KINETIC PRINCIPLES

AND
STABILITY TESTING

Theoretic considerations regarding drug degradation

 Drug Stability:
▪ Drug stability means the ability of the pharmaceutical dosage form
to maintain the physical, chemical, therapeutic and microbial
properties during the time of storage and usage by the patient.
▪ It is measured by the rate of changes that take place in the
pharmaceutical dosage forms.
▪ The United States Pharmacopoeia defines the drug stability
as “the extent to which a particular drug product remains
within specified established limits throughout its storage and
use i.e. shelf life”.
Shelf life:
 It is the length of time that a drug product may be stored
without becoming unfit for use.
 It is the time span between the date of manufacture and the expiry
date.
 A drug that has passed its shelf life might still be safe for
consumption, but its quality is no longer guaranteed.
 Up to 10% degradation of a drug product is pharmaceutically
acceptable during shelf life.
Expiry date:
▪ It means the date after which the drug can not be used
because the concentration becomes lower than therapeutic
concentration.
▪ In addition, some products of drug degradation are toxic and
harmful to patients.
▪ Note! After the opening of the drug container, the expiry date will
be shorter because of the effects of external factors.
▪ Example: Eye drops can be used for one month after opening.
 Stability Studies:

 Long-Term (Real-Time) Stability Testing Under Normal Conditions of

Temperature, Humidity and Light.
 Stability Testing Under Accelerated Conditions of Temperature, Humidity and
Light.
 Usual time points for testing are at 0, 3, 6, 9, 12, 18, 24, 36 months, etc.

General case Accelerated Long term

40 oC + 2 oC 25 o C+ 2 oC
75%RH + 5% RH 60 %RH + 5% RH

Drug products Accelerated Long term

intended to be stored 25 o C+ 2 oC 5 oC + 3 oC
in a refrigerator 60%RH + 5% RH
Stability chamber:
 Objectives of Drug Stability Study:
 To determine maximum expiration date/ shelf life.
 To provide better drug safety to the patients.
 To prevent the drug product from different kind of instability causing factors.
 To provide better storage condition.
 To determine the type of packaging components to be used for a particular drug
product.
 To gather information during pre-formulation stage to produce a stable product.
 Types of Stability:
1) Chemical Stability:
Each active ingredient retains its chemical integrity and labeled potency within the specified
limit.

2) Physical Stability:
The physical stability properties including appearance, palatability, uniformity, dissolution,
disintegration, suspendibility etc. are retained

3) Microbiological Stability:
Sterility or resistance to microbial growth is retained according to specified requirements.
4) Therapeutic Stability:
Therapeutic activity remains unchanged.
5) Toxicologic Stability:
No significant increase in toxicity occurs.
Factors affecting drug stability:
a) pH
b) Temperature
c) Ionic strength
d) Acid-base catalysis
e) U.V. light
f) Moisture
g) Oxygen
h) Concentration
i) Drug Incompatibilities
j) Type of dosage form
a)-pH:
▪ Acidic and alkaline pH influence the rate of decomposition of most drugs.
▪ Many drugs are stable between pH 4 and 8.
▪ Weekly acidic and weekly basic drugs show good solubility when they are
ionized and they also decompose faster as they are in charged form. So to
prevent instability, a water miscible solvent is introduced into the product. It
will increase stability by suppressing ionization and reducing the water
activity by reducing the polarity of the solvent.
▪ Reactions catalyzed by pH are monitored by measuring degradation rates
against pH.
▪ Some buffers such as acetate, citrate, lactate, phosphate and ascorbate are
utilized to prevent drastic change in pH.
b)-Temperature:
▪ High temperature accelerates oxidation, reduction and hydrolysis reaction
which lead to drug degradation.
Room temperature  ~ 25-30oC
Cold temperature  ~2-8oC (refrigerator)
Freeze storage  ~ -20 to -10oC
▪ Thermolabile drugs are affected by high temperatures.
▪ Different products need storage at different temperatures.
▪ Examples of products needing storage at low temperatures are vaccines,
sera, biopharmaceuticals, biological fluids, plasma etc.
c)-Ionic strength:
▪ Ionic strength of a medium is related to concentration of ionic species in it.

▪ Addition of electrolyte in formulation changes ionic strength which is important

in investigation of effect of pH on drug degradation.

