Sterile Product Manufacturing

Introduction To give an overview of the principles involved in the

manufacture of sterile products

The overall objective is to produce product that has a high assurance of sterility (and which meets all other quality parameters)

This presentation: Summarises the general approach Gives a framework for other detailed guides on specific aspects of

sterilisation & sterile manufacturing Illustrates the underlying principles Provides advice and gives recommendations.

To give an overview of the principles involved in the manufacture of sterile products

The overall objective is to produce product that has a high assurance of sterility (and which meets all other quality parameters)

This presentation: Summarises the general approach Gives a framework for other detailed guides on specific aspects of

sterilisation & sterile manufacturing Illustrates the underlying principles Provides advice and gives recommendations.

General Principles of Sterile Manufacturing

Moist Heat Sterilization Dry Heat Sterilization Aseptic Processing Environmental Monitoring Ethylene Oxide Sterilization Sterile Filtration Water systems validation Sterility testing Radiation Sterilization Visual Inspection

Moist Heat Sterilization Dry Heat Sterilization Aseptic Processing Environmental Monitoring Ethylene Oxide Sterilization Sterile Filtration Water systems validation Sterility testing Radiation Sterilization Visual Inspection

Fundamentals

Sterility is the absence of living organisms This is an absolute definition

The probability of achieving sterility depends on the overall process

It is generally accepted that a terminally sterilized product should have a probability of non-sterility of less than 10-6 (i.e., a lower probability than one in a million of having a non-sterile unit)

This is often expressed as an SAL Sterility Assurance Level of 106

This is a worst-case figure (with a challenge more resistant than product bioburden). Real confidence levels are generally very much higher

A figure that has sometimes been quoted for aseptically filled product is probability of non-sterility of less than 10-3. However, this is harder to analyse as contamination does not follow a clear statistical distribution. Potential contamination sources are not randomly distributed.

Sterility is the absence of living organisms This is an absolute definition

The probability of achieving sterility depends on the overall process

It is generally accepted that a terminally sterilized product should have a probability of non-sterility of less than 10-6 (i.e., a lower probability than one in a million of having a non-sterile unit)

This is often expressed as an SAL Sterility Assurance Level of 106

This is a worst-case figure (with a challenge more resistant than product bioburden). Real confidence levels are generally very much higher

A figure that has sometimes been quoted for aseptically filled product is probability of non-sterility of less than 10-3. However, this is harder to analyse as contamination does not follow a clear statistical distribution. Potential contamination sources are not randomly distributed.

Why Validate and Control?

The test for sterility cannot confirm that the whole batch is sterile It is performed on a sample from a batch and has

statistical limitations It can miss contamination if only a proportion of units

are non-sterile It is thus necessary to recognize and understand

every aspect that could lead to loss of sterility assurance

Such conditions should be prevented by the application of carefully designed barriers and/or control measures.

The test for sterility cannot confirm that the whole batch is sterile It is performed on a sample from a batch and has

statistical limitations It can miss contamination if only a proportion of units

are non-sterile It is thus necessary to recognize and understand

every aspect that could lead to loss of sterility assurance

Such conditions should be prevented by the application of carefully designed barriers and/or control measures.

Development Validation and Control

It is important that the product and process are designed to

maximise sterility assurance

Wherever possible, the product should be developed to withstand

sterilization in the final container

Once the product design is defined, a suitable production process

must be developed

This is installed and validated

The process must then be tightly controlled to assure reliability and

consistency.

It is important that the product and process are designed to

maximise sterility assurance

Wherever possible, the product should be developed to withstand

sterilization in the final container

Once the product design is defined, a suitable production process

must be developed

This is installed and validated

The process must then be tightly controlled to assure reliability and

consistency.

Product Design Considerations

Facility Design

Cleaning and disinfection of the Facility

Water

Categories of Water

Gases and Vacuum

Equipment (1)

Equipment (2)

Equipment Sterilization and handlingSterilization must follow a validated procedureAseptic processes designed to minimise aseptic

assembly and interventionUnavoidable aseptic assembly needs clear &

precise proceduresAseptic assembly must be simulated (worst-case)

in media fill simulation trials Sterilization In Place is a good method where

possible must be validated.

