CHAPTER 3:

TRANSPORT PROCESS AND

TOXICOLOGY

FOR ECH4106
Learning Objective
To describe relationship between dose, exposure with
toxic response
To determine appropriate technique for toxicity effect
quantification of xenobiotic
 Content:
Toxicity of xenobiotic
Basic Toxicology
Exposure, Dose Response
Toxicokinetics

toxic
therapeutic effect
effect increasing dose
 How Xenobiotics Cause Toxicity
Some xenobiotics cause toxicity by disrupting normal cell functions:

Bind and damage proteins (structural, enzymes)

Bind and damage DNA (mutations)

Bind and damage lipids

React in the cell with oxygen to form

free radicals which damage lipid, protein,
and DNA
Types of Toxic Effects
Death - arsenic, cyanide

Organ Damage - ozone, lead

Mutagenesis - UV light

Carcinogenesis - benzene, asbestos

Teratogenesis - thalidomide
Toxicology
Definition: the study of the effect of poisons on the
function of living systems
Chemical agents that cause toxicity include:
Drugs
Insecticide/ herbicides
Plant toxins
Animal toxins
Chemical weapons
Radioactive elements
Local vs. Systemic Effect

Systemic: adverse effect at distant Local: adverse effect at point of

location from the route of entry. exposure/entry
eg: arsenic effects to the blood,
nervous system, liver, kidneys
and skin; benzene effects to the
bone marrow
Systemic compound: often have
target organ which they
accumulate
When several substance
exposure: synergetic effects- eg:
smoking+asbestos, alcohol+
chlorinated compounds
BASIC CHEMISTRY
BASIC CHEMISTRY

Dr. NA 08/9
Distribution and storage
Fat-nonpolar compound/lipophilic (eg.PCBs)
Blood plasma-compound bound by blood (eg. mercury
ions)
Bone- lead, radium, fluoride
Kidneys-cadmium
Thyroid gland-iodine (Iodine 131-radioactive, half-life 8days,
absorbed by thyroid gland, as it decays it may destroy
thyroid)

Dr. NA-09
GENERAL HEALTH EFFECTS OF SOLVENT
EXPOSURE

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GENERAL HEALTH EFFECTS OF GASES

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HEALTH EFFECTS TERMINOLOGY

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TOXICOLOGICAL TERMINOLOGY
LD50= dose required to kill
50% test subjects
LC50= concentration
required to kill 50% test
subjects
IDLH= concentration likely
to cause death, immediate
or delayed permanent
health effect if a person
cannot immediately escape,
eg: CO (1200ppm), H2S
(100ppm), phosgene
(2ppm)
 Safety for Workers at Workplace.
(OSHA, NIOSH)
TLV (Threshold Limit Value)= exposure standard & should not
exceed this value, concentration that can be tolerated for
8hr/day, 40hr/wk
IDLH
REL (Recommended Exposure limit)
PEL (Permissible Exposure limit): maximum amount or airborne
concentration of a substance to which a worker may be legally
exposed
SKIN LD50

Dr. NA 08/9
INHALATION LC50

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 Toxicity of Gases

Blister =phosgene exposure)

hyperkeratosis=arsenic poisoning)
 ORGANOPHOSPHATES

GA, GB, GD & VX= all nerve toxin, contain phosphorus

Dr. NA 08/9
ARSENICAL

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CYANOGENS (CN)

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CN Poisoning

In blood, CN bind to
hemoglobin (Hb) of red
blood cell, then taken
up by enzyme
cytochrome oxidase ,
which stop cells from
absorbing O2.
cyanide effectively
chemically asphyxiates
the body.
 SOURCE & TYPE IN
TOXIC WASTEWATER
a) Domestic Wastewater
Contain ammoniacal nitrogen up to 50 mg/l
Septic: may contain sulfide up to 50 mg/l
Damage to aquatic fauna
b) Stormwater
Influenced by nature of drainage area & frequency of storms
Mainly associated with heavy metals

