STABILITY STUDIES

……… A measure of how a pharmaceutical product

maintains its quality attributes over time
STABILITY –
The capacity of a drug or product to remain within established
specifications of identity , quality, purity in a specific period of time.
OR
The capacity or the capability of a particular formulation in a specific
container to remain with in particular chemical , microbiological ,
therapeutically , and toxicological specifications.
OR
USP defines stability of pharmaceutical product as , “extent to which a
product retains with in specified limits and throughout its period of
storage and use ( i.e. shelf life).
Stability testing is used to:
 Provide evidence as to how the quality of the drug product varies with
time.
 Establish shelf life for the drug product.
 Determine recommended storage conditions.
 Determine container closure system suitability.
Why Stability studies are necessary ?
 Chemical degradation of the product leads to lowering of the concentration
of the drug in the dosage form.
Toxic products may be formed , due to chemical degradation of the active
ingredient.
Advantages of Stability studies
 Assurance to the patient Economic considerations Legal
requirement
 OBJECTIVES

1. To determine maximum expiration date/ shelf life.

2. To provide better storage condition.

3. To determine the packaging components.

4. To gather information during preformulation stage to

produce a stable product.
 Study Storage condition Minimum time
period covered
by data at
submission

Long Term 25º C ± 2º C 12 months

(Ambient) 60%RH ± 5%

Intermediate 30º C ± 2º C 6 months

(controlled) 60%RH ± 5%

Accelerated 40º C ± 2º C 6 months

75%RH ± 5%
 Kinetics
Motion or
Kinetics
movemen
t
Velocity, rate or
rate of change

Kinetics deals with the study of the rate at which

processes occur and mechanism of chemical reactions
It involves the study of rate of change and the way in which this rate is
influenced by the concentration of reactants, products, and other
chemical species that may be present, and by factors such as solvents,
pressure, and temperature.

Kinetics applies to:

Stability

Incompatibility,

Dissolution,

Absorption,

Distribution

Drug action at molecular level

Elimination processes
 WHY DO WE STUDY ABOUT KINETICS?

It gives an in light into the mechanism of changes involved

Allows a prediction of the degree of change that will occur

after a given time has elapsed.
 DRUG STABILITY
• The resistance of the drug to the various chemical, physical, and
microbiological reactions that may change the original properties of
the preparations during transport, storage and use.

• Quantitatively it is expressed as shelf life.

Shelf life
 is the time during which the medicinal product is predicted to
remain fit for its intended use under specified conditions of storage.

 It is the time from manufacture or preparation until the original

potency or content of the active ingredient has been reduced by 10%
[t10 or t90] which is the limit of chemical degradation
WHY DO WE STUDY ABOUT DRUG STABILITY?

Safety of the patient [toxic products or less potent product]

Legal requirements with identity, strength, purity and quality

To prevent economic repercussions.

 RATES AND ORDERS OF REACTIONS
RATES
• the speed or velocity of a reaction with which a reactant or
reactants undergoes a change.

• It is determined by the change in the concentration of

the reactants or products as a function of time.

• The rate may be determined by the slowest or rate determining

step.

dc
dt  Rate  kc n
ORDERS OF REACTIONS
the number of concentrations that determine rate.
the way in which the concentration of the reactant influences
the rate.

Law of mass action

 The rate of a reaction is proportional to the molar concentrations of the reactants each raised to
power equal to the number of molecules undergoing reaction.

a A + b B Product Rate α
[A]a .[B]b
Rate = K [A]a .[B]b
Order of reaction = sum of
exponents
Order of A = a and B = b
Then Overall order = a + b
Example:
The reaction of acetic anhydride with ethyl
alcohol to form ethyl acetate and water

(CH3 CO)2 + 2 C2H5OH 2 CH3 CO2 C2H5 +

H2O Rate = K [(CH3 CO)2 O] . [C2H5OH]2

Order for (CH3 CO)2 O is 1st order

Order for [C2H5OH]2 is 2nd order

Overall order of reaction is 3rd Order

 ZERO ORDER REACTIONS

rate is constant and is independent of the concentration of

any of the reactants.

A constant rate of drug release from a dosage form is highly

desirable.
Equation for zero order:
Equation for zero order:

a [A] k Product
(P) Rate = - dc/dt = K
[c]0
- dc/dt
ct =kt dc = -
k dt
 dc    kdt
c0 t0

co = Initial concentration
ct = Concentration at time t
C – C0 = -kt
 C

Units of the rate constant K

c = co – Kt
K = co – c /t
K = Concentration / time
= mole / liter . second
= M. sec-1
Determination of t1/2

Let c = co /2 and t1/2 = t

substitute in equation;
c = co – k t
t1/2 = co / 2K

Note: Rate constant (k) and

t1/2 depend on co

Determination of t0.9

Let c = 0.9 co and t= t0.9

substitute in equation;
c = co –k t
 Examples
• Drug X degrades by a zero-order process with a rate constant
of 0.05 mg ml1 year−1 at room temperature. If a 1%
weight/volume (w/v) solution is prepared and stored at room
temperature:

