A
Seminar
On
MLIBA PHARMACY COLLEGE,BARDOLI.
1
Contents
 Introduction to Validation
 Stages
of qualifications
 Validation
of Autoclave
 Validation
Protocol of Autoclave
 Validation
of Dry Heat Sterilizers And Tunnel
Validation
Validation
may be defined as ” Establishing documented
evidence which provides a high degree of assurance that a
specific process
meeting its
will consistently produce a product
pre-determined specifications and quality
attributes.”
It
has been made mandatory by the regulatory bodies to
prove the safety efficacy, Purity & effectiveness of the drug
product, medical devices & biologics in the marketplace &
health system.
Why Validation of Equipment?
 Equipment validation
is Vital for
Safety
Fewer
interruptions of work
Lower repair costs
Elimination of premature replacement
Less standby equipment
Identification of high maintenance cost
Reduction of variation in results
Greater confidence in the reliability of results
Who should do Equipment
Validation?
The vendor or the user
 The user has the ultimate responsibility for the accuracy of
the analysis results and also for equipment qualification.

DQ should always be done by the user.

While IQ for a small and low cost instrument is usually
done by the user, IQ for large, complex and high cost
instruments should be done by the vendor.

OQ can be done by either the user or the vendor.

PQ should always be done by the user because it is very
application specific, and the vendor may not be familiar
with these. As PQ should be done on a daily basis, this
practically limits this task to the user.
Validation

Part 1. Overview on qualification and validation

Part 2. Qualification of HVAC and water systems

Part 3. Cleaning validation

Part 4. Analytical method validation

Part 5. Computerized system validation

Part 6. Qualification of systems and equipment

Part 7. Non sterile product process validation
Validation
Stages of qualification
Design qualification
Installation qualification
Operational qualification
Performance qualification
Change control
Validation
Defined schedule
Frequency based on
Factors
Results of calibration
maintenance,
verification
Periodic
Requalification
After change
Extent based on
Risk assessment
Part of
Change control procedure
Equipment qualification

Equipment qualification / validation includes
following things:
 Design
qualification (DQ)
 Installation
qualification (IQ)
 Operational
qualification (OQ)
 Performance
qualification (PQ)
Design Qualification (DQ)
"Design qualification (DQ) defines the
functional and operational specifications of
the instrument and details for the conscious
decisions in the selection of the supplier".
 List below recommends steps that should be
considered for inclusion in a design
qualification.
Description of the analysis problem
Description of the intended use of the
equipment
Description of the intended environment

Preliminary selection of the functional and
performance specifications
Preliminary selection of the supplier
Instrument tests (if the technique is new)
Final selection of the equipment
Final selection of the supplier and equipment
Development and documentation of final
functional and operational specifications
Installation Qualification(IQ)


“Installation qualification establishes that the
instrument is received as designed and specified,
that it is properly installed in the selected
environment, and that this environment is suitable
for the operation and use of the instrument.”
The qualification involves the coordinated efforts
of –
 The
vendor
 The
operating department
 The
project team (which provide input into the
purchase, installation, operation and maintenance
of the equipment).
Operational Qualification (OQ)
 "Operational
qualification (OQ) is the process
of demonstrating that an instrument will
function according to its operational
specification in the selected environment."
 The
proper operation of equipment is verified
by performing the test functions specified in
the protocol.
 A conclusion
is drawn regarding the operation
of equipment after the test functions are
checked and all data has been analyzed.
 Following
are the contents of equipment
operation qualification
1.Application S.O.P’s
2.Utilization List
3.Process Description
4.Test Instrument Utilized To Conduct Test
5.Test Instrument Calibration
6.Critical Parameters
7.Test Function (List)
8.Test Function Summaries
Performance Qualification(PQ)
 "Performance
Qualification (PQ) is the
process of demonstrating that an instrument
consistently performs according to a
specification appropriate for its routine use ".
 PQ should always be performed under
conditions that are similar to routine sample
analysis.
 PQ should be performed on a daily basis or
whenever the equipment is being used.
 In practice, PQ can mean system suitability
testing, where critical key system performance
characteristics are measured and compared
with documented.
A. Introduction
Sterile products have several unique dosage
form properties, such as
 Freedom from micro-organisms,
 Freedom from pyrogens,
 Freedom from particulates,
 Extremely high standards of purity and
quality;
 However, the ultimate goal in the manufacture
of a sterile product is absolute absence of
microbial contamination.

