Prof.Dr.Mowafaq M.

Ghareeb
Pharmaceutical Kinetics and Stability

Pharmaceutical Kinetics and Stability

Objectives
At the conclusion of this course the student should be able to:

1. Know the general types of degradation

2. Define reaction rate, reaction order, and molecularity.
3. Understand Mechanism of degradation; simple and complex reactions
4. Understand and apply apparent zero-order kinetics to the practice of pharmacy.
5. Calculate half-life and shelf life of pharmaceutical products and drugs.
6. Understand Michaelis–Menten (nonlinear) kinetic behavior and linearization
techniques.
7. Understand the basis for transition-state theory and its application to chemical
kinetics.
8. Describe the influence of temperature, ionic strength, solvent, pH, and dielectric
constant on reaction rates.
9. Calculate the increase in rate constant as a function of temperature (Q10).
10. Interpret pH–rate profiles and kinetic data.

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Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability
Importance of kinetic study

The purpose of stability testing is to provide evidence on how the quality of a

drug substance or drug product varies with time under the influence of a variety of
environmental factors, such as temperature, humidity, and light, and to establish a retest
period for the drug substance or a shelf life for the drug product and recommended
storage conditions. Although the pharmaceutical scientist plays a critical role in
determining the stability of pharmaceuticals, practicing pharmacists should be able to
interpret this information for their patients. This course introduces the rates and
mechanisms of reactions with particular emphasis on decomposition and stabilization of
drug products. It is essential for pharmacists and pharmaceutical scientists to study,
understand, and interpret conditions of instability of pharmaceutical products as well as
to be able to offer solutions for the stabilization of these products.

Pharmaceutical manufacturers routinely utilize the principles covered in this

course; however, with the resurgence of pharmaceutical compounding, it is essential for
practicing pharmacists to understand drug product stability as well. If a community
pharmacist is asked to compound a prescription product, there are many factors that he or
she must consider. The pharmacist must recognize that alterations in stability may occur
when a drug is combined with other ingredients. For example, if thiamine hydrochloride,
which is most stable at a pH of 2 to 3 and is unstable above pH 6, is combined with a
buffered vehicle of, say, pH 8 or 9, the vitamin is rapidly inactivated. Knowing the rate at
which a drug deteriorates at various hydrogen ion concentrations allows one to choose a
vehicle that will retard or prevent the degradation. Patients expect that products will have
a reasonable shelf life.
Even though pharmaceutical manufacturers label prescription and over-the-
counter drug products with expiration dating to guide the patient/consumer in these
matters, patients may store these products in a bathroom medicine cabinet where the
humidity and temperature are higher than the typical storage place for medications.
How does this affect the shelf life of the product? A community pharmacy
practitioner should be able to understand this and advise patients on these matters.
The experimental investigation of the possible breakdown of new drugs is not a simple
matter. Applications of chemical kinetics in pharmacy result in the production of more
stable drug preparations, the dosage and rationale of which may be established on sound
scientific principles. Thus, as a result of current research involving the kinetics of drug
systems, the pharmacist is able to assist the physician and patient regarding the proper
storage and use of medicinal agents. This course brings out a number of factors that bear
on the formulation, stabilization, and administration of drugs. Concentration,
temperature, light, pH, and catalysts are important in relation to the speed and the
mechanism of reactions and will be discussed in turn.

Example of kinetic application in pharmacognocy, an important part of quality

control of herbal drug preparations is the evaluation of the chemical stability of a finished
product during the storage period. Measuring chemical stability is very challenging due
to the complexity of a plant extract, which may contain thousands of different
compounds.

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Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability
Moreover, the presence of enzymes like glycosidases, esterases or oxidases plays
an important role in the breakdown of secondary plant metabolites. This is of particular
significance for liquid herbal products.
Example of kinetic application in clinical pharmacy: Absorption, distribution and
elimination processes that are associated with the role of absorption of a drug into the
body, the rate at which the drug is subsequently distributed through the body and the rate
at which it is removed from distribution by factors such as metabolism, storage within a
body organ or fat, and by excretory routes.
Using chemical reaction kinetics to predict optimal antibiotic treatment strategies by pia
abel zur wiesch et al is another example of application.
Example of kinetic application in pharmacology; Nitric oxide, NO, is central to
many physiological processes including regulation of blood pressure and nerve signal
transmission. Enzymes in endothelial cells and in the brain of mammals continuously
synthesize it—generally in low and carefully regulated concentrations. The well–known
reaction of NO with oxygen to produce toxic nitrogen dioxide, NO2, has a rate which is
bimolecular in NO. High concentrations of NO, as are found often in industrial plants or
cigarettes, react rapidly with oxygen to produce toxic NO2. However, the half-life of NO
at low NO concentrations as found in solutions and gases occurring in blood vessels,
brains, and lungs is sufficiently long for biochemical purposes. Kinetics, then, determines
the harmful versus helpful aspects of nitric oxide. At concentrations below 80 ppm NO is
used in hospitals for lung vasodilation of preterm newborns and patients with pulmonary
distress.

