobile and stationary phases used.
z Gas Chromatography – Mass
Spectrometry
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Gas Chromatography
There are 2 types of columns used in GC:
Packed Column – the stationary
phase liquid is coated onto particles
packed into a stainless steel or glass
column
Capillary Column - the stationary
phase liquid is coated onto the walls
of the column itself, which is narrow
and made of glass
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Gas Chromatography
1. Samples are injected into a heated port, where
they are vaporized and carried into the column
with the carrier gas
2. A detector then produces a signal as the sample
components exit the column. The detector is
usually hooked up to a recorder that produces a
gas chromatogram (plot of electronic signal vs
time, which shows a series of peaks that
corresponds to the components in the sample)
3. The retention time (Rt) – time taken for the sample
components to pass through a column, can be
compared to standards to identify the substance The area under each peak is proportional
to the concentration of that substance
Mass Spectrometry
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1. As the sample components exit the GC
column, they are routed into a vacuum
chamber in the mass spectrometer, where
they are hit with a beam of electrons
2. The beam knocks electrons off the sample
component, creating positive electrons and
breaking them into fragments
3. Fragments then pass through an
electromagnetic field, which separates them
by their mass/charge ratio
4. The resulting spectrum plotting the abundance
and mass/charge ration of each fragment is
specific for a given substance
5. This allows definitive identification of the
sample component
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High Performance Liquid
Chromatography
High Performance Liquid
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Chromatography
1. As the mobile phase is pumped through
the column (under pressure), the sample is
injected
2. The sample separates into its components
in the column
3. A detector then identifies the components
as they exit the column, based on their
retention time (Rt)
4. Results are compared to standards
z Methods of Analysis
Quantitative Methods:
2. Immunoassays
Enzyme-multiplied immunoassay technique (EMIT) - used to
detect certain drugs in urine
Fluorescence Polarized immunoassay (FPIA)
Radioimmunoassay (RIA)