Equipment Sterilization and handlingSterilization must follow a validated procedureAseptic processes designed to minimise aseptic

assembly and interventionUnavoidable aseptic assembly needs clear &

precise proceduresAseptic assembly must be simulated (worst-case)

in media fill simulation trials Sterilization In Place is a good method where

possible must be validated.

Personnel

Training - personnel appropriately trained for sterile processing, including assessment and documentation: Basic GMP Fundamentals of microbiology Personal hygiene, health and cleanliness Behaviour and aseptic working techniques Gowning and entry procedures Cleaning and disinfection Sterilization procedures, validation and routine operation Emergency procedures to protect product quality (e.g. loss of HVAC System, loss of

power, equipment interventions etc.) Personnel participating in aseptic processing must have practical

training in aseptic techniques before doing aseptic manipulations They must have participated in a successful media fill run.

Training - personnel appropriately trained for sterile processing, including assessment and documentation: Basic GMP Fundamentals of microbiology Personal hygiene, health and cleanliness Behaviour and aseptic working techniques Gowning and entry procedures Cleaning and disinfection Sterilization procedures, validation and routine operation Emergency procedures to protect product quality (e.g. loss of HVAC System, loss of

power, equipment interventions etc.) Personnel participating in aseptic processing must have practical

training in aseptic techniques before doing aseptic manipulations They must have participated in a successful media fill run.

Gowning and Aseptic Technique

Gowning Personnel must correctly wear appropriate clean room garments Detailed, easily understood, gowning procedure (preferably illustrated)

Aseptic Techniques Personnel in the aseptic manufacturing area, must understand the

principles of aseptic procedures They must only be considered qualified after appropriate training,

working under supervision and demonstration of competence The supervisor should observe technique & correct as necessary All personnel directly involved in aseptic processing must participate in

a media fill at least once per year

Glove disinfection Sterile disinfectants must be available (e.g., alcohol based) Glove disinfection must be reasonably frequent, defined in SOP.

Gowning Personnel must correctly wear appropriate clean room garments Detailed, easily understood, gowning procedure (preferably illustrated)

Aseptic Techniques Personnel in the aseptic manufacturing area, must understand the

principles of aseptic procedures They must only be considered qualified after appropriate training,

working under supervision and demonstration of competence The supervisor should observe technique & correct as necessary All personnel directly involved in aseptic processing must participate in

a media fill at least once per year

Glove disinfection Sterile disinfectants must be available (e.g., alcohol based) Glove disinfection must be reasonably frequent, defined in SOP.

Environmental Monitoring (1)

The scope of environmental monitoring includes:Non-viable particulates,Viable (microbial) countsDifferential pressuresTemperaturesHumiditiesAir flows

The scope of environmental monitoring includes:Non-viable particulates,Viable (microbial) countsDifferential pressuresTemperaturesHumiditiesAir flows

Environmental Monitoring (2)

Monitoring During Room Qualification Operational Qualification (OQ) at rest conditions to verify operation Performance Qualification (PQ) in worst case operational conditions Action levels should meet USP or Euro GMP as applicable Alert levels tight enough to detect deterioration, but not so tight that

they become meaningless due to frequent transgression PQ must cover a sufficient period to establish consistency

Routine Monitoring Ensures area remains satisfactory. Results should be within alert level Results above alert levels need review and perhaps corrective actions Above action levels, must trigger appropriate actions (described in

guide), Results must be assessed for trends so that progressive or sudden

changes in the results may be observed. This should be reviewed regularly.

Monitoring During Room Qualification Operational Qualification (OQ) at rest conditions to verify operation Performance Qualification (PQ) in worst case operational conditions Action levels should meet USP or Euro GMP as applicable Alert levels tight enough to detect deterioration, but not so tight that

they become meaningless due to frequent transgression PQ must cover a sufficient period to establish consistency

Routine Monitoring Ensures area remains satisfactory. Results should be within alert level Results above alert levels need review and perhaps corrective actions Above action levels, must trigger appropriate actions (described in

guide), Results must be assessed for trends so that progressive or sudden

changes in the results may be observed. This should be reviewed regularly.

