Review article ISSN 2321-0125

www.jpbs-online.com
The importance of toxicity testing
David Arome* and Enegide Chinedu
Department of Science Laboratory Technology (Physiology and Pharmacology Technology), University of Jos, Nigeria.
Received on 20 Nov 2013, Accepted on 23 Dec 2013, Available online from 20 Jan 2014

Abstract
Toxicity testing is paramount in the screening of newly developed drugs before it can be used on humans. The essence of
toxicity testing is not just to check how safe a test substance is; but to characterize the possible toxic effects it can
produce. The guiding principles of toxicity testing is to check the effect of the test substances on laboratory animals and
its direct toxic effect on human and secondly, the exposure of laboratory animals to high doses in order to evaluate its
possible hazard on human that are exposed to much lower dose. The present article seeks to highlight the importance of
toxicity testing in the development of therapeutic agents. Toxicity testing is of the following types: acute toxicity studies,
sub-acute toxicity studies and chronic toxicity studies. Toxicity testing employed wide range of test in different species of
animals with long term administration of drug, regular monitoring of physiological, biochemical abnormalities and
detailed post mortem examination at the end of the trial to detect gross or histological abnormalities. Toxic effect of drug
can range from negligible to severe as to preclude further development of the compound. The use of animal in toxicity
testing is most likely to continue for the foreseeable future because of the benefits they offer in examining a whole
functioning organism.
Keywords: Laboratory animals, test substance, toxic effect, toxicity testing

INTRODUCTION Toxicity testing employed wide range of test in different

Toxicity testing is paramount in the screening of newly species of animals with long term administration of drug,
developed drugs before it can be used on humans. regular monitoring of physiological, biochemical
Toxicity testing is the determination of potential hazards abnormalities and detailed post mortem examination at
a test substance may likely produced and the the end of the trial to detect gross or histological
characterization of its action, most of the toxicity testing abnormalities[3]. Dose above therapeutic range are
is carried out on experimental animals[1]. The used in toxicity testing to ascertain the toxic signs of
advantages of using animal models in toxicity testing are action of the drug[2]. The present article seeks to
enormous. These advantages include the possibility of highlight the importance of toxicity testing in the
clearly defined genetic constitution and their amenity to development of therapeutic agents. Toxic effect of drug
controlled exposure, controlled duration of exposure, can range from negligible to severe as to preclude
and the possibility of detailed examination of all tissues further development of the compound. All toxicity study
following necropsy[1]. The information obtained can is supported by; clinical analysis, autophic analysis,
serves as the basis for hazard classification and labeling haematological and haematochemical analysis,
of chemicals in commerce[1]. The essence of toxicity histopathological analysis and statistical presentation
testing is not just to check how safe a test substance is; and data interpretation.
but to characterize the possible toxic effects it can
Importance of toxicity studies
produce. Toxicity testing was given much attention
To establish a dose response curve.
following early 1960s thalidomide catastrophe; with
To ensure safety of new chemicals for use as
thousands of children born worldwide with severe birth
pesticides, drugs, or food additives before they are
defects2. After this incidence many countries of the
registered for general use in industry or doctors clinics.
world have resolved to go for toxicity testing and
To establish the mode of action or mechanism for a
teratogenicity in both sexes so as to prevent further
toxic effect that may have been seen in other studies.
tragedies.

*Corresponding author E - mail: davearome@gmail.com J. Pharm. BioSci. 4(2013) 146-148

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To produce epidemiological studies to explain Arithmetical method of Reed and Muench.