Introduction(Con..)

Three principles are involved in the validation
process for sterile product.
1. To build sterility into a product
2. To demonstrate to a certain maximum level of
probability that the processing and sterilization
methods have established sterility to all units of a
product batch
3. To provide greater assurance and support of the
results of the end product sterility test
D value
“It is time required for a 90% reduction in
microbial population. Quantitative expression
of rate of killing of micro organism.”
 In other words, the D value will be affected by
 The type of microorganism used as BI,
 The
formulation
components
and
characteristics
 The surface on which the micro-organism is
exposed
 The temperature, gas concentration, or
radiation dose of sterilization process.

D value found by 2 methods,
1) Survivor curve method (log number of surviving
organism versus time/gas concentration/radiation dose)
2) Fraction negative method

Z value
 Used exclusively in validation of heat sterilization
process. Z value is reciprocal of slope of plot of log D
verses T at which D value is found i.e. increase in
temperature required to reduce D value of organism by
90 % (1 log reduction)
F value
 Used exclusively in validation of heat sterilization
process. It is time in min required to kill all spores in
suspension at 121oC. Measures equivalent time
Methods of Sterilization of Products
1.Heat

Moist heat (autoclave)

Dry heat oven or tunnel
2.Gas

Ethylene oxide

Peracetic acid

Vapor phase hydrogen peroxide

Chlorine dioxide
3.Radiation

Gamma

Beta

Ultraviolet
B. Qualification and Calibration
1)
Mechanically Checking, Upgrading, and Qualifying the
Sterilizer Unit
 The
main concern with steam sterilization is the complete
removal of air from the chamber and replacement with
saturated steam.
 Autoclaves
can also involve air–steam mixtures for
Sterilizing flexible packaging systems and syringes.
 When
autoclave system is used, the unit must be installed
properly and all operations qualified through installation
qualification and operation qualification (IQ/OQ).
2) Selection and Calibration of Thermocouples

Thermocouples must be durable for repeated use as
temperature indicators in steam sterilization validation and
monitoring.

Copper constantan wires coated with Teflon are a popular
choice as thermocouple monitors.

Accuracy of thermocouples should be 0.5°C. Temperature
accuracy is especially important in steam sterilization
validation.

Thermocouple accuracy is determined using National
Bureau of Standards (NBS).
3) Selection and Calibration of BI
Sr.
No
Sterilization process
Biological Indicator(BI)
1.
Autoclave
B. steriothermophillus spores
B. subtilis var. niger spores
B. subtilis, 5230 spores
B. coagulance spores
Clostridium sporogenes spores
2.
Dry heat
B. subtilis var. niger spores
B. subtilis, 5230 spores
3.
Ethylene Oxide
B. subtilis var. niger spores
4.
Radiation
B. pumilus spores
Micrococcus radiodurans
vegetative cells
C. Heat-Distribution Studies
 Heat-distribution
studies include two phases:
1) Heat distribution in an empty autoclave
chamber
2) Heat distribution in a loaded autoclave
chamber.
 The
trips where the wires are soldered should not
make contact with the autoclave interior walls or
any metal surface.
Cont..
 Heat-distribution
studies may employ thermocouples as
the cool spot in the chamber.
 The
principle is the location of the cool spot and the
effect of the load size and/or configuration on the cool
spot location.
 The
difference in temperature between the coolest spot
and the mean chamber temperature should be not greater
than •
2.5°C .
 Greater
temperature differences may be indicative of
equipment malfunction.
D. Heat-Penetration Studies
 This
is the most critical component of the entire
validation process.
 The
main purpose is to determine the F0 value of
the cold spot inside the commodity.
 The
container cold spot for containers ≥100 ml is
determined using container-mapping studies.
 Thermocouple
probes are inserted within a
container and repeat cycles are run to establish the
point inside the container.
Cont..
 Thermocouples
will be placed both inside and
outside the container at the cool spot location(s),
in the steam exhaust line, and in constanttemperature baths outside the chamber.
 F0
value will be calculated based on the
temperature recorded by the thermocouple inside
the container at the coolest area of the load.
 F0
value will indicate whether the cycle is
adequate or alterations are needed.
Heat-Penetration Studies(Con..)
 Three
critical parameter associated with all
wet heat sterilization Processes:
1. A minimum F value
2. A design F value
3. A sterilization process time
 Any changes in the load size, load
configuration, or container characteristics must
be accompanied;
 To prove that the cool spot location has not
changed or,
 If it has, that it receives the design F0 time
exposure from the sterilization cycle used.
E. Equipment Qualification
 Prior
to the initiation of process, it is important that the
sterilizer be suitably qualified to perform its function.
 Typical
critical requirements that are considered to
affect
the
sterilization
process
(e.g.“quality”
requirements) are:

