CHAPTER

Factors affecting toxicity

CHAPTER OUTLINE
4
4.1 Introduction ....................................................................................................... 23
4.2 Factors Affecting Toxicity ................................................................................... 23
4.2.1 Host Factors ....................................................................................24
4.2.2 Factors Related to the Toxicant or Associated With Xenobiotics ............26
4.2.3 Environmental Conditions .................................................................28

4.1 INTRODUCTION
It is well known that the sensitivity to the adverse effects of xenobiotics is not
the same for all species. For example, unlike most mammals, rabbits may ingest
large amounts of belladonna (Atropa belladonna), an extremely toxic chemical.
It is also well known that the exposure of a population to a given amount of a
toxicant may not affect each individual to a similar extent because of the occur-
rence of several factors capable of modulating the overall toxic response. Age is
also an important determinant of chemical toxicity for most compounds.
Toxicity is generally higher in younger individuals for a number of reasons,
including, among others, a limited ability to biotransform and excrete xenobio-
tics. Thus, several factors, both of environmental origin as well as pertaining to
individuals or mediated by the xenobiotic itself, may influence the response to
foreign compounds.

4.2 FACTORS AFFECTING TOXICITY

These factors capable of modulating chemical toxicity can be grouped as:
1. Host factors (factors related to subject)
2. Factors related to toxicant or associated with xenobiotics
3. Environmental conditions
Thus, there could be individual or nonindividual factors that can affect the
response of any drug or chemical.

Fundamentals of Toxicology. DOI: http://dx.doi.org/10.1016/B978-0-12-805426-0.00004-4

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23
24 CHAPTER 4 Factors affecting toxicity

4.2.1 HOST FACTORS

Size Size or weight of an individual is an important variable that determines the
dose of a chemical agent required to elicit a given response. If the same amount
of a chemical is given to individuals of different sizes, then the concentration of
the chemical attained in the tissues will be different and, hence, the effect induced
will vary. Large individuals can tolerate a larger dose than individuals of small
size. For this reason, the dosage or the amount of a chemical to be given or the
amount of chemical allowed as acceptable daily intake (ADI) is expressed as per
unit of body weight. The metabolism and activity are proportional to the surface
area of the body and, accordingly, this function has been used for fixing tolerance
limits and for determining the ADI for various chemical agents.
Age Young animals or human infants are uniquely susceptible to chemicals that
are relatively safer during a later period of life. The difference in response during
early life is a consequence of the relative inefficiency of various metabolic and
excretory pathways, the greater susceptibility of certain tissues, immaturity of the
blood brain barrier, and other factors. It is well known that a number of enzyme
systems are poorly developed at birth. This deficiency is primarily quantitative,
with the processes for biotransformation or excretion of the chemical agent being
present but functioning at a level significantly below that of the adult. The oxida-
tive pathways concerned with metabolism of a variety of chemical agents operate
in the newborn at levels well below those of the adults. For example, deficiency
of glucuronyl transferase activity results in enhanced toxicity of chemicals that
are dependent on this route of detoxification and contributes to the accumulation
of unbound bilirubin, which may lead to jaundice. Immaturity of kidney, brain, or
other organs may also render them more sensitive to drugs or chemicals.
Immaturity of the blood brain barrier may allow easier access of the chemical
agent of the central nervous system (CNS) than in the adult.
Species and strains Species variation can greatly influence the toxicity of a spe-
cific compound. The relative toxicity of some chemicals, such as endosulfan,
when tested in several species varied markedly. Often, differences in toxicity are
due to different types of biotransformation, which can take place in various tis-
sues. The compound administered to one species may be rapidly metabolized to a
nontoxic compound or, at times, to a more toxic compound; in a second species,
the same compound may be metabolized at a very slow rate or, in fact, may be
metabolized by another mechanism producing a different metabolite.
Differences in the strain of animals also induce a variation in response of
chemical agents, and such differences have been detected in acute toxicity
measurement of various inbred strains of mice. Because humans are a remarkably
heterogeneous species, the rate of metabolism of any compound may differ
greatly from person to person.
Sex The sex of an animal often has an influence on the toxicity of a chemical
agent. Major differences are shown to be under direct endocrine influence. The
nephrotoxic effect of chloroform in the male mouse can be reduced by estrogen
 4.2 Factors Affecting Toxicity 25

