OBJECTIVES:

⦿1. Define related terms in stability testing and

enumerate the different types of drug stability
and how to maintain them
⦿2. Differentiate between long-term and
accelerated stability tests
⦿3. Solve problems involving chemical kinetics
to determine the shelf life and half life of a
medicinal agent
⦿ 4. Demonstrate appropriate skill in
handling and manipulating modern
instrument by performing experiment
⦿ 5. Predict with accuracy the expiration date
of a drug preparation using the knowledge
from this course
⦿ as well as diligence in performing assigned
task for the week
 OUTLINE:

Stability Testing
⦿Definition of Terms/Types of stability
⦿Factors affecting drug stability
⦿Long Term & Accelerated Stability Tests
⦿Chemical kinetics
⚫ zero order reaction
⚫ first order reaction
⚫ second order reaction
 STABILITY
⦿ Ability of a formulation in a specific
container to remain within its physical,
chemical, therapeutic and toxicological
specification
Shelf Life
The time from the manufacture of the
formulation until its chemical or biological
activity is not less than 90% of the
labeled potency
Why 90% ?
1. Loss of activity or potency of active
ingredient
2. Amount of degradation product
3. For cosmetics – retention of physical
qualities
Period of Stability
⦿ The time from the manufacture of the
formulation until its chemical or
biological activity is nlt 90% of the
labeled potency

⦿ Also called SHELF LIFE – remain

physically, chemically and biologically
stable
Important parameters for drug
products:
1. Loss of activity or potency of
active ingredient
2. Amount of degradation
product
3. For cosmetics – retention of
physical qualities
Types of Stability Testing
⦿ Accelerated Stability Testing
⦿ Real time or Long Term Stability
Testing
⦿ Alternate stability testing
Accelerated Stability
Testing
⦿ Use to predict product stability within
short period of time
US FDA shows acceptance of
Accelerated method is as follows:
1. Three months acceptable data at 37-40
C/ 75% RH can be extrapolated to a 2
–year expiry date
2. If a 2-year controlled room temp (RT)
samples are available, up to 2 more
years could be added to the expiry date
(four years total) by storing the 2 year
RT samples at 37 C/ 75% RH for 3
mos.
Factors affecting Stability
1- Environmental factors
- Temperature - Light
- Oxygen - Moisture
- Carbon dioxide

2- Drugs or excipients in the dosage form

Particle size of drug
pH of the vehicle

3- Microbial contamination
4- Trace metal Contamination
5- Leaching from containers
Shelf Life
⦿ Is calculated using Arrhenius equation or
by regression line analysis
Expiration date
⦿ Direct application and interpretation of
the knowledge gained from stability
testing
Expiration date appears on labeling as
month and year – refers to the last day
of the indicated month
 OVERAGES
Voluntary introduction of specific excess
during manufacture that are unstable by
nature and difficult to stabilize to maintain
during their period of use

⦿ Problem of declining potency

⦿ Use of overages to extend shelf life except
for expensive substance
⦿ Can also mean overdosage
Overages are justifiable when:
1. Labile active ingredient cannot possibly
be standardized
2. Would not present overdosage (narrow
therapeutic range)
3. Overage therapeutically
Proposed rules on overages
for vitamins are:
1. A loss of nmt 10% of the labeled
potency is considered normal at the
end of the term of validity of the
product
2. The added overage is limited to nmt
30%
⦿ A 15% decrease in potency of antibiotic
is admissible for unstable antibiotics
⦿ Overages:
dry dosage forms – 15%
fluids – 20%
ointments, supp., aerosols, creams –
25%
 Types:
⦿Manufacturing Overage – added to a
preparation to compensate loss during
manufacturing of the preparation

⦿Stability
Overage – is the excess added to
a preparation to extend its shelf life
Formulation:
⦿ Non elegant formulation – 1st stage
⦿ Formulation development – 2nd stage
- support Phase II and III
- prototype for the commercial
product
⦿ Final formulation – final stage
- for commercial pharmaceutical
product
Formulation Process
Types of stability:
1. Chemical
2. Physical
3. Microbiological
4. Therapeutic
5. Toxicological
Types of Stability Conditions Maintained
throughout Shelf Life

