TOXICOLOGICAL

STABILITY OF
DRUGS
De Jesus, Adia Cavrinni G.
Manalo, Alyssa A.

TOXICOLOGICAL STABILITY
 • To determine maximum expiration
date/shelf life
• To provide better safety to the patients

POINTS OF • To prevent the drug product from

different kind of instability
DISCUSSION • To gather information during pre-
formulation stage to produce a stable
product
 TOXICOLOGY TERMS AND DEFINITIONS
TOXIC - this term relates to poisonous or deadly effects on the body by inhalation
(breathing), ingestion (eating), or absorption, or by direct contact with a chemical.
TOXICITY- describes the degree to which a substance is poisonous or can cause
injury. The toxicity depends on a variety of factors: dose, duration and route of
exposure, shape and structure of the chemical itself, and individual human factors.
TOXIN- a poison of natural origin
TOXICANT- (poison) any chemical that can injure or kill humans, animals, or plants;
a poison. The term “toxicant” is used when talking about toxic substances that are
produced by or are a by-product of human-made activities.
TOXIC SYMPTOM- any feeling or sign indicating the presence of a poison in the
system.
TOXIC EFFECTS- refers to the health effects that occur due to exposure to a toxic
substance; also known as a poisonous effect on the body.
 TOXICOLOGY TERMS AND DEFINITIONS

XENOBIOTICS- a term for “foreign substances”, that is, foreign to the organism. Its
opposite is endogenous compounds. Xenobiotics include drugs, industrial
chemicals, naturally occurring poisons and environmental
HAZARD- potential for the toxicity to be realized in a specific setting or situation.
RISK- defined as the expected frequency of the occurrence of an undesirable effect
arising from exposure to a chemical or physical agent.
ADVERSE EFFECTS- any change from an organism’s normal state
- dependent upon the concentration of active compound at the
target site for a sufficient time
EXPOSURE- the actual contact of the chemical substance with the biological
organism. (It means contact with a hazard.)
DOSE- total amount of a chemical substance administered to an organism
 TOXICOLOGY TERMS AND DEFINITIONS
IMMEDIATE TOXICITY- develops rapidly after a single administration of a
substance
DELAYED TOXICITY- toxic effects occur after the lapse of some period of
time.
TOXICOKINETIC- the quantitation of the time course of toxicants in the
body during the processes of absorption, distribution, biotransformation,
and excretion or clearance of toxicants.
TOXICODYNAMIC- refers to the molecular, biochemical, and physiological
effects of toxicants or their metabolites in biological systems
RESPONSE- The degree of responses depend upon the dose and the
organism
 DRUG STABILITY
USP define stability of pharmaceutical product as “extent to
which a product retains within specified limits throughout
its period of storage and use (i.e. shelf life).
The capacity or the capability of a particular formulation in a
specific container to remain within particular chemical,
microbiological, therapeutically, and toxicological
specifications.

Drug Instability: The incapacity or incapability of a

particular formulation in a specific container to remain
within a particular chemical, microbiological,
therapeutically, physical & toxicological specification.
 POTENTIAL INSTABILITY ISSUES OF FULL PACKAGED
PRODUCT (FPP)

• Loss/ increase in concentration of API

• Formation of (toxic) degradation products
• Modification of any attribute of functional relevance
• Alteration of dissolution time/profile or bioavailability
• Decline of microbiological status
• Loss of package integrity
• Reduction of label quality
• Loss of pharmaceutical elegance and patient
acceptability
 STEPS
IN
TOXICITY
TESTING
 DIFFERENT AREAS OF TOXICOLOGY
• Analytical Toxicology
• Applied Toxicology
• Clinical Toxicology
• Environmental Toxicology
• Forensic Toxicology
• Industrial Toxicology
• Occupational Toxicology
• Veterinary Toxicology
• Food Toxicology
• Regulatory Toxicology
OPENING
 TEST REPORT

The test report should include the following

information:
• Test substance
• Physical nature
• Purity
• Physicochemical properties
• Identification data
• Source of substance
• Batch number
 NECESSITIES OF TOXICOLOGICAL STUDIES

• Benefits-risk ratio can be calculated

• Prediction of therapeutic index
𝑀𝑎𝑥𝑖𝑚𝑢𝑚 𝑡𝑜𝑙𝑒𝑟𝑎𝑡𝑒𝑑 𝑑𝑜𝑠𝑒
Therapeutic index=
𝑀𝑎𝑥𝑖𝑚𝑢𝑚 𝑐𝑢𝑟𝑎𝑡𝑖𝑣𝑒 𝑑𝑜𝑠𝑒
• Smaller ratio, better safety of the drug
 SOURCES OF TOXIC SUBSTANCE

Classified based on their:

• Chemical nature
• Mode of action
• Class (exposure class and use class)
• Exposure class: food, air, water, soil
• Use class: drug as drug of abuse, therapeutic
drugs, agriculture chemicals, food additives,
pesticides, plant toxins, and cosmetics
 LOCAL ADVERSE EFFECTS

The site of action takes place at the point of

contact.