treatment and castration; androgen treatment in the female mouse induces suscep-
tibility to this effect. In such instances, the effect of sex hormones on the
enzymatic biotransformation of a compound might be responsible for the sex
differences. Females generally require a lower dosage than males because of their
smaller size. Because variation in toxicity due to sex is well known, the chemical
agents or drugs must be used with special care during pregnancy because they
could lead to teratogenic effects in females. During lactation, it is important to
remember that some chemicals or drugs may be excreted in milk and may even
act on the offspring. Thus, it is desirable to measure acute toxicity in both male
and female animals of any species.
Feed and feeding The composition of the feed or food can affect the results of
toxicity tests. High-fat diets can sensitize animals to the hepatotoxic effects of
chloroform, whereas high-carbohydrate and high-protein diets provide protection
from these effects. It may be of significance to mention that the activity of hepatic
drug-metabolizing enzyme systems decreases in mice fed a low-protein diet, and
vitamin C deficiency has been shown to affect the rate of drug metabolism in
guinea pigs. Altered reactions have been reported in acute toxicity in animals fed
synthetic diets. Even some food ingredients might induce a toxic reaction during a
drug treatment. For example, sympathomimetic effects due to the interaction of
monoamine oxidase inhibitors and cheese with a high tyramine content have been
demonstrated. These findings indicate that it is necessary to provide a standard
and nutritionally adequate diet in toxicity studies. However, the study of toxicity
in animals with some nutritional deficiency (eg, protein) is of considerable inter-
est. For example, decrease in absorption of the compounds in animals feed prior
to intubation is frequently encountered in oral toxicity tests. Therefore, to assume
more uniform absorption, it is customary to withhold the feed for 16 18 h prior
to intubation. However, starvation-induced catabolic effects may also influence
the results of toxicity. For practical reasons, overnight starvation is a common
practice. Water is offered ad libitum before and after dosage.
Changes in the internal environment Several physiological factors, such as physi-
cal activity, stress conditions, hormonal state of animals, and degenerative
changes in internal organs, are known to influence the toxicity of any compound;
therefore, toxicity tests are performed with healthy animals. Changes in metabo-
lism or body composition during pregnancy and lactation, as well as after major
surgery, could influence toxicological responses.
Alteration of chemical toxicity induced by a compound given prior to the one
undergoing investigation is of great importance. For example, some compounds
may induce increased synthesis of liver microsomal enzymes and influence the
metabolism of another. Inhibition of drug or chemical agent metabolism, displace-
ment of protein binding of a chemical, or inhibition of its renal clearance can also
be accomplished by chemical agents.
Pathologic conditions often modify the effect of chemical agents to a very
considerable extent because animals or humans will respond in different ways to
26 CHAPTER 4 Factors affecting toxicity

the same chemical when they are healthy as opposed to when they are ill. Any
one of these factors will greatly enhance chemical or drug toxicity.
Habitually used drugs The habitual use of certain psychoactive drugs by humans
is known to augment or decrease toxic reactions to drugs in humans. The wide-
spread use of caffeine as tea, coffee, and caffeine-containing soft drinks, alcohol,
and nicotine are commonly abused drugs for social purposes. The habitual, and
particularly excessive, use of these chemicals could affect the sensitivity of
humans to toxic doses of drugs and other chemicals.
Idiosyncratic reaction/toxicity Occasionally, toxicity peculiar to an individual or
that appears in a few persons but not in the general population has been observed.
The incidence of idiosyncratic toxicity is low, varying from less than 1 person in
100,000 to as high as 1 in 10. Idiosyncratic drug reactions often occur after sensi-
tization followed by re-exposure to a drug. A delay of 3 to 4 weeks after a 1- to
2-week course of medication can be seen for some drugs (eg, amoxicillin-
clavulanic acid) before clinical signs become evident, but onset is expedited with
rechallenge. Various drugs are believed to cause immune-mediated idiosyncratic
reactions in humans, including halothane, diclofenac, phenytoin, and sulfona-
mides. Skin reactions to drugs or chemicals taken systemically (reactions such as
urticaria and angioneurotic edema, bronchial asthma, and anaphylactic shock)
have been commonly observed.
Patients with a deficiency of glucose-6-phosphate dehydrogenase, for example,
develop hemolysis after ingesting certain drugs or foods. Some idiosyncratic drug
reactions can be seen after very long latency (up to 12 months) but are usually
not associated with features of hypersensitivity and have been variable in
response to a rechallenge. These are classified as nonimmune-mediated idiosyn-
cratic reactions. Examples of drugs that are known to cause this type of idiosyn-
cratic reaction include troglitazone, valproate, amiodarone, ketoconazole,
disulfiram, and isoniazid. However, some involvement of allergic mechanism can-
not be ruled out.