1. CHEMICAL - Chemical integrity and

labeled potency retained

2. PHYSICAL - Physical prop.

(appearance,palatability,
uniformity) retained
3. MICROBIOLOGICAL - Sterility or resistance to
microbial growth

4. THERAPEUTIC - Effects remain

unchanged

5. TOXICOLOGICAL - No significant increase

in toxicity occurs
Physical stability

Formulation Likely physical Effects

instability problems

Oral solutions 1- Loss of flavour Change in

2- Change in taste smell or
3- Presence of off flavours feel or
due to interaction with taste
plastic bottle
4- Loss of dye
5- Precipitation
6- discoloration
Physical stability (Cont.)
Formulation Likely physical Effects
instability problems
Parenteral 1. Discoloration due to photo Change in
chemical reaction or appearance
solutions oxidation and in
2. Presence of precipitate due to bio-availabil
interaction with container or ity
stopper
3. Presence of “whiskers”
4. Clouds due to:
(i) Chemical changes
(ii) The original preparation of a
supersaturated solution
Physical stability (Cont.)
Formulation Likely physical Effects
instability problems

Suspensions 1- settling 1-Loss of drug

2- caking content
3- crystal growth uniformity in
different doses
from the bottle

2- loss of
elegance.
Physical stability (Cont.)
Formulation Likely physical Effects
instability problems
Emulsions 1- Creaming 1- Loss of drug
2- coalescence content
uniformity in
different doses
from the bottle

2- loss of
elegance
Official Storage Conditions:
Cold: Not exceeding 8 oC
ref 2 to 8 oC
freezer -25 to -10 oC

Cool 8 to 15 oC

Room temp (RT) Working area

Controlled RT 20 to 25 oC

Warm 30 to 40 oC

Excessive heat Above 40 oC

Stability Protocols:
CONDITIONS TEMP/RELATIVE MINIMUM
HUMIDITY (RH) REQ. TIME
AT
SUBMISSION
Long term/ Real 25 oC ± 2 oC / 12 mo
time Testing 60% ± 5% RH

Accelerated 40 oC ± 2 oC / 6 mo
Testing 75% ± 5% RH

*Alternate 30 oC ± 2 oC / 12 mo
Testing 65% ± 5% RH
Alternate stab test
* Required if significant change occurs
during 6 mo. storage under conditions of
accelerated testing
Geographic regions of the world are
defined by CLIMATIC ZONES:
ZONE 1 TEMPERATE GREAT BRITAIN,
CANADA
ZONE II SUBTROPICAL US, JAPAN