1. What concentration will remain after 18 months?

2. What is the half-life of the drug?
Answer

1. C0 = 1% w/v = 10 mg/ml; t =18 months = 1.5 year; k0 = 0.05

mg ml−1 year−1
C = C0 – k0t = 10 – (0.05 × 1.5) = 9.93 mg/ml

2. t1/2 = 0.5C0/k0 = (0.5 × 10)/0.05 = 100 years

 FIRST ORDER REACTION

The most common pharmaceutical reactions

e.g; drug absorption & drug
degradation

The reaction rate of change is proportional to drug

concentration.
- dc/dt = kc1 = kc
dc/c = kdt

t t
dc

c0 c
 k  t 0
dt
ln c  ln co  kt
kt
log c  log c0 
2.303
 lnco
C = co e
–kt

Difficult to determine slope

lnc = lnco – kt
Slope = c1 – c2 / t1 – t2
C Lnc
Slope = -k

t t
Log co Log c = log co – kt / 2.303
Slope = c1 – c2 / t1 – t2
Slope = -k / 2.303
Logc

Or use semi log paper

t
Determination of t1/2

Let t = t1/2 and C = C0 /2

substitute in ln C = ln C0 – Kt
t1/2 = ln 2/ K = 0.693 / K t1/2 = 0.693 / K

K units = 0.693 / t1/2 = time-1

Determination of t0.9

Let t = t0.9 c =

0.9 Co substitute in
t0.9 = 0.105 / K and K = 0.105/ t0.9 t0.9 = 0.105 / K
ln c = ln

co – Kt
 Examples
1 Ten (10) ml aqueous solutions of drug A (10% w/v) and drug B
(25% w/v) are stored in two identical test tubes under
identical storage conditions at 37°C for 3 months. If both drugs
degrade by first-order, which drug will retain the highest
percentage of initial concentration?
(a) Drug A
(b) Drug B
(c) They will be the same.

2. The concentration of drug X in aqueous solution drops by 10%

per month when stored at room temperature. If the
degradation occurs by first order, what concentration will
remain if a 5 mg/ml solution of the drug is stored under the
same conditions for 3 months?
3. A 5 gm/100 ml solution of drug X is stored in a closed test tube
at 25°C. If the rate of degradation of the drug is 0.05 day−1,
calculate the time required for the initial concentration to
drop to (a) 50% (half-life) and (b) 90% (shelf-life) of its initial
value.

4. A 5 gm/100 ml solution of drug X is stored in a closed test tube

at 25°C. If the rate of degradation of the drug is 0.05 day−1,
calculate the time for the drug concentration to degrade to
2.5 mg/ml.
 PSEUDO ORDER REACTIONS
• For some reactions, the rate of the reaction may be
independent of the concentration of one or more of the
reacting species over a wide range of reactions.

• These may occur under the following conditions:

One or more of the reactants enters into the rate equation
in great excess compared to others;
One of the reactant is catalyst;
One or more of the reactants is constantly
replenished during the course of reaction
 SECOND ORDER REACTION
Rate depends on the product of two concentration terms.

When you have two components reacting with each other or

one component reacting with itself.

Example: 2HI = H2 + I2 , here the reaction is not simply a

matter of an HI molecule falling apart, but relies on the
collision of two HI molecules.

The rate of reaction from the law of mass action is given by:

Rate = dc/dt = k[HI][HI] = k[HI]2

2nd Order reaction
dc/dt = -kc2

dc/c2 = -kdt
t
c
dc  k dt
c 2

c0 t 0

1 1

 kt c
c0
 2nd order graph

Units of K:

1/C = 1/Co + Kt
K = (1/C - 1/Co) / t
K = M-1. sec -1

i.e, K is dependent on initial drug concentration.

Derive equation for t1/2 and shelf life
Half life: t1/2 = 1 / KCo
Shelf life: t0.9 = 0.11 / KCo
 DETERMINATION OF ORDER AND RATE CONSTANTS

1. Substitution method [data plotting method]

• Data accumulated in experimental kinetic study may
be substituted in the integrated form of the equation that
describes the various reaction orders and observing
which plot is a straight line.
• Accordingly, plot of:
 Concentration against time …….. zero order
reaction [if straight line]
 ln concentration against time ……. First order reaction [if
straight line]
 1/concentration against time …….. second order reaction
[if straight line].
2.Half-life method
• This method is based on the relationship between the initial
concentration of the reactant, the half life, and the reaction
order.