4.2.2 FACTORS RELATED TO THE TOXICANT OR ASSOCIATED

WITH XENOBIOTICS
Physical state and chemical properties of the toxicant The physical state and
chemical properties of the toxicant such as: (1) solubility in water; (2) solubility
in vegetable oils; (3) the suspending medium; (4) the chemical stability of the
chemical agent; (5) the particle size; (6) rates of disintegration of formulations of
chemicals; (7) the crystal form; and (8) the grittiness of inert substances given in
bulk amounts.
For example, fine particles are more readily absorbed than coarse ones (in the
case of poisons bearing irritating properties, eg, α-naphthylthiourea, zinc phosphide).
Small particles come into contact with a wider surface of the gastric mucosa and
therefore may be more likely to elicit protective vomiting. Some chemicals such as
 4.2 Factors Affecting Toxicity 27

trichlorphon, which is sold as a wettable powder to control external parasites, may

be easily converted in alkaline solutions to dichlorvos, which has an eightfold lower
oral LD50 in rats compared to the parent compound.
Solvents and other substances included in commercial preparations may also
affect the overall toxicity of the active principle(s). Nonpolar solvents may con-
siderably increase the absorption rate of lipophilic poisons, especially when
considering exposure by the dermal route.
Routes and rate of administration Generally, toxicity is the highest by the route
that carries the compound to the blood stream most rapidly. For most xenobiotics,
parenteral routes of exposure entail more prompt and complete bioavailability
than the oral route, often resulting in a lower LD50. The intravenous toxic dose is
greatly influenced by the rate of injection. The gastrointestinal absorption of a
compound varies widely. The differences between oral and parenteral LD50 give
some indication regarding the extent of absorption of a compound. In addition,
orally ingested poisons may undergo first-pass metabolism that, in certain cases,
may lead to almost complete detoxification. As an example, pyrethrum-based
insecticides are practically nontoxic in mammalian species, in which they are
extensively hydrolyzed in the gut, but not in fish, in which a significant amount
of molecules may enter the body through the gills.
Previous or coincident exposure to other chemicals (drug drug Interactions) A
variety of chemicals (drugs, plant toxins, pesticides, environmental pollutants) are
capable of increasing (enzyme inducers) or decreasing (enzyme inhibitors) the
expression and activity of hepatic and extrahepatic phase I and phase II enzyme
systems participating in the biotransformation reactions, thereby modulating the
toxicity of several xenobiotics. Thus, administration of two or more chemicals of
different structures when administered simultaneously may lead to an additive
effect, summation, negative summation, antagonism, or potentiation. If the agents
act on the same organ or tissue and the effect produced is the algebraic sum of their
independent actions, then it is called an additive effect.
If the compound leads to enzyme induction, then this phenomenon takes place
generally slowly because it requires prolonged exposure to the inducing molecule(s).
For a given toxicant, the modulation of the biotransformation capacity mentioned
may have different clinically relevant outcomes according to the nature of the
toxicant under consideration and the nature of its resulting metabolite(s). In the case
of enzyme induction, a decrease in the overall toxicity is expected if the parent com-
pound itself is responsible for the toxic action(s); however, the opposite holds true
whether one or more (re)active metabolites arising from biotransformation reactions
mediates the toxic damage.
On the contrary, enzyme inhibition is expected to entail beneficial effects in
poisonings from toxicants that necessitate a metabolic activation; in certain cases,
inhibitors may be used for therapeutic purposes. Cimetidine is recommended to
treat acetaminophen toxicosis in dogs and cats because of its inhibitory effects on
CYP-mediated generation of the reactive metabolite NAPQI, which is responsible
for severe liver injury occurring in poisoned animals.
28 CHAPTER 4 Factors affecting toxicity