ZONE III HOT AND DRY IRAN, IRAQ

ZONE IV HOT AND INDONESIA,

HUMID PHILIPPINES
Storage Condition Products Testing
Frequency

Real Time/ Long Term

⦿ 0, 3, 6, 9, 12, 18, 24 months and
annually through the proposed shelf-life

Accelerated
⦿ 0, 3 and 6 months
According to ICH (International Conference on
Harmonisation) for Pharmaceuticals for
Human Use: (Europe, Japan, US)
⦿ Form a guideline that provides indication on
the requirements for Stability Testing of New
Drug Substances and Products
⦿ Incompatibles are substances which
have opposite medicinal properties, or
substances when mixed together,
react chemically to produce other
substances.
⦿ An understanding of incompatibilities
can save the pharmacist valuable time
in compounding as well as ensure the
therapeutic efficiency of the products.
⦿ Incompatibilities range from minor to
dangerous.
⦿ Incompatibilities are divided
into three classes:
✔ Therapeutic
incompatibilities
✔ Physical incompatibilities
✔ Chemical incompatibilities
⦿ Physical – formation of precipitate, color
change
⦿ Chemical – visible change is not
necessarily observed; requires
trained, knowledgeable
personnel to recognize it
⦿ Therapeutic – undesirable pharmacologic
interaction between 2 or more
ingredients that leads to:
Consequences
1. Potentiation of the therapeutic
effects of ingredients
2. Destruction of effectiveness of one
or more of the ingredients
3. Occurrence of a toxic manifestation
within the patient
 Therapeutic Incompatibility
⦿They exist when the response to
one or more drug is of different
nature or intensity than that
intended by the prescriber.
⦿When circumstances produce a
feeling of doubt on the part of the
pharmacist, the prescribing
physician should be consulted.
 1. Synergistic (or additive) combination
⦿When certain drugs having the same
pharmacologic action are prescribed together, the
combined action they produce is greater than the
sum of their individual actions.
⦿Such combinations should be in reduced amount.
⦿The synergistic action will sometimes be desired
to decrease toxicity or reduce cost of prescription
⦿ Synergistic (cont.); examples of useful
synergistic effect
⦿ Combinations of sulfonamides show less
nephrotoxicity than a single sulfonamide.
⦿ Neomycin – erythromycin combination has
a broader spectrum than either antibiotic
alone.
⦿ Aspirin – codeine combination reduces the
amount of expensive codeine required.
2. Antagonistic combinations
When two or more drugs having opposite
pharmacological action are prescribed
together, their actions cancel each other and
the resulting prescription will have no
therapeutic action.
Stimulants with sedatives (hypnotics) e.g.
caffeine with chloral hydrate.
Purgatives with antidiarrheals e.g. MgSo4 with
tannins
Acidifiers with alkalinizers e.g. aspirin with
AL(OH)3.
 3. contraindication
⦿Are drugs when prescribed together
may lead to increased toxicity or
decrease activity.
⦿Tetracycline is contraindicated with
any drug containing Ca2+ ions as it
form non-absorbable complex.
⦿Inactivation of sulfa drugs by procaine
HCI.
 Physical Incompatibility:
⦿ Physical incompatibilities are often called
pharmaceutical incompatibilities and are evidence
d by the failure of the drugs to combine properly.
⦿ These incompatibilities produce a mixture which is
unacceptable in appearance and taste and may
result in non-uniform dosage form.
Types of physical
incompatibility
Incomplete solution
Gums are insoluble in
alcohol.
resins are insoluble in water
Precipitation
Resins are precipitated from
alcoholic solution when water is
added
Camphor and volatile oils are
salted out from their aromatic
water when soluble salts are
added
Other physical
incompatibilities
⦿ Separation of immiscible
liquids
⦿ Eutexia
⦿ Incorrect form
prescribed/dispensed
⦿ Adsorption
Physical Stability:
1. Appearance – elegant
2. Uniformity – products in multiple dose
containers; proper amount in each
dose
3. Availability – breakdown in the physical
system can lead to non-availability of
medicament
Chemical Incompatibility:
⦿ This type of incompatibility exists when
agents are prescribed that react chemically
when mixed, altering the composition of
one or more of the constituents.
⦿ Examples;
- formation of precipitate
- evolution of gas
- colour change
Chemical stability
implies:
⦿ The lack of any decomposition in
the chemical moiety that is
incorporated in the formulation as
the drug, preservatives or any other
excipients.
⦿ This decomposition may influence
the physical and chemical stability
of the drug
Deterioration caused by:

oxidation
reduction
hydrolysis
racemization
epimerization
Chemical Reactions
 1. Oxidation – reduction
⦿ Oxidation is the prime cause of product
instability
⦿ N2 and CO2 displaced headspace air in
pharmaceutical container to minimize
deterioration by oxidation
⦿ Heavy metals (cupric, ferric) catalyze oxidation
reactions
⦿ Inhibited by antioxidants (Na metabisulfite, Na
thiosulfate, ascorbic acid, etc.)
 The ideal antioxidant should be:
⦿Stable
⦿Effective over a wide pH range
⦿Soluble
⦿Colorless
⦿Nontoxic
⦿Nonvolatile
⦿Nonirritating
⦿Effective in low concentration
⦿Thermostable
⦿Compatible with other ingredients
2. Hydrolysis

⦿ Drugs containing esters (cocaine,

physostigmine, aspirin, tetracaine,
procaine, methyldopa), amide
(dibucaine), lactam (penicillin,
cephalosporin)
⦿ Ex. acetyl ester in aspirin is
hydrolyzed to acetic acid and salicylic
acid in the presence of moisture
⦿ Ex. Acetaminophen (paracetamol) to
p-aminophenol and acetic acid