▪ So, for this case, ionic strength of different buffers used in preparations must
be kept constant to avoid interference with results.
d)-Acid-base catalysis:
▪ Catalyst is an agent that accelerates the rate of a reaction without itself being
consumed.
▪ Hydrolysis is catalyzed by hydrogen ions (as H3O+in solutions) or hydroxyl
ions. So pH is also an important factor.
▪ Specific acid catalysis is catalyzed by hydrogen ions and specific base
catalysis is catalyzed by hydroxyl ions. So, relationship b/w pH and
degradation rate of drug is plotted as graph.
▪ Sometimes besides H3O+ or OH-, other components of formulation act as
catalyst for example phosphate and acetate buffers lead to hydrolysis of
amide bond in chloramphenicol. So, compatibility of active ingredients and
excipients must also be checked to ensure stability.
e)-Light:
▪ It affects drug stability through its thermal effect which leads to oxidation
▪ Shorter the wavelength, more will be the energy, and more will be the
degradation.
UV  200-400 nm    more degradation
Visible  400-800 nm    less degradation
▪ Photolysis may be protected by:
1. Suitable packaging in amber coloured bottles for light sensitive drugs
2. Card board outers
3. Aluminum foil
f)-Moisture:
▪ Water catalyzes chemical reactions as oxidation, hydrolysis and reduction reaction.
▪ Water also promotes microbial growth.
▪ Moisture caused hydrolysis may be prevented by several ways such as

1. storage in air-tight containers,

2. avoiding contact with moisture,
3. use of desiccants etc.
g)-Oxygen:
▪ Exposure of drug formulations to oxygen affects their stability
▪ Oxygen caused oxidation may be prevented by

1. storing the drug product in air-tight container,

2. use of antioxidants, buffers etc.
h)-Concentration:
▪ Concentrated preparations tend to degrade slowly than dilute ones as seen in
case of simple syrup B.P and simple syrup U.S.P. where high sucrose
content prevents microbial growth and thus protects the product.
i)-Drug incompatibility:
▪ Reactions between components of pharmaceutical dosage forms itself or
between formulation and the container material can also cause instability. So,
appropriate containers must be used for preparation and storage.
j)-Type of Pharmaceutical dosage form:
▪ Solid dosage forms are more stable than liquid dosage forms due to absence
of water.
 Drug Instability:
▪ “The incapability of a particular formulation in a specific container to remain within
a particular chemical, microbiological, therapeutic, physical & toxicological
specification”.
Types of drug instability / degradation:

1. Physical degradation
2. Chemical degradation
3. Microbiological degradation
4. Therapeutic degradation
5. Toxicological degradation
 1-Physical Degradation:
▪ In physical degradation, changes in physical nature of the drug product take place.
Types of physical degradation are :
1. Crystal formation in pharmaceutical preparations

2. Loss of volatile substances from pharmaceutical dosage forms

3. Loss of water

4. Absorption of water

5. Change in crystalline form

6. Colour changes
 1-Crystal formation in pharmaceutical preparations:
Causes:

a. Polymorphism phenomena: i.e. Chloramphenicol change of amorphous (active form) to

crystalline form.

b. Saturated solution: by temperature difference, precipitation of solute may occur (less solubility).

c. In suspension: when very fine powder is used, a part of suspending agent will dissolve then precipitate
as crystal as in ophthalmic preparations.

Prevention:

▪ For solutions
Stabilizers are added

▪ For suspensions
· Minimum temperature fluctuation should be managed
· Incorporation of surface active agent
· By increasing viscosity of suspending material
 2-Loss of volatile substances from pharmaceutical
dosage forms:
▪ Volatile components such as alcohol, ether, Iodine, volatile oils, camphor, menthol etc. escape
from the formulations.
▪ Examples of such dosage forms are:

a. Aromatic waters
b. Elixirs
c. Spirits
d. Some types of tablets which contain aromatic water (Nitroglycerin tablets)
Prevention:
▪ Such products should be placed in a well closed container
▪ Stored at low temperature
 3-Loss of water:
▪ This tendency depends on temperature and humidity of surrounding environment.

▪ This can be seen in the following dosage forms:

a. Saturated solution: by loss of water they become supersaturated and precipitated as crystals

b. Emulsions: Loss of water leads to separation of the two phases and change to other type (o/w may
become w/o emulsion)

c. Creams: especially oil/water, they become dry by loss of water

d. Pastes

e. Ointments: especially aqueous base ointments

Prevention:
▪ Humectants are added to dosage forms which are defined as hydrophilic substances added to aqueous
phase to absorb water from atmosphere and prevent its loss from the dosage forms. Example: Glycerin
▪ Products should be placed in well-closed containers

▪ Use glass containers

 4-Absorption of water:
▪ Hygroscopic drugs absorb the water from external atmosphere causing the physical degradation.
▪ Absorption of water depends on temperature and humidity of surrounding environment.
▪ This phenomena can be seen in the following pharmaceutical forms:

a. Powders: Liquefaction and degradation may occur as a result of absorption of water

b. Suppositories (in which base made from hydrophilic substances, Glycerin, Gelatin, polyethylene
glycol, is used), consistency becomes jelly-like in appearance.
c. Gelatin capsule shells: may distort or dissolve.
Prevention:
▪ Products should be placed in well-closed containers
▪ Use moisture adsorbent (silica gel)
 5-Change in crystalline form:
▪ Crystalline form of components is sometimes changed.
▪ For Example: Cocoa butter which is capable of existing in four polymorphic forms. When
temperature changes, stable form is converted to other forms which are unsuitable for use.
▪ It is important that formulated products should contain a stable crystalline form of the drug.
 6-Colour changes:
▪ Colour changes are of two types.

