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MLIBA PHARMACY COLLEGE,BARDOLI.

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A Seminar On

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 will consistently produce a product

meeting its 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 forSafetyFewer interruptions of workLower repair costsElimination of premature replacementLess standby equipmentIdentification of high maintenance costReduction of variation in resultsGreater confidence in the reliability of results

Who should do Equipment Validation?

The vendor or the userThe 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

ValidationStages of qualification

Design qualification

Installation qualification

Operational qualification

Performance qualification Change control

Validation

Requalification

Defined schedule

Frequency based onFactors

Results of calibrationmaintenance,

verification

Periodic

After change

Part ofChange control procedure

Extent based onRisk assessment

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 problemDescription of the intended use of the

equipmentDescription 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. IntroductionSterile products have several unique dosage

form properties, such asFreedom 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 valueUsed 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 valueUsed 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 BISr. No

Sterilization process Biological Indicator(BI)

1. Autoclave B. steriothermophillus sporesB. subtilis var. niger sporesB. subtilis, 5230 sporesB. coagulance sporesClostridium sporogenes spores

2. Dry heat B. subtilis var. niger sporesB. subtilis, 5230 spores

3. Ethylene Oxide B. subtilis var. niger spores

4. Radiation B. pumilus sporesMicrococcus 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 constant-

temperature 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 value2.A design F value3.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 distribution and uniformity in the

chamber.

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 Validation1. 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 these studies, mechanical

repeatability in terms of 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 D. Biological Process Validation of Dry Heat Sterilization CyclesHeat 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 the type of biological indicator to be used.

Run 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 should be placed in commodities

adjacent to 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 Baths

Stopwatch

Voltmeter or Ammeter

Optical Tachometer

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, re-circulating) 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

StandardsD. 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 QUALIFICATIONA. 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 burned-out 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 StudiesBefore 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 TestingThe 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 bio-

challenge 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

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