⦿ Ampicillin to aminobenzyl penillic acid

⦿ Amoxicillin to penicilloic acid

3. Decarboxylation
⦿ Carboxylic acids such as
p-aminosalicylic acid lose CO2 from
carboxyl group when heated producing
reduced pharmacological potency
⦿ Store in Cool Place
⚫ Main cause is heat
⦿ Amber colored bottle
4. Racemization
⦿ Process of changing from an optically
active compound into optically inactive
mixture
5. Epimerization
⦿ Tetracycline undergoes (exposed to pH
higher than 3)
⦿ The epimer of tetracycline has little or no
antibacterial activity
⦿ The epimer differ in configuration around
single carbon atom
 CORRECTIVE MEASURES
⦿Addition of an ingredient that does not alter the
therapeutic value.
⦿Change of an ingredient. Minor changes such as
a soluble form of an ingredient for an insoluble
form are included.
⦿Change of a solvent.
⦿ The utilization of special techniques in
compounding, such as treating each
ingredient separately.
⦿ Omission of an agent that has no
therapeutic value, or that may be
dispensed separately.
Photochemical Reactions:
⦿ Photolytic degradation
⦿ Affected chemically by radiation (UV)
⦿ Ex. Nifedipine, nitroprusside,
riboflavin
⦿ Use colored glass container (yellow
green glass and amber against UV)
Ionizing Radiation:

⦿ Particularly gamma rays is used for

sterilization of certain pharmaceutical
products but still causes chemical
degradation
Stabilization of drugs against hydrolysis,
oxidation and photolysis

1- Temperature
All the drug products are stored at suitable
temperatures to avoid thermal acceleration of
decomposition. Three varieties of temperatures are
suggested for storage of drug products. Room
temperature, cool storage and cold storage.

2- Light
Light sensitive materials are stored in ambered
colour bottles.
Stabilization of drugs against
hydrolysis, oxidation and photolysis
3- Humidity
Packing materials are chosen (usually glass
and plastic) to prevent exposure of drug
products to high humid condition.

4- Oxygen
Proper packing keeping the oxygen content
of the solution less and leaving very little
head space in the bottle above the drug
products are methods to fight against
oxidation.
Stabilization of drugs against hydrolysis,
oxidation and photolysis

Antioxidants commonly used for

Aqueous systems Oil systems
Sodium metabisulfite Ascorbyl palmitate
Sodium thiosulfate Butylaled hydroxy toluene
Ascorbic acid Butylated hydroxy anisole
Stabilization of drugs against
hydrolysis, oxidation and photolysis

5- Chelating Agents
Chelating agents form complexes with heavy metal
ions and prevent them from catalyzing oxidative
decomposition.
e.g. ethylenediamine tetracetic acid (EDTA) derivatives
and salts, citric acid and tartaric acid.

6- Solvents
By the addition of a suitable solvent hydrolysis rate
may be decreased.
Microbiological stability

⦿ Microbiological stability implies that:

The formulation has not suffered from any
microbiological attack and is meeting the standards
with respect to lack of contamination/sterility.
Microbiological stability

Sources of Microbial Contamination:

Water gram-negative groups: Pseudomonas,

Xanthamonas, Flavobacterium

Air Mould spores: Penicillium, Aspergillus

Bacterial spores: Bacillus spp. Yeasts

Raw materials Micrococci

Starches Coliforms
Pigments Salmonella
Sources of Microbial Contamination

Gums Actinomyces

Animal products Salmonella, Coliforms

Personnel Coliforms, Staphylococci, Sterptococci

To prevent contamination to the
formulation during storage

(1) suitably designing the containers

(2) usually using single dose containers

(3) sticking to proper storage conditions

(4) adding an antimicrobial substance as

preservative.
Preservatives used in pharmaceutical
preparations:
Preparation Preservative Concentration
% w.v
Injections Phenol 0.5
Cresol 0.3
Chlorocresol 0.1
Eye drops Chlorhexidine 0.01
acetate 0.01
Benzalkonium
chloride
Mixtures Benzoic acid 0.1
Methyl paraben 0.1
Alcohol 12-20
Preservatives used in pharmaceutical
preparations:

Preparation Preservative Concentration

% w.v

Creams Parabens 0.1-0.2

Chlorocresol 0.1
Tablets Methylparaben 0.1
Pharmaceutical
Containers
⦿ WELL CLOSED – protect contents from
extraneous solids
⦿ TIGHT CONTAINER – protect contents
from liquid, solids or vapors
⦿ HERMETIC CONTAINER – impervious
to air or any other gases
Glass:
⦿ Most widely used
⦿ Disadvantage:
leach of alkali and insoluble flakes but
can be offset by the choice of
appropriate glass
Glass Types:
TYPES DESCRPTION USES TYPE OF TEST