1. Loss of colour due to pH change or presence of reducing agent

2. Development of colour due to exposure to light

Prevention:
▪ pH should not be changed
▪ Exposure to light should be avoided
▪ UV light absorbing material should be incorporated.
 2-Chemical degradation /instability:

▪ Chemical degradation of a dosage form leads to lowering of therapeutic

agent in the dosage form, formation of toxic product, decreased
bioavailability etc.
Types: It occurs through several pathways such as
1. Hydrolysis,
2. Oxidation- reduction reactions,
3. Complex chemical reactions
4. Photolysis etc.
 1-Hydrolysis:
▪ “Hydrolysis is a reaction involving the breaking of a bond in a molecule using water”.
Examples of hydrolysis in drugs:
Esters  Aspirin, Nitroglycerin
Amides  Indomethacin, Lidocaine, Chloramphenicol, Paracetamol
Lactones  Pilocarpine, Spironolactone
Factors affecting Hydrolysis:

▪ Moisture
▪ pH
▪ Temperature
▪ Type of the solvent
Protection against Hydrolysis:
 Adjustment of pH
 Avoiding contact with moisture (Hydrolysis susceptible drugs such as penicillin and
derivatives can be formulated in the dry powder form for reconstitution instead of a
liquid solutions or suspensions).
 Use non-aqueous solvent for preparations
 Pack in air-tight containers.
 Use desiccants.
 In liquid dosage forms, hydrolysis is acid or base catalyzed; therefore, use an
appropriate buffer to maintain pH.
 Hydrolysis of certain drugs such as benzocaine and procaine can be decreased by
the addition of specific chelating agent like EDTA to the drug solutions.
 2-Oxidation-reduction reactions:
▪ It the process of Addition of oxygen, Removal of hydrogen, Removal of electrons

Types:
Oxidation has two types

1-Auto-oxidation “Oxidation in which the oxygen present in the air is involved”. This process
proceeds slowly under the influence of atmospheric oxygen e.g. Oil, fats & unsaturated
compound can undergo auto-oxidation
2-Photo-oxidation “Oxidation in which instead of oxygen, light is involved.”
This type is less frequently encountered e.g. It occurs in adrenaline, riboflavin & ascorbic acid
etc.

Examples of drugs that may undergo oxidation are Ibuprofen, Codeine, Morphine, Cimetidine,
Diazepam etc.
Protection against Oxidation:
 Avoiding contact with air.
 Pack in air-tight containers.
 Use of reducing agents (Sodium metabisulfate, potassium metabisulphite)
 Use of antioxidants (e.g., tocopherol (Vitamin E), Ascorbic acid (Vitamin C), Citric
acid, Tartaric acid, EDTA, thioglycolic acid etc.).
 3-Complex chemical reactions:
▪ It includes dimerization, polymerizataion, racemization etc.

▪ In case of amoxicillin, besides hydrolysis, dimerization also takes place in concentrated

solutions.

▪ Polymerization is major mechanism of degradation in disinfectant glutaraldehyde at low pH.

▪ In racemization, an optically active substances loses its optical activity without change in its
chemical composition. e.g., levo adrenaline is 15-20 times more active than dextro
adrenaline. Racemization usually causes degradation with 1 st order kinetic principles.

▪ Suitable storage conditions prevent complex reactions.

 4-Photolysis:
 When molecules are exposed to electromagnetic radiation they absorb light (photons) at a
characteristic wavelength which causes increase in energy which can:
1. Be retained or transferred
2. Be converted to heat
3. Cause decomposition
4. Result in light emission at a new wavelength
 In natural sun light only higher energy UV rays range cause photo degradation of drugs.

Protection against Photolysis:

 Use of amber coloured bottles.
 Storage of drug products in dark places.
 Packaging in cardboard cartons to hinder light exposure.
 Coating of tablets with polymeric films.
 3-Microbiological degradation:

▪ Contamination from microorganisms is a big problem for all formulations

containing moisture but it can be a bother in solid dosage forms also if some
natural polymers are used because many natural polymers are fertile
sources of microorganisms.

▪ In a hygienic manufacturing environment where “Quality Assurance” is a

pre-requisite as per the GMP procedures, there are definite procedures to
prevent microbial contamination in all formulations.
Sources of Microbial Contamination:
1. Water used in manufacturing
2. Air in the working environment
3. Raw materials, containers and closures if they are not previously sterilized
4. Personnel may bring different microbes with them
5. Instruments and apparatus under poor sanitation conditions
Protection:
▪ Addition of antimicrobial preservatives
▪ Proper storage conditions
▪ Avoiding contamination especially for sterilized products
 References:
▪ Bentley’s textbook of Pharmaceutics An Adaptation by Sanjay K.Jain

▪ Aulton’s Pharmaceutics The Design and Manufacture of Medicine 3 rd ed.

▪ Theory and practice of Physical Pharmacy by Gaurav Kumar Jain and

Farhan Jalees Ahmad.