Accurate temperature and pressure measurement

Air removal to some predefined level of vacuum
 Temperature
chamber.
distribution and uniformity
in
the
 The
qualification of a sterilizer should include the
following :
1.Calibration of temperature and pressure sensors
(traceable to national or international standard)
2.Air removal (usually measured by vacuum level
achieved vs. defined requirement)
3.Demonstration of the sequence of operations,
4.Confirmation of alarms and interlocks
5.Precision of temperature control
6.Temperature distribution and uniformity
F. Microbiological Challenge Studies
 Microbiological
challenges studies are employed to
provide additional necessary assurance that adequate
lethality has been delivered to all parts of the load.
 Calibrated
BIs used as bioburden models providing
data that can be employed to calculate Fo.
 The
microorganisms used to challenge moist heat
sterilization cycles are G. stearothermophilus and
Clostridium sporogenes.
 After
the sterilization cycle is complete, the
inoculated items or spore strips are recovered
and
subjected
to
microbiological
test
procedures.
 Strips
are immersed in a suitable growth
medium (soybean casein digest medium is
typical) and incubated for up to seven days.
G. Sterilizer Filter Evaluation
 Microbial
filters are employed on most parts of
sterilizers to ensure that loads are not contaminated
by air used to vent the chamber as it cools or dries.
 Product
loads
are
protected
from
such
contamination by their primary containers (vials,
bags) and many nonproduct loads are protected by
wraps to provide a microbial barrier.
 For
filters, two issues are of concern:
Sterility and Integrity.
 If
the load will undergo a bioburden cycle, it may
be necessary to sterilize the filter in a separate
phase of the cycle.
 To
ensure that filters will remain functional under
all expected conditions, the integrity tests should
be done following the maximum cycle time and
temperature.
 Triplicate
studies are recommended.
A. Introduction
 Mainly
three types of dry-heat sterilization
systems are utilized in the pharmaceutical
industry today.
I.
Batch Sterilizer Ovens
II.
Tunnel Sterilizers
III.
Microwave Sterilizers

PRINCIPLES OF HEAT TRANSFER
AND
CIRCULATION:
 The
dry heat process must effectively heat the
article, and air surrounding the article, to achieve
sterilization or depyrogenation.
 In
moist heat, the condensation of the steam
sterilizer releases large amounts of heat energy that
serves to heat the items in the sterilizer.
 In
dry heat processes the hot air carries
significantly less heat energy than an equivalent
volume of saturated steam.
Key Process Features to Control Prior to
Validating Dry-Heat Sterilizer
Batch(Oven)
Tunnel Steriliser
Intake air system
Positive pressure to entrance
Exhaust air system
Even distribution of heat
Internal air circulation
Belt speed recorder
Exhaust HEPA filter
HEPA-filtered cooling air
Static pressure gauge
Exhaust HEPA filter
Heater current
Particulate control

The four main mechanism through which Heat
transfer occurs are:
Convection
Circulation
Conduction
Radiation
B. Batch Oven Validation
1. Air balance determination:
 In an empty oven, data are obtained on the flow
rates of both intake and exhaust air.
 Air should be balanced so that positive pressure
is exerted to the nonsterile side when the door is
opened
2. Heat distribution of an empty chamber:
 Thermocouples should be situated according to
a specific predetermined pattern.
 Repeatability of temperature attainment and
identification of the cold spot can be achieved if
the temperature range is •
15°C at all monitored
locations.
3. Heat-penetration studies:

These studies should be designed to determine
the location of the slowest heating point within a
commodity at various locations of a test load in
the sterilizer.
 Thermocouples
are placed in the commodities
located in the areas likely to present the greatest
resistance to reaching the desired temperature.