Environmental Monitoring (3)

Deviation Reports and Failure Investigations The data must be analysed Where necessary further investigations initiated Possible contamination sources to be assessed and, eliminated Outcome and detail must be reported

Recommended Methods for Routine Monitoring Physical measurements of the air supply Physical and microbiological monitoring of the environment Particles (viable and non-viable) in the air Micro-organisms settling out of the air Micro-organisms contaminating surfaces Presence of micro-organisms on the hands and garments

Monitoring Plan Defined monitoring plans: tests, locations, alert/action levels & frequencies May contain details of water, compressed gas clean steam testing A review of environmental data is a requirement for batch release.

Deviation Reports and Failure Investigations The data must be analysed Where necessary further investigations initiated Possible contamination sources to be assessed and, eliminated Outcome and detail must be reported

Recommended Methods for Routine Monitoring Physical measurements of the air supply Physical and microbiological monitoring of the environment Particles (viable and non-viable) in the air Micro-organisms settling out of the air Micro-organisms contaminating surfaces Presence of micro-organisms on the hands and garments

Monitoring Plan Defined monitoring plans: tests, locations, alert/action levels & frequencies May contain details of water, compressed gas clean steam testing A review of environmental data is a requirement for batch release.

Bioburden and Components

Active Ingredients, Excipients, Additives All ingredients should have appropriate biological specifications Any limitations to sterilization must be defined Description of origin (e.g. virological / prion risk)

Materials Used in the Process Where appropriate, determine bioburden (e.g., ion exchange materials)

Primary Packaging Components Container and the closure and cleaning / sterilization to be clearly

specified Steps such as siliconization may need monitoring If cleaning/sterilization is by supplier, same exigencies apply

Container-closure integrity The integrity must be validated Simulate, where appropriate: stress from processing Method appropriate to container/closure system

Active Ingredients, Excipients, Additives All ingredients should have appropriate biological specifications Any limitations to sterilization must be defined Description of origin (e.g. virological / prion risk)

Materials Used in the Process Where appropriate, determine bioburden (e.g., ion exchange materials)

Primary Packaging Components Container and the closure and cleaning / sterilization to be clearly

specified Steps such as siliconization may need monitoring If cleaning/sterilization is by supplier, same exigencies apply

Container-closure integrity The integrity must be validated Simulate, where appropriate: stress from processing Method appropriate to container/closure system

Weighing, Compounding and Sterilization

Weighing and compounding must be carried out in suitably classified rooms Vessels must be cleaned, and sterilized or sanitised as appropriate and

stored dry in a way to prevent microbial contamination

Storage of pre-sterilization intermediates to be controlled & time limited Following aspects to be considered:

Pre-filtration bioburden (filter sterilized material) Pre-sterilization bioburden Appropriate in-process controls

Sterilization of product and product contact materials Selection of a suitable sterilization protocol must be based on SAL Method must also consider the stability of the product Validation always required Change control is vital; even apparently minor change must be assessed

Weighing and compounding must be carried out in suitably classified rooms Vessels must be cleaned, and sterilized or sanitised as appropriate and

stored dry in a way to prevent microbial contamination

Storage of pre-sterilization intermediates to be controlled & time limited Following aspects to be considered:

Pre-filtration bioburden (filter sterilized material) Pre-sterilization bioburden Appropriate in-process controls

Sterilization of product and product contact materials Selection of a suitable sterilization protocol must be based on SAL Method must also consider the stability of the product Validation always required Change control is vital; even apparently minor change must be assessed

Terminal Sterilization

Steam Sterilization By far the most common method for aqueous-based pharmaceuticals Preferred cycle is the Pharm Eur reference cycle is 15 minutes at 121C The sterilization cycle chosen must be compatible with product stability Sterilization parameters clearly defined In conjunction with other controls, the required SAL must be demonstrated Validation to confirm sterilization conditions consistently throughout the load