• For zero-order reactions, t1/2 increases with increasing

concentration, whereas for first-order reactions, t1/2 does not
change with change in concentration
 DEGRADATIVE PATHWAYS
Degradation of active drug leads to lowering of quantity of the therapeutic
agent in the dosage form.
It may not be extensive , a toxic product formation may take place due to
decomposition instability of drug product can lead to a decrease in its
bioavailability .
 Changes in physical appearance of given dosage form may take place.
 Degradation may increase or may decrease the potency of drug.
Sometimes active drug may retain its potency , but excipients like –
antimicrobial , preservatives , solubilizers , emulsifying or suspending agent
may degrade , lead to compromising the integrity of drug product.
 EXAMPLE :
Drugs like 5-fluorouracil , carbamazipine , digioxin and theophylline
have narrow therapeutic indices these needs to be carefullytreated in patient
so that plasma levels are neither too high as to be toxic nor too
low as to be ineffective
 The antimicrobial chloroquine can produce toxic reactions that are attribute
to the photochemical degradation of the substance.
 DEGRADATION MAY BE OF TWO TYPES
 PHYSICAL DEGRADATION
 CHEMICAL DEGRADATION
1. OXIDATION
2. DECARBOXYLATIO
3. N PHOTOLYSIS
4. RACEMIZATION
5. HYDROLYSIS
 PHYSICAL DEGRADATION
The physical stability properties includes appearance, palatability ,uniformity
,dissolution and suspend ability are retained . Maintained throughout the shelf
life of the drug.
It includes following :
 Loss of water
 loss of volatile oil
 Water Absorbance
 Polymorphism
 Color change
Physical degradation includes following :
LOSS OF VOLATILE CONTENT: Volatile compounds used such as
Alcohol ether , camphor oils , etc . Try to escape from the formulation leads
to degradation of formulation.
Example : nitroglycerine from drugs evaporate.
LOSS OF WATER : Water loss from liquid preparation (o/w emulsion)
leads to changes in stability .It causes crystallization of drug product .which
may lead to increase in potency , and decrease in weight.
Example : water evaporates from Na2SO4 .BORAX.
WATER ABSORBANCE : pharmaceutical formulations which are
hygroscopic in nature absorb the water from its external environment leads
to degradation .
Example : gelatin capsule , deliquescent salts like –Cacl3 , Potassium
citrate.
POLYMORPHISM: A stable crystal form is effected (it may loosen) leads
to the formation of polymorph and cause instability in formulation. This
may lead to alteration in solubility , dissolution of drug
COLOR CHANGE: Loss or development of color may occur . (due to
change in PH , use of reducing agent , exposure to light )
 Physical Stability
Physical stability implies that:
 The formulation is totally unchanged throughout its shelf
life and has not suffered any changes by way of appearance,
organoleptic properties, hardness, brittleness, particle size
etc.

 It is significant as it affects:
1.pharmaceutical elegance
2.drug content uniformity
3.drug release rate.
 Physical Stability
Formulation Likely physical Effects
instability problems

Oral solutions 1- Loss of flavor Change in smell

2- Change in taste or feel or taste
3Presence of off
flavors due to
interaction with plastic
bottle
4Loss of dye
5- Precipitation
6- discoloration
 Formulation Likely physical Effects
instability problems

Parenteral 1. Discoloration due to Change in

photo chemical appearance and
solutions
reaction or oxidation bio-availability
in
2. Presence of precipitate
due to interaction
with container or
stopper
3. Presence of “whiskers”
4. Clouds due to:
(i) Chemical changes
(ii) The original
preparation of a
supersaturated
solution
 Physical stability
Formulation Likely physical Effects
instability
problems
Suspensions 1 settling 1-Loss of drug
2 caking content
uniformity in
different doses
3 crystal from the bottle
growth
2- loss of
elegance.
 Physical stability
Formulation Likely physical Effects
instability

Emulsions 1 problems
Creaming 1- Loss of drug

2 coalescence content
different
uniformitydoses
in
from the bottle

2- loss of
elegance
Physical stability
 Physical stability
Formulation Likely physical Effects
instability

Semisolids problems
1. Changes in: 1Loss of drug
(Ointments and a) Particle size content
suppositories) b) Consistency uniformity

2loss of
2.Caking or
coalescence elegance

3change in
 Physical stability
Formulation Likely physical Effects
instability problems

Tablets Change in: Change in

a) Disintegration drug release
time
b) Dissolution
profile
c) tfardness
d) Appearance (soft
 CHEMICAL DEGRADATION

Chemical degradation of a dosage form occurs through several pathways like –

hydrolysis , oxidation , decarboxylation , photolysis , racemization. which
may lead to lowering of therapeutic agent in the dosage form, formation of
toxic product , decreased bioavailability etc.
1. HYDROLYSIS
 Most important in systems containing water such as emulsion , suspension ,
solutions , etc.
 Also for drugs which are affected by moisture (water vapor)
from
atmosphere.
 It is usually catalysed by hydrogen ion(acid) or hydroxyl ion(base).
 In this active drug is decomposed with solvent.
 Usually solvent is water some time reaction may involve
pharmaceutical
co solvents such as ethyl alcohol or poly ethylene glycol
 Main classes of drugs that undergo hydrolysis are the ESTERS
Cont…
 ESTER HYDROLYSIS involve acyl – acid cleavage.
 Example of drugs: aspirin ,atropine , physostigmine , procaine..