Minimum
and
maximum
temperatures
as
defined in the process specifications should be
studied.
4. Mechanical repeatability:

During
all
repeatability
these
in
studies,
terms
of
mechanical
air
velocity,
temperature consistency, and reliability and
sensitivity of all the oven and instrumental
controls must be verified.
C. Tunnel Sterilizer Validation
1. Air Balance Determination:
 In
this study items being sterilized are moving exposed to
different air systems (e.g., heating zone and cooling zone).

Air flow must be balanced in order to provide a gradual
decrease in air temperature as items move along the
conveyor.
 In
the absence of a critical balance of air dynamics, either
the items will not be cooled or they will be cooled too
quickly, causing contamination of the entire tunnel area.
2. Heat-Distribution Studies:
 Thermocouples
used in tunnel sterilizer validation
must be sufficiently durable to withstand the extremely
high (≥300°C) temperatures in the heating zone area of
the tunnel.
 Heat-distribution
studies should determine where the
cold spots are located as a function of the width of the
belt and height of the tunnel chamber.
 Peak
temperature readings should remain within •
10°C
across the belt for at least three replicate runs.
3. Heat-Penetration Studies:
 Prior
to microbial challenge testing of the tunnel
sterilization,
heat-penetration
studies
must
be
completed in order to identify the coolest container in
the entire load.
 Three
to five replicate runs for each commodity size
and every loading configuration should be
done
using 10 to 20 thermocouples distributed throughout
the load.
 Careful
analysis of the temperature data after each
run will be invaluable in the determination of the cool
spot
4. Mechanical Repeatability:
 Tunnel
sterilizers
must
demonstrate
mechanical repeatability in the same manner
as batch ovens.
 Air
velocity, air particulates, temperature
consistency and reliability of all the tunnel
controls (heat zone temperatures, belt speed)
must be proved during the physical validation
studies.
D. Biological Process Validation of Dry
Heat Sterilization Cycles
 If
the dry-heat process is claimed to produce both
sterile and pyrogen-free commodities, validation
studies must be done using both micro-organisms
and microbial endotoxins.
 The
goal is to validate a heating cycle that can
produce a 12-log reduction in the biological
indicator population.
 The
most widely used biological indicators for
dry heat have been spores of B. Subtilis.

Procedures for the validation of a tunnel sterilization:
 The
overkill approach is selected for the validation
study.
 Select
 Run
the type of biological indicator to be used.
a complete cycle using the desired loading
pattern.
 Determine
the number of survivors by plate-counting
or fraction negative Methods.
 Determine
the number of spore log reductions (SLRs)
E. Endotoxin challenge in Dry Heat
Sterilization
 Inoculate
commodity samples with a known
amount of endotoxin. (e.g., 10–100 ng Escherichia
coli lipopolysaccharide)
 Thermocouples
adjacent
to
should be placed in commodities
those
containing
endotoxin
for
temperature monitoring and correlation with LAL
test results.
 Endotoxin
destruction should be ascertained at the
coolest location of the load.
 Several
endotoxin challenge samples should be
done per cycle, and the studies must be adequately
replicated.
 Following
the dry-heat cycle, aseptically transfer
the units containing endotoxin to an aseptic area
for extraction procedures.
F
values required for endotoxin destruction at
various
temperatures
and/or
cycle
time–
temperature variations can be determined using a
Z value of 54°C.
VALIDATION OF TEST
EQUIPMENT
Equipment
required to conduct the IQ, OQ and
PQ are discussed here.
All
temperature equipment employed to
perform the validation studies must be
traceable and calibrated to the International
Temperature Scale