I Borosilicate glass parenterals Powdered

glass
II Treated soda Used when Water attack
lime glass stability data (titrated w/
demonstrates H2SO4)
its suitability
III Soda lime glass Oral or Powdered
topical use glass
NP Oral or Powdered
topical use glass
Include table for purpose and
limits of 0.02 N sulfuric refer to
USP 29
Glass Tests
Determine resistance to water
attack of new (not previously
used)
Degree of attack is determined
by the amount of alkali
released from the glass
Glass tests:
Water Attack Test:
1.Fill bottle with high purity water
2.Autoclave
3.Place the liquid specimen into flask
4.Add methyl red
5.Titrate with 0.02 N H2SO4
Powdered Glass Test: Sample
Preparation
Powdered Glass Test: Sample
Preparation
Powdered Glass Test: Sample
Preparation
Powdered Glass Test: Auxiliary
Preparation
⦿ Prepare Auxiliary items
⚫ conical flask and glass cap
⦿ Fill flask with High Purity Water
⦿ Submerge cap in High Purity
Water
⦿ Heat in bath at 90oC for 24 hours
Powdered Glass Test
Powdered Glass Test
Plastics:
⦿ Polyethylene
⦿ Polystyrene
⦿ Polyvinylchloride (PVC) –
⦿ Polypropylene
Major disadvantage of plastic:
⦿ two way permeation or breathing
through container wall;
⦿ volatile/ perfume/ flavoring are
permeable through plastics;
⦿ Oxygen and carbon dioxide in air
migrate
⦿ Metals

Various alloys and aluminium tubes may be

utilized as containers for emulsions,
ointments, creams and pastes.

Limitation: They may cause corrosion and

precipitation in the drug product.

Overcome: Coating the tubes with

polymers may reduce these tendencies.
⦿ Rubber

Rubber also has the problems of

extraction of drug ingredients and
leaching of container ingredients.
REACTION KINETICS
 Applications:
⦿Chemical reactions such as
decomposition of medicinal
compounds
⦿Processes of drug absorption,
distribution and elimination from the
body
⦿Shelf life determination.
Shelf life determination:
In determining the shelf life of a preparation, tests
are carried out on the active ingredient, the
additives and the finished product to determine:
⦿Whether decomposition will occur
⦿The type of decomposition
⦿Factors that affect the rate of decomposition such
as light, air, moisture, temperature, etc.
⦿The influence of formulation additives
⦿The rate at which breakdown occurs.
Order of Reaction
⦿ Manner in which the rate of reaction varies
with the concentration of the reactants
⦿ Some drug degradation processes can be
treated as either First or zero order
processes
 Zero order kinetics
⦿Zero order kinetics is where the
plasma concentration of a drug
decreases at a constant rate. A graph
of this will show a linear relationship
between time from peak concentration
and plasma concentration. The
situation may be contrasted with First
order kinetics and Second order
kinetics.
 Examples of drugs undergoing zero
order metabolism
⦿Phenytoin, Phenylbutazone
⦿Warfarin
⦿Heparin
⦿Ethanol
⦿Aspirin
⦿Theophylline, Tolbutamide
⦿Salicylates
Graph for Zero order:
Graph for First Order:
ORDER EQUATION HALF LIFE SHELF LIFE
(t ½) (t90)

ZERO C = -k0 t + C0 0.5 C0 0.1 C0

ORDER
k0 k0

FIRST K = 2.303 log C0 0.693 0.105

ORDER
t C k1 k1
Where:
C0 = original conc.
C = conc. remaining
k = rate constant
st
k1 = rate constant at 1 order
k0 = rate constant at zero order