Sites:
• Skin
• Mucous membrane of the eyes, nose, mouth,
throat or anywhere the along the respiratory
or gastrointestinal system
 SYSTEMIC ADVERSE EFFECTS

Requires absorption and distribution to a distant site.

Most chemical substances can produce systemic effects

Target organs:
- CNS
- Circulatory system
- Blood and hematopoietic system
- Liver or GI
- Kidney
- Lung
 CUMULATIVE EFFECTS

• Over a period of time, the material is only

partially excreted and the remaining quantities
are gradually collected.
• The retained toxic compound accumulates and
becomes great enough to cause adverse
effect.
 VARIOUS INTERACTIONS OF CHEMICAL SUBSTANCES

ADDITIVE- Combined effect of two chemicals is equal to the sum of

the effects of each (2+3=5)
SYNERGISTIC- Combined effect of two chemicals are much greater
than the sum (2+2=20)
POTENTIATION- One substance does not have a toxic effect on a
certain organ or system but when added to another chemical is
already toxic
ANTAGONISM- Two chemicals interfere with each others in actions
 DURATION AND FREQUENCY OF EXPOSURE

ACUTE- A single exposure lasting less than 24 hours

SUBACUTE- Repeated exposure lasting 1 month or
less.
SUBCHRONIC- Repeated exposure lasting from 1
month to 3 months
CHRONIC- Repeated exposure lasting more than 3
months (over months or years)
 FACTORS THAT INFLUENCE TOXICITY

• Physicochemical composition of the toxicant (solids, liquids, particle size, pH)

• Dose and concentration (aspirin tablets, dilution)
• Routes of administration (inhalation> IV> IP> SC> IM> ID> oral> topical)
• Metabolism of the toxic agent (more polar or more toxic)
• State of health (person with severe hepatic or renal disease, hypertension, head
injuries)
• Age and maturity (gray-baby syndrome, reduced pharmacokinetics in elderly people)
• Nutritional state and dietary factors (stomach contents, types of food, proteins)
• Pharmacogenetics or idiosyncrasy (succinyl choline, aspirin, fava beans)
• Sex (erythromycin, lipid sol., large BV and tissue mass in men which dilute the
chemical)
• Environmental (temperature, occupation (liver metab), living conditions)
• Chemical interactions (the increase or decrease of toxicity by simultaneous or
consecutive exposure to another one)
EVALUATION OF SAFETY OF CHEMICAL AND DRUGS
STABILITY CHAMBER
 COMMONLY USED
EQUIPMENT IN TOXICOLOGY
LABORATORY
 TYPES OF TOXICOLOGICAL STUDIES
• Systemic toxicological studies
• Reproductive toxicological studies
• Local toxicity studies
• Hypersensitivity studies
• Genotoxicity studies
• Carcinogenicity studies
 TYPES OF LOCAL TOXICITY STUDIES
• Dermal toxicity studies
• Dermal photo-toxicity studies
• Vaginal toxicity studies
• Rectal tolerance studies
• Ocular toxicity studies
• Parenteral drugs
• Inhalation toxicity studies
EXPERIMENTAL TOXICITY TESTS
ACUTE TOXICITY STUDIES
MAGNITUDE USE OF ANIMALS
ACUTE TOXICITY TESTS: SKIN AND EYE STUDIES (DERMAL)
ACUTE TOXICTY TESTS: PYRAMIDAL SINGLE DOSE TEST
 SKIN SENSITIZATION TESTS

Skin sensitization tests are carried out using the guinea pig as a model.
Skin sensitization is assessed using the Draize test, open
epicutaneous test, optimization test, split adjuvant test, guinea pig
maximization test (GPMT), Buehler test, and murine local lymph
node assay (LLNA). The LLNA method is used as an alternative to the
guinea pig Draize test, and it is widely accepted that this method
meets regulatory requirements. In the LLNA test, the test substance is
applied on the surface of the ears of a mouse for three consecutive
days, and the proliferation of lymphocytes in the draining lymph node
is measured at the end.
 ALLERGENICITY/ HYPERSENSITIVITY TOXICOLOGY STUDIES

• GUINEA PIG MAXIMIATION

• Determination of maximum non-irritant dose
• Evaluation of erythema and oedema
• LOCAL LYMPH NODE ASSAY
• Mice of one sex (either male or female)
• Drug treatment given on ear skin
• Auricular lymph node dissection after 5 days
• Increase in 3h-thymidine for evaluation
PROLONGED TOXICTY STUDIES
REFERENCES
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3. Klaassen CS. Principle of toxicology and treatment of poisoning. In: Parker BK, Blumenthal D, Buxton L (eds.).
Goodman & Gilman’s; manual of pharmacology & therapeutics. McGraw Hill. 2008: 1115-1119.
4. Woolley A. A guide to practical toxicology, evaluation, prediction and risk. 2nd ed., Informa Health Care. New York,
London. 2008.
5. Monosson E. Toxicity testing methods; Encyclopedia of earth topics. [updated 2013]. Available from:
http://www.eoearth.org/view/article/1566.73/.
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7. Turner R. Acute toxicity: The determination of LD50. Screening Methods In Pharmacology. Academic Press, New
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