Dr. NA-09
 SOURCE & TYPE IN
TOXIC WASTEWATER
c) Agricultural Wastes
Materials used for fertilizers & pest control
Fertilizers: oxidized nitrogen cause human
toxicological problems
Pesticides & herbicides: aquatic toxins
d) Industrial Wastes
e) Leachates
Leach out when surface or ground water has
access

Dr. NA-09
Factor determining chemical
toxicity effect
1. Exposure
2. Dose
3. Intake route and rate
1) Exposure-Response

Dr. NA-09
 Exposure Period
Acute 1 day
Subacute 10 days
Subchronic- 2weeks to 7yrs
Chronic- 7yrs to lifetime

Dr. NA-09
Example of Exposure

Dr. NA-09
Example of Exposure

Dr. NA-09
 2) DOSE-RESPONSE
Study on adverse effects caused by exposure of living
organisms to chemical substances
Human response to toxic chemicals

H
Death
U
m
a Clinical
n

R Sub-clinical
E
S
Behavioral
P
O
n
S
e

Low <Dose of Toxic Agent> High

Dr. NA-09
 Dose-Response relationships
Dose:
The amount of agent actually deposited within the body
Typically, the distinction between exposure and dose is blurred,
although in reality, significantly different doses can result from the
same exposure

Response:
The biological response to an agent
LD50 = median lethal dose (mg/kg of body weight)
LC50 = median lethal concentration (concentration of
substance /volume of inhaled air)

Dr. NA-09
Distinction between exposure and dose
Exposure is outside the body
Dose is inside the body

Definition of response- Change in structure or function,

morbidity, or mortality
Define and characterize endpoint

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Important Issue
Challenges in obtaining dose-response relationships
Characterization of exposure or dose
Assessment of response
Selection of dose-response model to fit the observed data

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Issues in Understanding Response
Acute versus delayed onset
Latent period confounds many epidemiologic studies

Short-term versus chronic disease

Irreversibility

Spontaneous incidence
Function of age
Tease out agent-produced component from background
Hundreds of causes of nonspecific effects

Dr. NA-09
Dose-Response Models

Dr. NA-09
Dose-Response Curve

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 37

Preliminary toxicity testing

NOAEL (no observed adverse effects level)
Highest concentration that does not a toxic response

LOAEL- lowest observed adverse effects level

Lowest concentration that produces a toxic response

100
Toxic response

75

50
LOAEL
NOAEL 25

0
-9 -8 -7 -6 -5 -4 -3
log [drug] (M)
 38

LD50 values for different toxins

Toxicity rating Example LD50 (mg/kg)

Slightly toxic Ethanol 8000

(5-15 g/kg)
Moderately toxic Sodium chloride 4000
(0.5-5 g/kg) Parathion 1300
Very toxic Aspirin 300
(50-500 mg/kg) Paracetamol 300
Extremely toxic Theophylline 50
(5-50 mg/kg) Diphenhydramine 25
Super Toxic Potassium cynanide 3
(<5 mg/kg) Digoxin 0.2
Tetrodotoxin 0.01
Botulinum toxin 0.00001 (10 ng/kg !)
 39

Therapeutic index
The ratio of the dose of the drug that produces an unwanted
(toxic) effect to that producing a wanted (therapeutic) effect
= LD50 / ED50

Therapeutic window Therapeutic window

response (%)

response (%)

log [drug] (M) log [drug] (M)

Small TI: e.g. warfarin Large TI: e.g. penicillin,
aspirin 39
 Example

This is a figure of two different dose

response curves. You can obtain a
different dose response curve for any
system that the drug effects. When
you vary the drug, this is the
Independent variable, what you are
measuring is the % of individuals
responding to the drug. Here we see
the drugs effects on hypnosis and
death. Notice that the effective dose
for 50 % of the people is 100 mg and
if you double the dose to 200 mg then
1 % of your subjects die. Thus, if you
want to use this drug to hypnotize 99
% of your subjects, in the process
you will kill 2-3 % of your subjects.
 41

Preliminary toxicity testing

The oral LD50 of a new drug was determined in rats.

