Environmental issues and hazards 


in the chemical research laboratory

Chemical toxicology

Ulf Ellervik
normal spider web spider treated with
marijuana

OH
H

O
Chemical Toxicology - outline

1. Introduction
2. Toxicological concepts
3. Toxicity
4. Acute toxicity
5. Toxicological testing
6. Uptake and distribution of chemicals
7. Metabolism - Phase I
8. Metabolism - Phase II
9. Excretion
10.Electrophiles
11.Examples (reference only)
3. Why are chemicals dangerous?
Examples:

O O O
Toxic P Carcinogenic S
N MeO OMe
CN
O

Suffocating CO2 N2 Oxidizing H2O2

S
H2SO4
Burning Cl Cl Narcotic
NH2

Cl
O2N NO2 NC
Explosive N N Irritating
Hg(ONC)2
CN
N
NO2

Flammable CS2 Radioactive T2O

O
1. Introduction

• What is a poison?
Alle Ding sind Gift, und nichts ohn Gift; allein die Dosis macht,
da ein Ding kein Gift ist.

All things are poison and nothing is without poison, only the dose
permits something not to be poisonous.
Paracelsus, 1538

more than 1 liter of water each hour gives serious problems....

 1. Introduction
OH Compoun d Lethal dose
Ethanol 700 g
etanol
NaCl Salt 300 g
Acetyl salicylic acid 100 g
DDT 8 g
COOH
O O
Strycnine 150 mg
Nicotine 100 mg
Arsenic (III) oxide 100 mg
CCl3 Tetrodoto x i n 10 mg CH3-Hg-CH3
Dimethyl mercury 0,1 mg
Cl O Cl
Cl Cl 2,3,7,8-tetrachlorodibenzodioxin e 0,1 mg
Botulinum toxin 0,001 mg Cl O Cl

H
H N As2O3
N H
O
N
O
2. Toxicological concepts
a) Acute or chronic effects

Acute effects are observed immediately or

shortly after an exposure at one single occasion
or multiple doses within 24 h.

Subacute effects means exposure during less

than 1 month

Subchronic means exposure over 1-3 months

Chronic effects are the result of a continous

exposure during at least 3 months
Time
2. Toxicological concepts
a) Acute or chronic effects
example:

acute effect: narcotic

chronic effect: leukemia

2. Toxicological concepts
b) Reversible or irreversible effects

Reversible effects disappear if the exposure stops

-example: narcotic effect of solvents

Irreversible effects never disappear

-example: damages to nerves, tumors

2. Toxicological concepts
c) Local or systemic effects
Local effects are observed on the part of the body
that first came in contact with the chemical

example: acid burns

Systemic effects require tha the chemical is

uptaken and distributed to the target organ

Target organs: central nervous system often

circulatory system
the liver
the kidneys
the lungs
the skin

muscles
bones rarely
2. Toxicological concepts
It is difficult to estimate the total
d) Total effect effect of a mixture of chemicals!

Independent effect - two compounds are toxic independent of each other

2+2=2
Additive effect - the total effect is the sum of the the two independent effects
example: toluene + xylene 2+2=4

Synergistic effect - the united effect is stronger than the additive effect
example: ethanol + carbon tetrachloride 2+2=8

Potentiating effect - one of the compounds is not toxic in itself but enhance
the effect of another compound 0+2=4
example: 2-propanol + carbon tetrachloride

Antagonistic effect - one compound opposes the effect of another 2+2=1

-functional: one raise blood pressure and one lowers it
-chemical: two compounds neutralize each other
-dispositional: one compound lowers the uptake of another
-receptoric: the two compounds bind to the same receptor
3. Toxicity
a) Physical state
The purity of a compound is important to evaluate the
toxicity (impurities can have very strong effects)

The physical state of a compound cam influence the

toxicity:
-vapor or aerosols are easily uptakne by the lungs
-some solids are not easily dissolved and thus not
toxic
-finely ground arsenic oxide is much more toxic
compared to granlulates
-solution can be uptaken by the skin

example: mercury
3. Toxicity
b) Exposure
The route of exposure is very important for the toxicity

intravenous (ivn) - injected in a vein

inhalated (ihl)

intraperitoneal (ipr) - injected in the peritoneal cavity

subcutaneous (scu) - injected into the skin

oral (orl) - swallowed

dermal (d) - applied on the skin

3. Toxicity
b) Exposure
3. Toxicity
c) The victim
-Species-related differences

human???
3. Toxicity
c) The victim
-Sex-related differences
there are usually differences between males
and females
 3. Toxicity
c) The victim
-Age-related differences
in general, infants (undeveloped systems)
and old people (poor immune system)
are more sensitive to toxic chemicals
LD (μg/kg)
50
Soman in rats
140