 R .COOR (ester) + H2O  RCOOH (acid) + HOR(alcohol)

 AMIDE HYDROLYSIS is more stable than ester , susceptible to specific and
general acid base hydrolysis. It involves cleavage of amide linkage to give an
amine instead of alcohol as in case of esters.
 Example of drugs : chloramphenicol , barbiturates .

 RCONHR(amide) + H2 O  RCOOH + NH2

R(AMINE)
 Some functional groups subject to
Hydrolysis
Drug type Examples
Esters Aspirin , alkaloids
Dexmethasne
sodium phosphate

Lactones Nitroglycerin
Pilocarpine
Spironolactone
Amides Chloramphenicol

Imides Glutethimide
Malonic ureas Barbiturates
PROTECTION AGAINST OXIDATION

 Avoiding contact with moisture at time of manufacture.

 Packaging in suitable moisture resistant packs such as strip packs and
storage in
controlled humidity and temperature.
 In liquid dosage form since , hydrolysis is acid or base catalyzed , an optimum
PH for max stability should be selected and the formulation should be stabilized
at this PH by inclusion of proper buffering agents.
 Hydrolysis of certain drugs such as benzocaine and procaine can be decreased
by the addition of specific complexing agent like caffeine to the drug solutions .
 Hydrolysis susceptible drugs such as penicillin and derivatives can be prevented
by formulating them in the dry powder form for reconstitution or dispersible
tablets instead of a liquid dosage form such as solutions or suspensions.
2. OXIDATION

 Oxidation is controlled by environment i.e, light ,trace elements , oxygen and

oxidizing agent .
 Occurs when exposed to atmospheric oxygen.
 Either the addition of oxygen or removal of hydrogen .
 Oxidation is the loss of electrons while reduction is the gain of electrons.

AUTOXIDATION
 The reaction between the compounds and molecular is required
for
oxygen initiating the chain reaction is called autoxidation .
 Free radicals produced during initial reaction are highly reactive and further
catalyze the reaction produced additional free radicals and causing a chain
reaction.
 Heavy metals such as copper , iron , cobalt , and nickel have been known to
catalyze the oxidative degradation .Heat and light further influence the kinetics
of oxidative degradation processes.
STEPS INVOLVED OXIDATION REACTION
 INITIATION : Formation of free radicals is taken place .
R--H  R’ + [H’}
 PROPOGATION : here the free radical is regenerated and react with more oxygen .
R’ + O2  R’—O2
R’O2 + RH  ROOH + R’
 HYDROPEROXIDE DECOMPOSITION
ROOH  RO’ + OH’
 TERMINATION : free radicals react with each other resulting in inactive products.
R’--O2 + X  Inactive product
RO2 + RO2  Inactive product

EXAMPLE OF DRUGS DECOMPOSED BY OXIDATION PATHWAYS

Archis oil , clove oil , ethyl oleate ,Heparin , Ascorbic acid , Morphine ,Vitamin A , Vitamin
B12 , etc.
 PROTECTION AGAINST OXIDATION
USE OF ANTIOXIDANTS : antioxidants are Mainly of 3 types :

1. The first group probably inhibits the oxidation by reacting with free radicals.
Example – tocopheral , butylated hydroxyl anisole (BHA) , butylated hydroxyl
toluene's (BHT). Concentration 0.001 – 0.1%.
2. The second group comprising the reducing agents , have a lower redox potential
than the drug or other substance that they should protect and are therefore more
readily oxidized.
Example –ascorbic acid and iso ascorbic acid , potassium or sodium salts of
metabisulfite.
3. The third group, little antioxidant effect themselelf but enhance the action of
true antioxidant .example
Example -- Citric acid , tartaric acid , disodium edetate and lecithin .

USE OF CHELATING AGENT

when heavy metals catalyze oxidation .
Example -- EDTA , citric acid , tartaric acid form complexes.
3. PHOTOLYSIS
 Exposure to light cause substantial degradation of drug molecule.
• When molecules are exposed to electromagnetic radiation they absorb light
(photons) at characteristic wavelength which cause increase in energy which
can :
 Cause decomposition.
 Retained or transferred.
 Be converted to heat .
 Result in light at a new wavelength (fluorescence ,
emission
phosphorescence).
• Natural sun light lies in wavelength range (290– 780nm) of which only
higher energy (UV) range (290 --320) cause photo degradation of drugs.
 `
 Example of phototoxic drugs:

Furosemide , acetazolamide , cynocobalamine .

 EXAMPLE

Sodium nitropruside in aqueous solution (which is administered by IV

infusion
for management of acute hypertension ).
 If protected from light it is stable to at least 1yr.
 If exposed to normal room light it has a shelf life of 4 hrs.

