A Seminar On

MLIBA PHARMACY COLLEGE,BARDOLI.

Contents
Introduction to Validation Stages

of qualifications of Autoclave

Validation

Validation
Validation

Protocol of Autoclave
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 attributes.
It

will consistently produce a product

pre-determined specifications and quality

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
Results of calibration maintenance, verification

Defined schedule
Frequency based on Factors

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) qualification (IQ) qualification (OQ)

Installation Operational

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 vendor operating department project team (which provide input into the

The The 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."
proper operation of equipment is verified by performing the test functions specified in the protocol. is drawn regarding the operation of equipment after the test functions are checked and all data has been analyzed.

The

A conclusion

 Following

are the contents of equipment operation qualification

1.Application S.O.Ps

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 Ethylene oxide Peracetic acid

2.Gas

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 airsteam 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 0.5C. 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 1. Sterilization process Autoclave Biological Indicator(BI) 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 B. subtilis var. niger spores

3.

Ethylene Oxide

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 2.5C .
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 container cold spot for containers 100 ml is

the cold spot inside the commodity.

The

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

the sterilization process (e.g.quality

affect

requirements) are:

Accurate temperature and pressure measurement Air removal to some predefined level of vacuum distribution and uniformity in the

Temperature

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. II.

Batch Sterilizer Ovens 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)
Intake air system Exhaust air system Internal air circulation Exhaust HEPA filter

Tunnel Steriliser
Positive pressure to entrance Even distribution of heat Belt speed recorder HEPA-filtered cooling air

Static pressure gauge

Heater current

Exhaust HEPA filter

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 15C 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 studied.

and

maximum

temperatures

as

defined in the process specifications should be

4. Mechanical repeatability:

During

all

these in

studies, of

mechanical air velocity,

repeatability

terms

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 (300C) 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 10C

across the belt for at least three replicate runs.

3. Heat-Penetration Studies:
Prior

to microbial challenge testing of the tunnel heat-penetration studies must be

sterilization,

completed in order to identify the coolest container in

the entire load.

Three

to five replicate runs for each commodity size done

and every loading configuration should be the load.

Careful

using 10 to 20 thermocouples distributed throughout

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: overkill approach is selected for the validation

The

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., 10100 ng Escherichia

coli lipopolysaccharide)
Thermocouples

should be placed in commodities containing endotoxin for

adjacent

to

those

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 54C.

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 Thermocouples Data

Detectors

Loggers Logger

Wireless Temperature

Infrared Thermometer
Constant Temperature

Baths

Stopwatch
Voltmeter

or Ammeter

Optical Tachometer

INSTALLATION QUALIFICATION
The

IQ is designed to compare the system against the

manufacturers specifications for proper installation.

All

equipment, utilities, and connections must be against the manufacturers

checked

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 manufacturers 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

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Topic is open for Discussion

74