Q. What can this value tell us:

A. Short term, lethal effects

B. Long-term, lethal effects
C. Long-term, non-lethal effects
D. Potential Type B adverse drug reactions
E. Lethal dosage when injected
F. Toxicity in young and old humans
3) Intake route and rate
Inhalation effect within seconds
Absorption though skin effect within 30min
Ingestion- effect within hours
Routes of Administration
TOXICOKINETIC

3 KINETICS OF TOXICANT
IN HUMAN BODY:
1. ABSORPTION
2. DISTRIBUTION
3. ELIMINATION
 45

Toxicokinetics - the study of the time course of toxicant

absorption, distribution, metabolism, and excretion
How can we predict variability among individuals?
How can we extrapolate from animal models to humans?

Dosage Plasma Site of

Toxic
Exposure Conc. action
Effects

Toxicokinetics Toxicodynamics
 46

Toxicokinetic (TK) processes

ABSORPTION DISTRIBUTION METABOLISM EXCRETION

EXTERNAL BLOOD PLASMA
PHASE-1 KIDNEYS
MEMBRANE
Oxidation LIVER
BARRIERS
xenobiotic lungs
skin TISSUES PHASE-2 saliva
G.I. tract pools sweat
conjugation
lungs depots breast milk
sinks
PHARMACOKINETICS
Refers to rate of body mechanisms deal with
toxicants (absorbed, metabolized, eliminated)
Fast rate: may tolerate toxic dose when received
Slow rate: accumulates to a critical concentration
Many models rely on 0-order, 1st-order, or 2nd-
order reaction kinetics combined with
mathematical approaches

Dr. NA-09
1) Absorption of toxic:
Diffusion across plasma membrane

Passive diffusion- gases, hydrophobic

compound (eg. Benzene), small polar
molecule (eg. alcohol)
Charged compound, hydrophilic
molecule- active/ facilitative transport
 Xenobiotics at Work
TOXICOKINETICS

Xenobiotic

Excretion
 1) Absorption: Fickss law and Diffusion

dD/dt = KA (Co - Ci) / t

Where;
dD/dt = rate of mass transfer across the membrane
K = constant (coefficient of permeability)
A = Cross sectional area of membrane exposed to the
compound
C0 = Concentration of the toxicant outside the membrane
Ci = Concentration of the toxicant inside the membrane
t = Thickness of the membrane
Bioavailability
Definition: the fraction of the administered dose
reaching the systemic circulation
for i.v.: 100%
for non i.v.: ranges from 0 to 100%

e.g. lidocaine bioavailability 35% due to

destruction in gastric acid and liver metabolism

First Pass Effect

 52

FIRST PASS
EFFECT
Intestinal vs. gastric
absorption

Wilkinson, NEJM 2005

 Bioavailability (F)
70

60 i.v. route (AUC)o

50 (AUC)iv
Plasma concentration

40

30 oral route
20

10

0
0 2 4 6 8 10

Time (hours)
Principle of Xenobiotic distribution
For xenobiotics taken by routes other than the iv, the extent of
absorption and the bioavailability must be understood in order to
determine whether a certain exposure dose will induce toxic effects
or no
Distribution is second phase of TK process. This defines where in the
body a xenobiotic will go after absorption
Vol distribution (Vd) =Volume into which a drug appears to
distribute with a concentration equal to its plasma
concentration

Amount of drug in body

Vd =
Concentration in Plasma
 55

Co V = Dose / Co
Ln of
Blood (or
Plasma)
Conc.

Time

Vd can be calculated after an IV dose of a substance that

exhibits "one-compartment model" characteristics.