Units: k = conc./ time

Problem Solving
Strategies
Strategy for solving Kinetics
problems:
⦿ 1. Determine k from the point-slope
formula
⦿ 2. Determine half life
⦿ 3. Determine shelf life
⦿ 4. Which equation?
Strategy for determining the
reaction order:
⦿ 1. Plot Log(A) vs. Time
⚫ A linear plot will indicate a 1st order
reaction
⦿ 2. Plot A vs. Time
⚫ A linear plot indicates zero order
How are half life, reaction rate,
and initial concentration
related for zero order
reactions?
How are half life, reaction rate,
and initial concentration
related for first order
reactions?
Solid state versus solution
stability:
⦿ Generally, chemical reactions proceed
more readily in liquid state than in solid
state
⦿ Serious stability problems are more
commonly encountered in liquid
medicines e.g. order of dosage form
stability is generally: solution <
suspension < tablet.
Problems:
1. A drug suspension (125 mg/mL) decays
by zero order kinetics with a reaction
rate constant of 0.5 mg/mL/hr. What is
the concentration of intact drug
remaining after 3 days (72 hrs)?
2. How will it take for the suspension to
reach 90% of its original concentration?
 3. A solution of a drug was freshly prepared at
a concentration of 300 mg/mL. after 30
days @ 25 C, the drug concentration in
the solution was 75 mg/mL.
a. Assuming 1st order kinetics, when will the
drug decline to one half of the original
conc.?
b. Assuming zero order kinetics…?
4. Determine : % potency Time (mo)
a. whether the 100 0
decomposition is 94.9 1
zero or first order
89 2
b. The decomposition
84 3
rate constant and
half life 70.6 6
c. Shelf life (t90) 49.9 12
35.2 18
5. A pharmacist dissolved an Time (hr) Conc
antibiotic into 100 ml water
and placed soln in a ref. At
(mg/ml)
various time interval, the 0.5 84.5
pharmacist removed a 10 mL
aliquot and measured the 1.0 81.2
amount. The ff. data are
obtained: 2.0 74.5
4.0 61.1
6.0 47.5
8.0 34
Temperature Effects
 ACTIVATION ENERGY (Ea):
Interpretation:
1. As the reaction proceeds from reactants to
products, the system must pass through a
state whose energy is greater than that of
initial reactants
2. This “barrier” is what prevents the
reactants from becoming products
3. The activation energy is a measure of this
barrier
Ea for some Drug Decomposition rxn:
COMPOUND REACTION Ea (kcal/mol)
Ascorbic acid oxidation 23
aspirin hydrolysis 14
atropine hydrolysis 14
benzocaine hydrolysis 19
Chloram hydrolysis 20

Thiamine hydrolysis 20
epinephrine oxidation 23
Temperature – Rate of most chemical
reactions increase with rise in temperature
up to 2 to 3 times with each 10° rise in
temperature.

The relationship is expressed by Arrhenius

equation:
Generally…
⇧Temp - ⇧ rate of reaction

Log (k1/k2) = [-Ea/(2.303R)] [(1/t1)-(1/t2)]

R = gas constant = 1.987 cal/Kmol

Formula:

Log k2 = -Ea (1- 1)

k1 2.303 (R) T 2 T1

Where:
R – Gas constant = 1.987 cal K-1 mol-1 or
1.987 cal/Kmol
1. Consider the sulfacetamide monograph,
the 1st order rate constant is 9 x 10-6 s-1
at 120 C. The activation energy is 22.9
kcal/mole. Calculate the shelf life @25
C?
Shelf-life Estimation
Methods:
⦿ Reasonable estimates of shelf life to be
made when the product is stored under
non standard condition of temp.
⦿ Effects of temp on shelf life
⦿ Assuming Ea values are not available,
use Q10 values of 2, 3 and 4 to estimate
shelf life
Q10
⦿ Method allows the pharmacist quickly to
calculate the estimates of shelf life for a
product stored in different conditions
⦿ where t90T2 is the estimated shelf life
⦿ t90T1 is the shelf life at a given temperature
⦿ ∆T is the difference in temperature between
T1 and T2 (i.e. T2 – T1)
⦿ Increase in ∆T will decrease shelf life while
a decrease in ∆T will increase shelf life
⦿ Formula:
Q10 FORMULA
EXAMPLE
⦿ An antibiotic solution has a shelf life of
48hrs at refrigerator. Using a Q value of
3, what is the estimated shelf life in
room temp?
1. The expiration period for a reconstituted
product is 18 h at room temp. estimate the
expiration period when the product is
stored in the ref .
2. A newly reconstituted product is labeled
to be stable for 24 hrs in a ref. what is
the estimated shelf life at room
temperature?
3. A product has a t90 of 48 hrs. in a ref. (5
C). What would t90 be at room temp.(25
C)?