Example of kinetic application in biochemistry; Enzymes are effectors of chemical

change. Through their action, enzymes bring about the transformation of one chemical
substance into another. Like all change, the chemical change brought about by enzymes
involves a temporal aspect that can be expressed as the rate at which the change occurs.
The systematic study of these rates defines the biochemical field known as enzyme
kinetics.

Life depends on dynamic biochemical processes, and understanding these reactions

requires knowledge of physical constants called rate constants. In addition to kinetic
study of enzym work, there are many cellular reactions do not involve catalysis or
chemical cleavage of a substrate. Examples include proteins binding to other proteins,
nucleic acids, membranes, or small molecules.

The first quantitative study in chemical kinetics has been done by German scientist
Ludwig Ferdinand Wilhelmy (1812–1864) in 1850 that used polarimetry to investigate
the acid-catalyzed conversion of sucrose. In this early study, Wilhelmy recognized that
the reaction rate was proportional to the concentration of sucrose and acid.

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 Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability

General types of degradation

There are three main categories of drug degradation include chemical, physical, and
microbial types as shown in this table;

Category Description Examples

I-Chemical Breakage or formation of covalent bonds resulting in, -Ester hydrolysis
unstability for example, loss of potency. -Amide hydrolysis
-Lactam hydrolysis
-Oxidation
II-Physical No breakage or formation of covalent bonds. Such -Crystallization of
unstability changes can lead to changes in drug solubility or amorphous drugs
changes in the appearance of the drug product. -Changes in crystal
forms
-Loss of crystal water
III-Microbial Microbial contamination -Aqueous eye drop
unstability solutions
-Parenteral solutions

I-Pathways of Chemical Degradation:

1-Hydrolysis
2- Oxidation
3-Dehydration.
4-Isomerization and racemization
5- Decarboxylation and elimination
6- Photodegradation
7- Polymerization
8- Drug -excipient interaction .

1-Hydrolysis:
hydrolysis is one of the most common reactions seen with pharmaceuticals. Hydrolysis
indicates cleavage of chemical bonds by the addition of water. How can we tell whether a drug is
likely to be susceptible to this type of degradation? If the drug is a derivative of carboxylic acid or
contains functional groups based on this moiety, for example an ester, amide, lactone, lactam, imide or
carbamate, then we are dealing with a drug which is liable to undergo hydrolytic degradation.The
hydrolytic reactions involve nucleophilic attack of the labile groups and the most common substrates
are those that contain the acyl group as shown in the following table:

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Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability

Drugs that contain ester linkages include acetylsalicylic acid (aspirin),

physostigmine, methyldopate, tetracaine and procaine. Ester hydrolysis is usually a
bimolecular reaction involving acyl–oxygen cleavage. For example, the hydrolysis of
procaine is shown in Scheme below

The hydrolysis of amides involves the cleavage of the amide linkage as for example, in
the breakdown of the local anaesthetic cinchocaine (Scheme below).

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Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability
2- Oxidation

After hydrolysis, oxidation is the next most common pathway for drug breakdown.
Oxygen is one of the abundant elements in the environment. Upon exposure to oxygen,
pharmaceuticals that are not in their most oxidized state may decompose.
Oxidation/reduction reactions involve the transfer of electrons or the transfer of oxygen
or hydrogen from a substance. There should be the drug molecules to be oxidized (i.e.,
reducing agents) and other substances to be reduced (i.e., oxidizing agents) in a same
system. Oxidation of inorganic and organic compounds is easily explained by a loss of
electrons and the loss of a molecule of hydrogen, respectively,as:

When an oxidation reaction involves molecular oxygen, the reaction occurs

spontaneously under mild conditions. It is known as
autooxidation. In an autooxidation process, free radicals, formed by thermal or photolytic
cleavage of chemical bonds (e.g., peroxide, ROOH) or redox processes with metal ions
present in raw material impurities, are involved

The free radical formed, RO* , reacts with oxygen to produce a peroxide radical, and the
reaction propagates as:

The free radical reaction continues until all the free radicals are consumed or
destroyed.
Oxidation mechanisms for drug substances depend on the chemical structure of the drug
and the presence of reactive oxygen species or other oxidants. Catechols such as
methyldopa and epinephrine are readily oxidized to quinones, as shown in Scheme below

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Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability

3-Chemical Dehydration
Sugars such as glucose (Scheme) and lactose are known to undergo
dehydration to form 5-(hydroxymethyl)furural. Erythromycin is susceptible to acid
catalyzed dehydration as shown in Scheme

4- Isomerization and racemization

Isomerisation is the process of conversion of a drug into its optical or geometric

isomers. Since the various isomers of a drug are frequently of different activity, such a
conversion may be regarded as a form of degradation, often resulting in a serious loss of
therapeutic activity. For example, the appreciable loss of activity of solutions of
adrenaline at low pH has been attributed to racemisation – the conversion of the
therapeutically active form, in this case the levorotary form, into its less active isomer.