PROTECTION
 Use of amber colored bottles .
 Storing the product in dark , packaging in cartons also act as physical barrier
to
light.
 Accelerated Stability
Accelerated Stability Studies
Studies
Stability study to predict the shelf life of the product, by accelerating
the rate of decomposition, preferably by increasing the temperature of
reaction conditions.
With the advancement in branch of kinetics, shelf life of a dosage form
can be predicted within months based on accelerated stability reports

Preparations are subjected to high stresses during stability testing.

Common high stresses include :
 Temperature
 Humidity
 Light
It is defined as the time required for the concentration of the reactant to reduce
to 90% of its initial concentration .Represented as t90 and the units of time /conc.
t90 = (a-0.9a) = 0.1 a
ko ko
Where , a = initial concentration .
ko = specific rate constant for zero order reaction.

(the time from the date of manufacture and packaging of the formulation until its
chemical or therapeutic activity is maintained to a predetermined level of labeled
potency and ,
its physical characteristic have not changed appreciably or deleteriously ).
Arrhenius equation
It explains the effect of temperature on rate of a reaction.
According to Arrhenius, for every 10º rise in temperature, the speed
of reaction increases about 2-3 times.

k=Ae -Ea / RT

Ideal gas constant

Arrhenius factor
Energy of activation

Arrhenius factor is the frequency of molecular collisions occuring between

the molecules.

Log k = log A – Ea / 2.303 RT

 Estimation of k value

 The reaction is conducted at several temperatures.

 Concentration of reactants is determined.
 Appropriate graphs are drawn for the kinetic data.
 Data is processed for all the orders.
 The order of the reaction is identified.
 From the slopes of the lines, k values are calculated for all
temperatures.
 Estimation of energy of activation

Activation energy is the minimum energy that a molecule should

possess so that the molecular collisions produce the
product.

 A graph can be drawn by taking log k on y-axis and reciprocal

temperature (1/T) on x-axis.
 A straight line is obtained, the slope of the line is negative and the
magnitude is Ea / 2.303 R.
 The intercept corresponds to log A
 All the constants in the Arrhenius equation can be obtained from
the
graph.
 Steps involved in prediction of shelf
life
 The Preparation is stored at different elevated temperatures, to
accelerate the degradation
 Samples are withdrawn at different time intervals
 The Order of the reaction is determined by plotting the appropriate
function of concentration against time and linear relationship is
determined
 Straight line in a graph permits the estimation of k value from the
slope
 Similarly graphs are drawn for different elevated temperatures.
 K value for each temperature are calculated.
 By using Arrhenius relationship, Log k values are plotted against
reciprocal of absolute temperature, energy of activation can
 Extrapolate the straight line to room temperature (k25) or
refrigerated temperature and read the log k value on y-axis.
 Substitute the k value in the appropriate equation to get the shelf
life of the product.
Arrhenius plot for predicting the rate constant at ambient
temperature(25ºC).
Stability Dosage form Storage condition Storage period
investigation
Storage in open
container until
Organoleptic and Solid equilibrium is reached at 1-2 weeks
physicochemical 25ºC/60%,30ºC/70%,
stability 40ºC/75%
Semisolid 4 weeks
5ºC 4 weeks
≥ - 10ºC
2 weeks
5ºC -40ºC Temperature
cycle within 24 hrs 3 months
Liquid 40ºC(content uniformity)

5ºC
4 Weeks
≥ -10ºC 4 weeks
Photostability
All Xenon lamp
48 hrs
Chemical stability
40ºC, 50ºC, 60ºC, 70ºC 3 months
Solid
Semisolid 30ºC, 40ºC, 50ºC 3 months
40ºC, 50ºC, 60ºC, 70ºC
Liquid 3 months
LONG TERM STABILITY STUDIES :
According to WHO, long term stability testing during and beyond expected shelf life
under storage conditions in the intended market.
RECOMMENDED CONDITIONS FOR LONG TERM STABILITY
STORAGE CONDITIONS
TEMPERATURE (‘C) RELATIVE HUMIDITY% MINIMUM TIME
25’C+/- 2’C 60 +/- 5% 12 MONTHS
30’C +/- 2’C 30+/- 5% 6 MONTHS

ACCELERATED STABILITY STUDIES:

STORAGE CONDITIONS
TEMPERATURE (‘C) RELATIVE HUMIDITY% MINIMUM TIME
40’C +/- 2’C 75 +/-5% 6 MONTHS

In , general the accelerated stability conditions must be at least 15’C above the
actual storage temperature and appropriate relative humidity . Substances and
drugs products intended to be stored in a refrigerator . the accelerated stability
studies should be carried out at 25+/-2’c and 60+/-5% relative humidity.
Testing Frequency:
For Long term testing, during first year sampling should be done
every
three months, during second year, sampling should be done every six
months and after two years, sampling should be done once a year.