Enzyme inhibition may enhance the toxic potency of chemicals acting per se
(ie, not requiring a metabolic activation) and provide a rationale for explaining
many drug drug interactions.
Drug drug interactions may also occur in the bloodstream. Anticoagulant
rodenticides like warfarin, bromadiolone, or brodifacoum are characterized by rel-
atively long plasma half-lives due to a high degree of plasma protein-binding.
Finally, the co-administration of certain drugs may also modulate the toxicity
of several poisons by acting at the receptor level. For instance, drugs that have
neuromuscular blocking properties that are elicited through their action on nico-
tinic receptors (eg, d-tubocurarine or succinylcholine) may enhance the toxicity of
organophosphates or carbamates, whereas the reverse holds true in the case of
exposure to parasympatholytic agents like atropine, which is widely used as an
antagonist in clinical practice.
Tolerance It is well known that the toxic reaction of an animal to a given dose
of a drug may decrease, remain unchanged, or increase on subsequent adminis-
tration of that dose. A decrease in toxic response is usually called tolerance,
and an increase is called hypersusceptibility. Enzyme induction, or the increased
activity of enzymes concerned with detoxification and elimination of a drug, is
a common mechanism for the development of tolerance to a drug on repeated
administration.
A decrease in the sensitivity of the end-organs to the toxic effects of the drug is
also known to cause tolerance. Chloropromazin, for example, decreases the CNS of
normal albino rats and lessens their locomotor activity. On repeated administration
of chlorpromazine, there occurs an increase in excitatory feedback to the centers
depressed by chlorpromazine with a reluctant decrease in their sensitivity to depres-
sion by the drug. On abrupt withdrawal of the drug, excitatory feedback is no lon-
ger balanced by the depressant action of the drug and a marked increase in activity
of the brain follows, with insomnia and augmented locomotor activity.

4.2.3 ENVIRONMENTAL CONDITIONS

The environment can affect the toxic response to chemicals given to animals or
humans. There are three basic factors in the environment of laboratory animals
used in toxicity testing:
1. The presence of other species of animals, usually humans. Laboratory animals
develop an attitude, maybe hostility, indifference, or affection, depending
largely on the operator in the animal. The attitude of the animal can influence
the toxic response to drugs.
2. The presence of other animals of the same species. The presence of another
male or female of the same species can affect a toxic reaction to certain
chemicals, probably due to aggression or fear.
3. Physical environment. The physical environment under which a toxicity test is
performed causes considerable significance in the toxic response (eg, light,
 4.2 Factors Affecting Toxicity 29

temperature, relative humidity, etc.). Similarly, temperature can influence the

LD50 of several chemicals in rats (change in LD50 values at 37 C and 26 C).
A warm, humid environment is known to enhance dermal absorption as well
as affect toxicity in certain inhalation toxicity studies. Even the light and dark
cycles of the day, number of animals in the cage, construction of the cage,
and seasonal variations are reported to influence the toxicity. Because it is
rarely possible to study the effects of all variations in environmental
conditions, it is important to state under what conditions an experiment was
performed so that one can explain the reason for the variation when
duplicating the experiment and getting divergent results.
For example, high ambient temperatures are reported to enhance the toxicity
of chlorophenols and nitrophenols that cause increased production of heat by
uncoupling mitochondrial oxidative phosphorylation. Conversely, cold tempera-
tures are predisposing factors for α-chloralose, a rodenticide/avicide formerly
used as an anesthetic agent that may induce life-threatening hypothermia, espe-
cially in cats, by acting on hypothalamic thermoreceptors.
Thus, the exposure to poisons may elicit different outcomes according to a
number of factors. In most cases, the variation in the toxic response is based on
the different expressions of enzymes or proteins involved in the kinetics of poi-
sons, which may vary according to diet, species, breed, and physiopathological
factors, or the previous or concomitant exposure to several foreign compounds.
Although until recently such events could not be explained, the current unprece-
dented development of molecular techniques has made it possible to gain insight
into many of the mechanisms underlying most of the differences in the response
to foreign compounds.