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Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability
In acidic conditions the tetracyclines undergo epimerisation at carbon atom 4 to
form an equilibrium mixture of tetracycline and the epimer, 4-epi-tetracycline
(Scheme )The 4-epi-tetracycline is toxic and its content in medicines is restricted to not
more than 3%.

In aqueous solutions, L-(-)-Epinephrine undergoes specific acid catalyzed racemization

However, the main degradation route of epinephrine under aerobic conditions is

oxidation (the solution goes from colorless to pink) and other oxidation products. The
rate of oxidation increases with increasing pH, while the rate of racemization decreases
with increasing pH.

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Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability
The optimum stability of the aqueous epinephrine solutions has been estimated to be
between pH 3.0 and 3.8, at which the total rate of degradation (i.e., the rate of
racemization and the rate of oxidation) is at its minimum

5- Decarboxylation and Elimination

Drug substances having a carboxylic acid group are sometimes susceptible to
decarboxylation,as shown in Scheme. 4-Aminosalicylic acid is a good example. etodolac
is susceptible to decarboxylation.

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Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability
Other elimination reactions have been reported for various drug substances, as shown in
Scheme. Trimelamol eliminates its hydroxymethyl groups and forms formaldehyde.
Levothyroxine eliminates iodine.

6- Photodegradation

Light energy, similar to heat, provides the activation necessary for oxidation to
take place. After a drug substance has absorbed radiant light energy, it becomes an
unstable, excited species. The activated species either emits radiant light of a different
wavelength (no decomposition occurs) or decomposes. Oxidation very often accompanies
photodegradation in the presence of oxygen and light. The photolysis of a drug substance
may cause discoloration of the product and packaging materials in addition to chemical
degradation. The pathways of photolysis are generally very complex and depend on drug
molecules.
Many pharmaceutical compounds, including the phenothiazine tranquillizers,
hydrocortisone, prednisolone, riboflavin, ascorbic acid and folic acid, degrade when
exposed to light. As a result there will be a loss of potency of the drug, often
accompanied by changes in the appearance of the product, such as discoloration or
formation of a precipitate.

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Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability

7-Polymerisation
Polymerisation is the process by which two or more identical drug molecules
combine together to form a complex molecule. It has been demonstrated that a
polymerization process occurs during the storage of concentrated aqueous solutions of
aminopenicillins, such as ampicillin sodium. The reactive β- lactam bond of the
ampicillin molecule is opened by reaction with the side-chain of a second ampicillin
molecule and a dimer is formed (Scheme). The process can continue to form higher
polymers.
Such polymeric substances have been shown to be highly antigenic in animals and
they are considered to play a part in eliciting pencilloyl specific allergic reactions to
ampicillin in humans.

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Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability
8- Drug -excipient interaction

Reactions of Bisulfite, an Antioxidant, in the 1950s, it was reported that

epinephrine, a catecholamine, undergoes displacement of its hydroxy group by bisulfite,
as shown in Scheme.

II-Physical Degradation

Most studies on drug stability have focused on the chemical stability of drug
substances. However, the physical stability of drugs must also be considered. The
physical state of a drug determines its physical properties such as its solubility. Because
these properties in turn affect the efficacy and, potentially, the safety of
a drug substance, in addition, changes in the physical states of excipients or enabling
agents in a dosage form may affect the stability of pharmaceuticals.
Solids are broadly classified into two types; crystalline solids and amorphous
solids. A crystalline solid is a substance whose constituent particles possess regular
orderly arrangement e.g. Sodium chloride, sucrose, diamond etc. An amorphous solid is a
substance whose constituent particles do not possess a regular orderly arrangement e.g.
glass, plastics, rubber, starch, and proteins.
Components of pharmaceuticals (drug substances and excipients) exist in various
microscopic physical states with differing degrees of order. Examples are amorphous and
various crystalline, hydrated, and solvated states. With time, the drug or the excipient
may change from one state, usually unstable or metastable, to a more thermodynamically
stable state.
The following sections address the physical changes that can occur in drug substances
and excipients and describe factors affecting these physical changes

1. Crystallization of Amorphous Drugs

Attempts are often made to formulate poorly water-soluble drugs in their
amorphous state. This is because the solubility of amorphous materials is generally
higher than that of the same substances in their crystalline state. However, because of the
lower free energy of the crystalline state, amorphous substances tend to change to their
more thermodynamically stable crystalline state with time. Therefore, crystallization of
amorphous drug substances may occur during long-term storage and may lead to drastic
changes in the release characteristics of the drug and, hence, changes in its clinical and
toxicological behavior.