120

100

80

60

40

20

0
0 50 100 150 200 250

Age (days)
3. Toxicity
c) The victim
-Individual-related differences
 4. Acute toxicity
a) Dose-response relationships
Sum of deaths (%) LD50 -the dose that will kill 50% of the
exposed individuals
LD10 -the dose that will kill 10% of the
exposed individuals
LDlow -the lowest dose known to cause
death
LC50 -the concentration (mg/m3) that will kill
50% of the exposed individuals
TDx -the dose that will give a toxic effect in
X% of the exposed individuals

LD50 log Dose ex. LD50 = 25 mg/kg (or, rat)

economical and ethical issues

(more than 100 animals for a normal test)
4. Acute toxicity
b) specific toxicity
4. Acute toxicity
c) non-specific toxicity
-the toxicity is dependent on the lipophilicity

toxicity

logP
4. Acute toxicity
d) selective toxicity
-the toxicity is selective to one species or one organ
example: penicillin, DDT
5. Toxicological testing
5. Toxicological testing
Risk assesment
-Comparison of the effects on different organisms (animals, cells....)
and men

-Dose-response relationships

-Weighing the risks against use, pleasure etc

5. Toxicological testing
Risk assesment
6. Uptake and distribution
gastro- skin lungs intravenous other
uptake intestinal (mucous
tract membranes)

distribution portal vein circulatory system

liver kidney lung glands interstitial liquid

gall bladder bone other

fat tissue

excretion faeces urine exhaled air secretion

6. Uptake and distribution
6.1 By the skin
Uptake of chemicals by the skin depends on:
-the general condition (brusies etc)
-the thickness (arms thin, palms thick)
-the water content of the epidermis (gloves)

Many compounds enter easily by the skin

(dichloromethane, methanol)

DMSO increase the permeability for other compounds

(used in veterinary medicine)

epidermis

dermis

blood vessels
6. Uptake and distribution
6.2 By the gastro-intestinal tract

mechanical enzymatic
degradation degradation bacterial
chemical
degradation degradation
pH = 2
Total surface 300 m2 (a tennis court)
6. Uptake and distribution
6.2 By the gastro-intestinal tract
The absorption takes place in villi

Toxic compounds sometimes fit into transport systems

examples: -thallium and cobalt fit into the iron transporter

-lead fit into the calcium transporter

Some compounds pass through the system

example: -elemental mercury

6. Uptake and distribution
6.3 By the lungs
-100 m2 (a badminton court)
-the exchange takes place in the alveoli where the distance
between the blood and the air is only 1 μm
6. Uptake and distribution
6.3 By the lungs
Aerosols are particles or droplets that are sufficiently
small to hover in the air
Dust (solid particles)
Fume (particles formed by combustion)
Smoke (particles formed by combustion of
organic material)
Fog, mist (liquid droplets)
Smog (particles from car exhausts)
6. Uptake and distribution
6.3 By the lungs
The uptake is dependent on size
-small particles (< 1 μm) goes down in the alveoli and can
cause e.g. silicosis or asbestosis
-bigger particles are stopped in the mucus membranes
and transported by the mucociliary escalator:
6. Uptake and distribution
6.3 By the lungs
-Hydrophilic gases are usually absorbed by the mucus
membranes

-Very reactive gases (HCl, NH3, SO2) are irritating and

blocks respiration and can therefore not be inhaled in
larger quantities

-Chemicals with intermediate reactivity and lipophilicity

(phosgene, ozone, isocyanates) can be inhaled and give
injuries at all levels in the lungs
They harm the epithelial cells (by lipid peroxidation) so that
these are leaking fluid - pulmonary oedema (internal
drowning) which takes up to 48 h to develop
6. Uptake and distribution
6.4 Bioaccumulation
Chemicals can be concentrated
in organisms (bioconcentration)
for example in fat
Chemicals that are relatively
stable and not rapidly
converted may be absorbed
by another organism that
feeds on the first. (biomagnification)

example: polyhalogenated
aromatic hydrocarbons (PCB)
6. Uptake and distribution
gastro- skin lungs intravenous other
uptake intestinal (mucous
tract membranes)

distribution portal vein circulatory system

liver kidney lung glands interstitial liquid

gall bladder bone other

fat tissue

excretion faeces urine exhaled air secretion

6. Uptake and distribution
6.5 Accumulation
Absorbed compounds are distributed by the blood but are
sometimes accumulated in target organs.

Brain, nerves, fatty tissue: lipophilic compounds

Bones: inorganic lead, barium, strontium, fluorine, tin

Liver: mercury vapour, manganese, organic lead

Kidneys: cadmium, mercury

Thyroid gland: iodine

Hair: Arsenic
6. Uptake and distribution
6.6 Biological barriers
The endothelial cells of the blood capillaries in the brain
are held together and are surrounded by glial cells. This
makes the passage of compounds difficult and constitute
the blood-brain barrier.

