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

 z
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
 z
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

 z
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
1. Contain insoluble calcium oxalate
crystals e.g. Dumb Cane
 Burning pain
 Local mucous membrane
swelling + irritation
Group 2b
1. Contain soluble oxalate salts
(sodium or potassium)
 Acute hypocalcemia →
cardiac arrest
 Renal damage
 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
 z z
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
z Methods
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Analysis

Reinsch Test –
Octa and Tetra
Hydral Crystals of
Arsenic
z Methods
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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
 z

zThin
z Layer
Chromatography
 z

z
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
z z
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
 z z
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)