Accelerated testing should be done atleast six months and it

suggests
sampling points of 0, 3, 6 months.
Methods Of Accelerated Stability
Testing In Dosage forms

 Freeze Thaw test

 Centrifugal Test

 Shaking test

 Elevated Temperature test

 Accelerated Stability Testing in Emulsions
An emulsion is stored at elevated temperature. This decreases viscosity of the
continuous phase. If the emulsion withstands this stress it is assumed to be
stable at normal conditions of storage.

Centrifugation Method:
Creaming and flocculation are slow processes.
Centrifugation accelerates rate of creaming and flocculation in emulsions.
The emulsion is subjected to different centrifugal speeds and separation of
phases is observed at different time periods.
Bad emulsion separates oil instantly.
Good emulsion does not exhibit detectable separation of oil phase until certain
time period.
Accelerated tests for Suspensions
Cake formation is accelerated by centrifugation.
High speed centrifugation is hence not preferred, low speed centrifugation
is used to study the physical stability.
A Freeze-Thaw cycling technique is one of the stress testing . This cycling
treatment promotes particle growth and has primary importance for
changes in absolute particle size, particle size distribution and
crystal habit.
Accelerated Tests for moisture absorption

In this method, products are placed in an environment of high relative

humidity and controlled temperature.
Their physical and chemical stabilities are assessed.
The results will indicate whether the product is susceptible to moisture
and also whether the container needs to provide a high degree of
protection.
 Limitations

 Stability predictions based on Arrhenius equation are valid only

when the break down depends on temperature.
 The energy of activation obtained in the study should be between
10 to 30 kcal/mole.
 When degradation is due to
Microbial contamination
Photochemical reactions
 When the product looses
its physical integrity at
higher temperatures.
 When the order changes at
elevated temperatures.
 In case of disperse systems,
when temperature is
elevated viscosity is
Describes regarding sampling times ,storage conditions&
specific test parameters for each dosage form.

The FDA & The expert working group of the ICH of technical
requirements for the registration of pharmaceuticals for
human use have published guidelines for conducting the
actual studies.
 ICH Guidelines
• Quality Guidelines “Q” (chemical and pharmaceutical QA)

• Safety Guidelines “S” (in vitro and in vivo pre-clinical studies)

– covering Carcinogenicity Testing, Genotoxicity Testing,
Toxicokinetics and Pharmacokinetics ….. etc.

• Efficacy Guidelines “E” (clinical studies in human subject)

– Covering clinical safety, Dose Response Studies, Good
Clinical Practices, Clinical evaluation …. etc.

• Multidisciplinary Guidelines “M”

– Covering Medical Terminology, Electronic Standards for
Transmission of Regulatory Information …… etc.
– Important for Stability !
» Guideline M4: The Common Technical Document (CTD)
 ICH – Q – Guidelines
Stability Testing Q1
 Stability Testing in Climatic Zone I and II (Q1A)
 Photo stability Testing (Q1B)
 Stability Testing for New Dosage Forms (Q1C)
 Bracketing and Matrixing Designs (Q1D)
 Evaluation of Stability Data (Q1E)
Stability Testing in Climatic Zones III and IV (Q1F)
Validation of Analytical Procedures (Q2)
Impurities (Q3)
 Impurities in New Drug Substances (Q3A)
Impurities in New Drug Products (Q3B)
Pharmacopoeial Harmonization (Q4)
Biotechnological Products (Q5)
Specifications (Q6)
 DEFINITIONS

Shelf life (expiration dating period, conformance period)

Self life is the time period during which a drug product is expected to
remain within the approved specification for use, provided that it is
stored under the conditions defined on the container label.

Re-test period

The period of time during which the drug substance is expected to remain
within its specification and, therefore, can be used in the manufacture of
a given drug product, provided that the drug substance has been stored
under the defined conditions.
Formal stability studies
Long term and accelerated (and intermediate) studies undertaken on primary
and/or commitment batches according to a prescribed stability protocol to
establish or confirm the re-test period of an API or the shelf life of a FPP.
Stress testing – forced degradation (API)
Studies undertaken to elucidate the intrinsic stability of the API. Such testing
is part of the development strategy and is normally carried out under more
severe conditions than those used for accelerated testing.
Stress testing – forced degradation (FPP)
Studies undertaken to assess the effect of severe conditions on the FPP. Such
studies include photostability testing (see ICH Q1B) and compatibility testing
on APIs with each other in FDCs and API(s) with excipients during
formulation development.
 Primary batch (called also exhibit batch)
A batch of an API or FPP used in a formal stability study, from which stability
data are submitted in a registration application for the purpose of establishing a
re-test period or shelf life, respectively. A primary batch of an API should be at
least a pilot scale batch. For a FPP, two of the three batches should be at least pilot
scale batch, and the third batch a production batch.
 Commitment batches
Production batches of a drug substance or drug product for which the stability
studies are initiated or completed post approval through a commitment made in
the registration application.
 Pilot (scale) batch
A batch of an API or FPP manufactured by a procedure fully representative of
and simulating that to be applied to a full production scale batch. (For solid oral
dosage forms, a pilot scale is generally, at a minimum, one-tenth that of a full
production scale or 100,000 tablets or capsules, whichever is the larger.)
 Production (scale) batch
A batch of an API or FPP manufactured at production scale by using production
equipment in a production facility as specified in the application.
 Specification - Release

The combination of physical, chemical, biological, and microbiological tests

and acceptance criteria that determine the suitability of a drug product at the
time of its release.