The equipments used for validation testing of dry heat
processes are discussed here:
 Resistance Temperature
Detectors
 Thermocouples
 Data
Loggers
 Wireless Temperature
Logger
 Infrared Thermometer
 Constant Temperature
 Stopwatch
 Voltmeter
or Ammeter
 Optical Tachometer
Baths
INSTALLATION QUALIFICATION
 The
IQ is designed to compare the system against the
manufacturer’s specifications for proper installation.
 All
equipment, utilities, and connections must be
checked
against
the
manufacturer’s
recommendations.
A. Structural:
 Check
dimensions, presence of identification plates,
correct leveling, proper insulation, presence of seals,
and inspect for structural damage.
B. Filters:
 All filters used within the system must be
recorded, such as those used with air (supply, recirculating) or in other utilities (e.g., steam,
water).
 Some HEPA filters may need to be checked
periodically by performing an integrity test or
DOP.
C. Electrical:
 Ensure conformance to National Electrical Code
Standards
D. HVAC:
 Ensure the system provides the RH, temperature,
and pressure differential required.
E. Air Supply:
 Identify source (direct from the HVAC system
or room air), duct size, duct material of
construction, and air classification.
F. Ventilation:
 Check that the ventilation exhaust duct
exhausts to an appropriate area (not to an
aseptic environment), and identify the method
used to prevent back-flow.
G. Door Gaskets:
 Check integrity of gaskets and materials of
construction.
H. Heaters:
 Record the manufacturer’s model number, the
number of heating elements, and the voltage,
amperage, and wattage of the elements for the
heaters.
I. Lubricants:
 Make certain that any lubricants used cannot
contaminate the material being sterilized or
depyrogenated.
J. Blowers:
 The blower must be mechanically sound, the
volute in place and correctly balanced, and that
the blades rotate in the correct direction.
OPERATIONAL QUALIFICATION
A. Temperature Monitors:
 The temperature controllers, recorders, and
sensors on the process equipment must be
calibrated before the unit can be operated
reliably.
B. Cycle Timer:
 The accuracy of the timer must be determined,
so that assurance is provided for cycle length.
C. Door Interlocks:
 If a unit is equipped with double doors, the
interlocks must operate such that the door
leading to the aseptic area cannot be opened.
D. Heaters:
 All of the heating elements must be functional. It
is preferable to have them monitored
continuously with ammeters in order that burnedout elements can be immediately detected.
E. Cooling Coils:
 To enable a faster cool-down cycle, the air is
often circulated across coolant coils.
F. Belts:
 The belt speed is a critical operating parameter in
both continuous hot-air tunnels and flame
sterilizers.
 Recorders for charting the belt speed are
recommended for units with adjustable speed
settings.
G. Particulate Counts:
 Particulate
counts should be checked within
the containers before and after sterilization to
quantitate the particle load.
H. Chamber Leaks:
 The
perimeter of the doors for batch sterilizers
should be checked for air leakage while
operating.
QUALIFICATION TESTING
 Upon
completion of IQ and OQ efforts and
approval of the protocol, testing may begin.
 The
testing will include empty-chamber testing
for:

Heat distribution studies,

Loaded-chamber testing consisting of heat
distribution and heat penetration studies.
1) Component Mapping Studies
 Before conducting the loaded-chamber heat
penetration studies, component mapping
should be conducted.
 The studies help to determine the coolest point
within a specific load and item.
2) Empty-Chamber Testing
 The initial testing is performed on an empty
oven or tunnel to establish the uniformity of
temperature distribution.
 The thermodynamic characteristics of the
empty unit are depicted in a temperature
distribution profile.
3) Loaded-Chamber Studies
 For
validation purposes, the loads tested must be
representative of standard items and quantities.
 Ideally,
each size and type of material should be
tested by penetration studies.
 For
ovens, the time and temperature set points
should be reduced. For tunnels, the temperature set
point should be reduced and the belt speed
increased if possible.
4) Bio-Challenge/Pyro-Challenge Studies

The challenge should demonstrate the lethality
delivered by the cycle with either microorganisms
or endotoxin.

The challenge can be accomplished using
commercial strips or suspensions of B. subtilis
spores for sterilization or E. Coli endotoxin for
depyrogenation.

The concentration of the challenge for overkill
processes must demonstrate adequate sterility
assurance.
QUALIFICATION REPORT
 After
the empty and loaded-chamber studies and biochallenge studies have been completed, the data must
be analyzed to ascertain that all testing requirements
have been achieved.
 The results of the biochallenge studies and F value
computation must demonstrate the required degree of
lethality according to the protocol.
 The following information should be provided in the
process qualification validation report:
1. Protocol achievement
2. Summary of data
3. Deviations
4. Diagram
References
 Berry
I.R., and Nash R.A., ”Pharmaceutical Process
validation” second edition, revised and expanded;
Marcel Dekker series; 83-110.
 Agalloco
J.A,
Carleton
F.A,
”Validation
of
Pharmaceutical Process, Third Edition, 175,223.
 www.fda.gov
 Wood,
R.T; Journal of Parental drug association;
volume 34; 286-294
 Groves,
M.J.; Journal of Parental Technology; 2nd
edition; 432
73
Topic is open for Discussion
74