observations in the population, for instance, the long Arithmetical method of Karbar.
investigation into the association of smoking with lung Lorkes method.
To validate new methods of testing or investigation,
Graphical method of Miller and Tainter
particularly those conducted in vitro rather than in
This method is used in the calculation of any ED50 values.
animals[4].
This involves the administration of same volume of
The two basic principles guiding toxicity test in animals different doses of the test substance to various groups.
To check the effect of the test substances on The animals are divided into five groups of ten animals
laboratory animals and its direct toxic effect on per group. Group one animals receive the vehicle to
human. which the test substance is dissolved while the other
Exposure of laboratory animals to high doses in order groups are given different doses of the test substance. In
to evaluate its possible hazard on human that are this method log doses are plotted on a graph against
exposed to much lower doses[3]. probits of the percentage[2].
Toxicity studies are divided into: Arithmetical method of Reed and Muench
Acute toxicity studies This method is a cumulative analysis of values obtained
This is a short term assessment and evaluation of from the result of the study. It is generally assumed that
potential hazard test substance or consequences of the dead of animals would have been caused by the
single dose of a test substance[5]. Acute toxicity testing administration of higher dose of the test substance. The
may be used in risk assessments of chemicals for cumulative dead and survivors are recorded. The
humans and non-target environmental organisms. Acute percent of survival is calculated and the LD50 is
toxicity study is better described as LD50, which is computed[2].
defined as the dose which kills 50% of animals. LD50 is Karbers Method
used for the estimation of the toxicity of the chemical This method involves the administration of different
agents. Acute toxicity provides guidelines on the dose to doses of test substance to various groups which has five
be use in more prolonged studies and it also provides animals each. The first group of animals receive the
the basis for which other testing program can be design. vehicle in which the test substance is dissolved.
In acute toxicity studies rodent are mostly used because However, other groups receive different doses of the
they are economical and readily available and easy to test substance. The animals in each group receives
handle. This test is carried out in each species of animal specific doses, while increment in dose progresses from
as the same route as intended to be use in treatment[2]. group to group (starting from group 2 which receives the
Importance of acute toxicity testing lowest dose). The interval mean of number of mortality
To identify the target organ of toxicity. recorded in each group and dose difference across the
To provides safety measures and monitoring guild lines groups are key parameters in this method. The lethal
for workers involved in the development and testing of dose is calculated using the arithmetical method of
test substances. Karber which is as follows[7].
To provides information needed for the dose selection LD50= LD100 - ( )
in prolonged toxicity studies. Where LD50 = median lethal dose
To generate data containing the adverse effects of a LD100= least dose required to kill 100%
substance on human, animal health and environment. a= dose differ
To provides the basis for which other testing program b= mean mortality
can be design. n= group population
For academics and regulating purpose; classification,
Lorkes method
labelling and transportation of chemical agents
This test was carried out in two phases.
Methods employed in acute toxicity evaluation Phase I
Graphical method of Miller and Tainter.

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In the first phase, nine mice divided into three groups of toxicity testing. New toxicity model under development
three mice each, are given 10, 100, 1000 mg / kg of the include the slug mucosal assay for irritation, transgenic
test substance. After administration of the test animals, the long-term exposure of hepatocyte cultures
substance, observation is made at regular interval to and tissue slices and further development of methods
check for the onset of adverse effect, time to death or for testing for mechanisms of carcinogenicity. The use of
time to recover. The period of observation in this phase I invertebrates, as with the slug for the assessment of eye
is 24 hours. irritation and novel vertebrates such as zebra fish. Stem
Phase II cells remain the great white hope of toxicity testing;
This phase involves the use of three animals divided into their promise remains just that, at the moment, but may
three groups. In this phase, the dose level is either step yet blossom[4].
up or down depending on the outcome of the result
CONCLUSION
obtained from phase I. The animals are administered
Toxicity testing plays a crucial role in ascertaining the
higher dose of 1600, 2900 and 5000 mg/kg. Toxic
toxic effect and characterization of test substance.
symptoms are observed for 24 hours as well as delayed
Toxicity obtained in animal studies occurs with similar
toxic symptoms for 7-14 days. The lethal dose is
incidence and severity in human. The use of animal in
calculated by the formula[6].
toxicity testing is most likely to continue for the
LD50= LD50= (D0D100)
foreseeable future because of the benefits they offer in
D0= highest dose that gave no mortality
examining a whole functioning organism.
D100= lowest dose that produced mortality
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Arome et al., J. Pharm. BioSci. 4(2013)146-148