Problem: elemental mercury is lipophilic enough to pass

the blood-brain barrier. If it is oxidized to Hg2+ it will not
get out again...

The placenta regulates the flow of chemicals to the fetus

7. Metabolism
Biotransformation is not equal to detoxification!

The metabolism will effect the biological activity of a

compound
7. Metabolism - Phase I
Summary of important Phase I and Phase II reactions
8. Metabolism - Phase II
8.1 Conjugation with sulfate
8. Metabolism - Phase II
8.2 Conjugation with glucuronic acid
8. Metabolism - Phase II
8.2 Conjugation with glucuronic acid
Deconjugation
8. Metabolism - Phase II
8.3 Conjugation with amino acids
8. Metabolism - Phase II
8.4 Conjugation with glutathione
9. Excretion of chemicals
9.1 By the lungs
-volatile compounds are exhaled by the lungs

9.2 By the liver

-The liver metabolize compounds
Some compounds (e.g. mercury) are however excreted
by bile which contains amphiphathic compounds such
as cholesterol and bile salts
-The liver is the normal excretion way for compounds
with a molecular mass of more than 500 g/mol in
humans (lower for rats)
-Compounds excreted by the liver can be uptaken by
the gastro-intestinal tract....
9. Excretion of chemicals
The kidneys is the most important excretion way

Some animals concentrate the urine (e.g. rats)

9. Excretion of chemicals

Milk is the major excretion way for DDT

in breast-feeding women!

Cl
Cl Cl

Cl Cl
10.Electrophiles

• Why do we cry when we chop onions?

O COOH O S
S S
S
NH2
10.Electrophiles
O
O
S

S O
C
N
10.Electrophiles

• Electrophiles are very reactive and can

destroy DNA

O S
O C
N

small carbonyl- isothiocyanates

rings groups
10.Electrophiles

O
10.Electrophiles

• Electrophiles are reactive

• Defence system: Glutathione

SH
O O O
H
N
HO N OH
H
NH2 O
10.Electrophiles

• Electrophiles are reactive

• Warning system: TRPA1
10.Electrophiles

• Carbonyl compounds
• Teargas O
-developed to be irritating but Cl
rather safe

kloracetofenon, CN
(0.3-1.5 mg/m3)
10.Electrophiles

• Small rings
• Mustard gas
very strong electrophile that affect the skin, eyes
and the lungs

S S
Cl Cl Cl
senapsgas episulfoniumjon
10.Electrophiles

• Mustard gas
-0,02 mg can give blisters
-severe damage on skin
-humidity give worse effects
-infections
 10.Electrophiles
Cl Cl •mustard gas, chemical warfare
S

Cl Cl •nitrogen mustard, chemical warfare

N
•tested as chemotherapy
•too fast hydrolysis
Cl Cl
N •tested as chemotherapy
•good hydrolysis
•not soluble

Cl Cl
N •used as chemoterapy
•chloroambucil

HOOC
11. Specific examples
11.1 Hexane and heptane
11. Specific examples
11.2 Benzene
11. Specific examples
11.2 Benzene
11. Specific examples
11.3 Hydroquinones
11. Specific examples
11.4 Toluene
11. Specific examples
11.5 Naphthalenes
11. Specific examples
11.6 Styrene

good substrate for epoxide hydrolase

11. Specific examples
11.7 PAH (polycyclic aromatic
hydrocarbons)

not a good substrate for epoxide hydrolase

11. Specific examples
11.8 Dichloromethane
11. Specific examples
11.8 Trichloroethylene
11. Specific examples
11.8 Trichloroethylene
11. Specific examples
11.9 Urethane
11. Specific examples
11.10 Acetonitrile
11. Specific examples
11.11 Aromatic amines
11. Specific examples
11.11 Aromatic amines

A-list! B-list!
 11. Specific examples
11.12 Paracetamol

- limited amount of PAPS available for conjugation

-more than a few grams a of paracetamol each day can
give liver damage
11. Specific examples
11.13 N,N-dimethyl formamide
11. Specific examples
11.14 Dimethyl sulfoxide
11. Specific examples
11.15 Acidosis
pH of the blood must be regulated to 7.35-7.40
pH <7 or >8 is lethal
the blood is buffered and we can excrete CO2 in the lungs
and acids with the urine
11. Specific examples
11.15 Acidosis

pKa = 3.8

pKa = 4.8

pKa = 1.2
11. Specific examples
11.15 Haemoglobin
11. Specific examples
11.16 Fluoroacetic acid
11. Specific examples
11.17 HCN