Vd = Dose / Initial Conc

2) Distribution/Disposition of
Xenobiotics
absorption

distribution

excretion
General Scheme of Xenobiotic Metabolism

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

Phase I Metabolites Phase II Metabolites

(oxidative) (synthetic)
size
Bioactivation polarity ionization
Detoxification
Detoxification functionality water solubility
Increase excretability
Protein Binding
Some xenobiotics has high protein binding

Plasma protein Drug

bound drug Free drug in
tissues

C
Cnormal binding =
P
(1 fu) + fu
fu = fraction unbound Pnormal
Effect of pH on Vd
 Weak Acids and Weak Bases
HA <==> H+ + A- B + H+ <==> BH+
[ UI ] [I] [ UI ] [I]

pKa = pH + log(HA/A-) pKa = pH+ log(BH+/B)

pH = 2 pKa = 4.5 (a weak acid) pH = 7.4

0.1 = [ I ] [ I ] = 9990

100 = [ UI ] [ UI ] = 100

100.1 = total drug = 10090

 Salicylate is a Weak Acid (pKa 3.5)

TISSUES BLOOD URINE

(pH 6.8) (pH 7.4) (pH variable)

SH SH SH

H+ + S-
H+ + S- H+ + S-

Acidosis Alkalosis
Remember Henderson-Hasselbalch?

protonated species
Log = pKa pH
unprotonated species

OR . . .

protonated/unprotonated = 10pKa-pH
Dose was 150 mg IV . . . Vd = ?

Cp at
t=0
~ 7.5 mg/L Vd = dose / Cp
= 150 / 7.5
= 20 L
Half-life: time taken for Cp become half

Half-life
Time taken for the concentration of a chemical in the body
(usually measured in blood) to fall by half
Majority of excretion (~97%) completed within 5
half-lives.
Half life

1 half-life

2 half-lives
K = slope

Slope =
the proportion of
drug elimination
per unit time
(natural log graph)
 67

3.) Elimination of toxic

Zero order: constant rate of elimination
irrespective of plasma concentration.
First order: rate of elimination proportional to
plasma concentration. Constant Fraction of drug
eliminated per unit time.

Rate of elimination = constant (CL) x Conc.

Example: Elimination of Ethanol
Metabolism of ethanol is 0-order reaction (elimination is constant
regardless of conc.) If an average human is able to metabolize 10 ml of
ethanol per hour, how long a time is required to eliminate a 6-pack of
beer containing 5% alcohol? 1 bottle of beer contained 436 ml
Solution: 13.1hr

Dr. NA-09
Eg2: Half-life for elimination of methyl
mercury from human body
Find half-life of methy-Hg in a person surviving poisoning
by a fungicide exposure. The rate of methyl-Hg absorption
is obeying 1st order rate of reaction. The average excretion
rate is 1.75% of total body burden per day.
Solution: or

ln C
C = Coe -kt Co
t =-
k
ln 0.5
t1 = - 1 = 39.6days
2 0.0175 /day
 Plasma Concentration 10000

1000
Zero Order Elimination

100

10

1
0 1 2 3 4 5 6

Time
logCt = logCo - Kel . t
2.303
 71

First Order Elimination

dC/dt = k
Plasma concentration

14
12 -Kel.t
Ct = Co e
10
lnCt = lnCo Kel .t
8
6 logCt = logCo - Kel . t
2.303
4
2
0 y = b a.x
0 5 10 15 20

TIME (hours)
Cp after single iv injection

Plasma Concentration
Distribution and Elimination
10000

Elimination only
1000
C0
100

10
Distribution equilibrium
1
0 1 2 3 4 5 6
Time
 lnCt = lnCo Kel.t

Vd = Dose/C0

When t = 0, C = C0, i.e., the concentration at

time zero when distribution is complete and
elimination has not started yet. Use this value
and the dose to calculate Vd.
 lnCt = lnCo Kel.t

t1/2 = 0.693/Kel

When Ct = C0, then Kel.t = 0.693. This is the

time for the plasma concentration to reach half
the original, i.e., the half-life of elimination.
Two Compartment Model

Assumes xenobiotic
enters the first
compartment
Assumes that xenobiotic
is distributed to the
second compartment and
a pseudoequilibrium is
established
Elimination is from the
first compartment

75
Fate of toxicant in human body
Upon reaching organ, toxicants can cause 3 event (apart
from storage):
1. Biotransformation
2. Elimination
3. Formation of chemical-receptor complex