 Specification - Shelf life

The combination of physical, chemical, biological, and microbiological

tests and acceptance criteria that determine the suitability of an API
throughout its re-test period, or that anFPP should meet throughout its
shelf life.

 Mass balance

The process of adding together the assay value and levels of degradation
products to see how closely these add up to 100% of the initial value, with
due consideration of the margin of analytical error.
 WORLDWIDE ZONES / TEMPERATURE
AND HUMIDITY CONDITIONS

Zone Mean kinetic Yearly average

temperature humidity (%RH)

Zone I ( Moderate) 21 ̊C 45

Zone II (Mediterranean) 25 ̊C 60

Zone III (Hot, dry) 30 ̊C 35

Zone IV (Very hot, moist) 30̊ C 70

 COUNTRIES AND ZONES

Regions Zone I &II Zone III&IV

EUROPE All countries

AMERICA Argentina, Bolivia, Canada, Brazil, Columbia, Cuba,

Mexico, US Jamaica
ASIA Afghanistan, China, Iran, Bahrain , Hong Kong, India,
Nepal, Turkey, Japan Oman , Pakistan, Srilanka,
UAE
AFRICA Egypt, Algeria, South Angola, Benin, Congo,
Africa, Libya Uganda, Sudan, Somalia,
Senegal
 STORAGE CONDITIONS FOR STABILITY STUDY
API/DRUG SUBSTANCES TO BE STOTRED AT AMBIENT TEMPERATURES
Minimum time period covered
Study Storage condition by data at submission
Long term 25°C ± 2°C / 60% ± 5% r.h or 12 months
30°C ± 2°C / 65% ± 5% r.h.
Intermediate 30°C ± 2°C / 65% ± 5% r.h. 6 months

Accelerated 40°C ± 2°C / 75% ± 5% r.h. 6 months

STORAGE IN A REFRIGERATOR
Study Storage condition Minimum time period covered
by data at submission
Long term 5°C ± 3°C 12 months
Accelerated 25°C ± 2°C / 60% ± 5% r.h. 6 months

STORAGE IN FREEZER

Study Storage condition Minimum time period

covered by data at
submission
Long term -20°C ± 5°C 12 months
 DRUG PRODUCTS - PACKAGED IN
SEMI-PERMEABLE CONTAINERS

Study Storage condition Minimum time period

covered by data at
submission

Long term 25°C ± 2°C / 40% ± 5% r.h. or 12 months

30°C ± 2°C / 35% ± 5% r.h.

Intermediate 30°C ± 2°C / 65% ± 5% r.h. 6 months

Accelerated 30°C ± 2°C / 65% ± 5% r.h. 6 months

The Stability Chambers are designed for an operating range of 4°C to
70°C Temperature only, 5°C to 60°C Temperature with Humidity.
These units employ a programmable controller to control the
temperature, defrost and humidity settings. The cabinets use an
evaporator coil, located on top of the cabinet as the heat-removing
source. Through the refrigeration process, heat is captured in the
evaporator, transferred to the condensing unit on top of the cabinet,
and expelled to the surrounding outside air. It is extremely important
to allow a four-inch clearance on the top, rear, and sides of the unit
for the refrigeration process to function properly.
STABILITY CABINETS:
STABILITY CABINETS:
 Packaging And Stability
1.Glass
 Glass is resistant to chemical and

physical change and is the most commonly used

material.

Limitations Overcome
1. Its alkaline surface use of Borosilicate glass
2. Ions may precipitate the use of buffers
insoluble crystals from the
glass
3- Permits the transmission Amber coloured glass
of light which may accelerate
decomposition.
 Packing and Stability
2.PLASTICS
The problems with plastic are:
 Migration of the drug through the into
plastic the
environment.
 Transfer of environmental moisture, oxygen, and
other elements into the pharmaceutical product.
 Leaching of container ingredients into the drug.

Adsorption of the active drug or

excipients by the plastic.

 Packing and Stability
3.Metals
 Various alloys and aluminum 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.
 Packing and Stability

Rubber
 Rubber also has the problems of extraction of
drug ingredients and leaching of container ingredients.