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Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability
Amorphous nifedipine, undergoes partial crystallization during storage under high-
humidity conditions. This change from a largely amorphous state to a partially crystalline
state resulted in altered dissolution and solubility behavior.

2. Transitions in Crystalline States

Polymorphs are different crystalline forms of the same drug. Because these forms
have different free energy or chemical potentials, depending on temperature conditions,
transitions between polymorphs occur. Polymorphic transitions during storage may alter
critical properties of drugs because the solubility and dissolution rate of drug substances
generally Polymorphs are different crystalline forms of the same drug.
Because these forms have different free energy or chemical potentials, depending
on temperature conditions, transitions between polymorphs occur. Polymorphic
transitions during storage may alter critical properties of drugs because the solubility and
dissolution rate of drug substances generally

3. Formation and Growth of Crystals

Molecules in a crystal, and the crystals themselves, should not be considered static.
Crystals can grow or decrease in size provided that there is a medium across which the
molecules can travel. This could be a liquid phase or a gaseous phase into which the
molecules can sublime. For example, drug substances and excipients in solid dosage
forms, such as tablets and granules, may recrystallize or sublime onto the surface of the
dosage form during storage. So-called, whisker, crystallization was observed in tablets of
ethenzamide and caffeine anhydride.

4. Vapor-Phase Transfers Including Sublimation

Pharmaceuticals containing components that sublime easily may undergo changes
in drug content owing to the sublimation of the drug substances or excipients. In the case
of nitroglycerin, which is a liquid with a significant vapor pressure, sublingual tablets
exhibited significant variations in drug content during storage owing to intertablet
migration through the vapor phase

5. Moisture Adsorption
Moisture adsorption is generally observed with solid pharmaceuticals. In addition to
effect of moisture adsorption on the chemical stability of pharmaceuticals, the moisture
adsorption during storage can also affect the physical stability of pharmaceuticals,
leading to changes in such properties as appearance and dissolution rate. Adsorption of
moisture is governed by the physical properties of the drug substance and excipients.

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Prof.Dr.Mowafaq M.Ghareeb
Pharmaceutical Kinetics and Stability
III- Microbial degradation
Microbiological stability Microbiological stability implies that: The formulation
should not suffer from any microbiological attack& is meeting the standards with respect
to lack of contamination/sterility. Microbial contaminants usually need to attack
formulation ingredients and create substrates necessary for biosynthesis and energy
production before they can replicate to levels where obvious spoilage becomes apparent.
Thus, for example, 106 microbes will have an overall degradative effect around 10 6 times
faster than one cell. However, growth and attack may well be localized in surface
moisture films or very unevenly distributed within the bulk of viscous formulations such
as creams.
Early indications of spoilage are often organoleptic, with the release of unpleasant
smelling and tasting metabolites such as ‘sour’ fatty acids, ‘fishy’ amines, ‘bad eggs’,
bitter, ‘earthy ’ or sickly tastes and smells. Products may become unappealingly
discoloured by microbial pigments of various shades. Thickening and suspending agents
such as tragacanth, acacia or carboxymethylcellulose can be depolymerized, resulting in
loss of viscosity and sedimentation of suspended ingredients. Alternatively, microbial
polymerization of sugars and surfactant molecules can produce slimy, viscous masses in
syrups, shampoos and creams, and fungal growth in creams has produced ‘gritty’
textures. Changes in product pH can occur depending on whether acidic or basic
metabolites are released, and become so modified as to permit secondary attack by
microbes previously inhibited by the initial product pH. Gaseous metabolites may be seen
as trapped bubbles within viscous formulations.

When a complex formulation such as an oil-in-water emulsion is attacked, a gross

and progressive spoilage sequence may be observed. Metabolism of surfactants will
reduce stability and accelerate ‘creaming’ of the oil globules. Lipolytic release of fatty
acids from oils will lower pH and encourage coalescence of oil globules and ‘cracking’ of
the emulsion. Fatty acids and their ketonic oxidation products will provide a sour taste
and unpleasant smell, while bubbles of gaseous metabolites may be visible, trapped in the
product, and pigments may discolour it.

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