 The pretreatment of rubber vial stoppers and closures with

water and steam reduces potential leaching.
 Physical stability
Formulation Likely physical Effects
instability
problems
Capsules Change in: Change in drug
a) Appearance release

b) Dissolution
c) Strength
 Types of Stability Studies
1.Long-Term (Real-Time) Stability Testing
 Stability evaluation of the physical, chemical, biological
and microbiological characteristics of a drug product

 duration of the shelf life

 Packaging materials permeable to water vapor result in a
falsification of the results for semisolid and liquid dosage forms
if varying degrees of weight loss occur that leads to differences in
the active ingredient concentration or ion strength.

 The use of inert standard packaging materials that are impermeable

to water vapor is important precondition for stress tests that are
evaluated in terms of reaction kinetics, and on the results on which
stability predictions are to be tested.
 Solid dosage forms: 50-mL glass container with twist-off
closure polypropylene tube

 Semisolid dosage forms: Standard tube, small volumetric flask,

Aluminum tube, inert internal lacquering

 Liquid dosage forms: 25mL volumetric flask with ground-glass

stopper

 However, furture investigations for the selection of the final

packaging are necessary.
 On the basis of the results of the stress tests for solid dosage forms,
the sensitivity to moisture can be determined and suitable
packaging materials can be selected.

 As a rule, no interactions are to be expected.

 If the final packaging material has been selected and

samples packed in the final packaging material are available,
the investigation of photostability should be performed.

 Photostability :The samples with and without container are

irradiated with a Xenon lamp for 24 hours.
 Packaging: Aluminum tube internally lacquered, plastic tubes.

 Problems: Corrosion , permeation, sorption.

 Tests packaging material – dosage form:

To test for corrosion ,the filled metal tubes are stored horizontally
upright and inverted at 400C, for 3 months and are then
investigated.

 To test for permeation and sorption the filled plastic tubes

are stored for 3 months at 500C, 400C, 300C/70%.

 If the final packaging material has been selected, the investigations

on the photostability are performed.
 Packaging ampoule, injection vial with rubber stopper, glass bottle
or plastic bottle with screw closure.

 Problems: leakage.

 To test for permeation, and leakage, the finale formulation

solution is filled in the container, and for desorption placebo
solution is used.

 The samples are stored vertically and inverted under 500C, 400C,
300C/70% for up to 12 weeks.

 Tested intervals: 0, 1, 2, 3 months.

 If the final packaging material has been selected the investigations

on the photostability are performed.
TABLE
T

 Stable tablets retain their original size ,shape , weight ,roughness ,colour
variation ,
cracking under normal handling and storage conditions throughout their shelf life.
• FRIABILITY TEST : studies revel the physical instability if any in tablet.
Maximum weight loss should not be more than 1%.

• HARDNESS TEST : shows resistance to crushing.

• COLOR STABILITY : by colorimeter , reflectometer with heat , sunlight and intense

artificial light.

 Uniformity of weight , odor , texture , drug and moisture content , humidity

effects are also Studied during a tablet test.
 GELATINE CAPSULE
Gelatin capsules are found to be stable in dry
conditions but they rapidly reach equilibrium with
the atmospheric conditions under they are stored.
This shows gelatin capsules are largely effected by
temperature and humidity and susceptibility to
microbial degradation .
soft gelatin capsule have Relative Humidity 20 to
30% at 21 to 24’C.
 hard gelatin capsule contain 13 to 16% moisture.
Humidity - capsule shell softens and becomes
sticky.
Dried- capsule shell becomes brittle and crack.
Hard gelatin capsule are tested for Brittleness ,
dissolution , water content and level of microbial
contamination.
 EMULSIONS
Tested for phase separation , PH , viscosity , level of microbial
contamination , and distribution of dispersed globules.
 ORAL SOLUTIONS AND SUSPENSIONS
Formation of precipitate , clarity for solutions , PH , viscosity ,
microbial contamination.
Additionally for suspensions , redispersibility , rheological
properties ,mean size and distribution of particles should be
considered .
 NASAL SPRAYS : solution and suspensions
Clarity (for solution) , level of microbial contamination , PH ,
particulate matter , unit spray medication , content uniformity
, droplet and/or particle size distribution , weight loss , pump
delivery.
Microscopic evaluation ,(for suspension) , foreign particulate
matter and extractable/ leachable from components of the
container , closure and pump.
 TOPICAL , OPTHALMIC AND OTIC PREPRATION
Included in this broad category are ointments ,creams ,
lotions
,paste , gel , solutions ,eye drops and cutaneous sprays.
 TOPICAL

preparations should be evaluated for clarity , homogeneity , PH ,

resuspendibility for lotions , consistency , viscosity , particle size
distribution ,level of microbial contamination / sterility and weight
loss

 FOR OPTHALMIC OR OTIC PREPRATION

Should include the following additional attributes : sterility
,particulate matter ,and extractable.

 SUPPOSITORIES

Softening range , dissolution (at 37’C)

 PARENTERALS
Color , clarity (for solutions) , particulate matter , PH, sterility ,
pyogen / endotoxins .
Stability studies for powders for injection solution ,include color
monitoring , reconstitution time and water content ,to be performed at
regular intervals .