Sterilization by Ionizing Radiation Common for medical devices, but not for pharmaceuticals. Pharm. Eur. reference condition, 25 KiloGray (kGy), has been widely accepted. Other

conditions may be used if validated and accepted by the regulator Important to consider susceptibility of the product to radiation damage

Dry Heat Sterilization Lower antimicrobial efficacy than moist heat, thus higher temperatures and/or longer

exposures. Pharm Eur reference cycle is 2 hours @ 160C Rarely used for terminal sterilization of pharmaceuticals; in rare cases heat resistant

non-aqueous products may be terminally sterilized.

Steam Sterilization By far the most common method for aqueous-based pharmaceuticals Preferred cycle is the Pharm Eur reference cycle is 15 minutes at 121C The sterilization cycle chosen must be compatible with product stability Sterilization parameters clearly defined In conjunction with other controls, the required SAL must be demonstrated Validation to confirm sterilization conditions consistently throughout the load

Sterilization by Ionizing Radiation Common for medical devices, but not for pharmaceuticals. Pharm. Eur. reference condition, 25 KiloGray (kGy), has been widely accepted. Other

conditions may be used if validated and accepted by the regulator Important to consider susceptibility of the product to radiation damage

Dry Heat Sterilization Lower antimicrobial efficacy than moist heat, thus higher temperatures and/or longer

exposures. Pharm Eur reference cycle is 2 hours @ 160C Rarely used for terminal sterilization of pharmaceuticals; in rare cases heat resistant

non-aqueous products may be terminally sterilized.

Sterilization of Items for Aseptic Fill (1)

Steam Sterilization Widely used, but careful validation needed particularly complex items Broadly similar to terminal steam sterilization, but two aspects are critical

Quality of saturated steam Removal of air and subsequent steam penetration

Sterilization by Ionizing Radiation May be used for temperature sensitive primary packaging or components Used for disposables for sterile areas and sterility testing areas Validation includes dosimetry, - correct, even, irradiation of the items

Dry Heat Sterilization/Depyrogenation Sterilization/ depyrogenation of heat resistant primary packaging materials Pharm Eur notes that temperatures in excess of 220 oC have been frequently used,

the USP suggests 250 15 oC Validation must include endotoxin challenge studies Dry heat may be used to sterilize non-aqueous preparations (e.g. Ointment bases) at

lower temperature/time relationships, without depyrogenation.

Steam Sterilization Widely used, but careful validation needed particularly complex items Broadly similar to terminal steam sterilization, but two aspects are critical

Quality of saturated steam Removal of air and subsequent steam penetration

Sterilization by Ionizing Radiation May be used for temperature sensitive primary packaging or components Used for disposables for sterile areas and sterility testing areas Validation includes dosimetry, - correct, even, irradiation of the items

Dry Heat Sterilization/Depyrogenation Sterilization/ depyrogenation of heat resistant primary packaging materials Pharm Eur notes that temperatures in excess of 220 oC have been frequently used,

the USP suggests 250 15 oC Validation must include endotoxin challenge studies Dry heat may be used to sterilize non-aqueous preparations (e.g. Ointment bases) at

lower temperature/time relationships, without depyrogenation.

Sterilization of Items for Aseptic Fill (2)

Ethylene Oxide Sterilization Quite widely used to sterilize heat labile components European Pharmacopoeia and the European GMP guide indicate

that this method should only be used where there is no suitable alternative

Hazardous - toxic, potentially carcinogenic, flammable, potentially explosive

Generally conducted by specialized contractors There are strict regulatory limits on maximum permissible

product residues Bulk packs for sterilization must be gas permeable, but sealed

against microbial ingress Sterilization must consider packaging, load pattern, gas

penetration (ethylene oxide & water vapour), bulk pack integrity Validation and routine monitoring must include Biological

indicators.

Ethylene Oxide Sterilization Quite widely used to sterilize heat labile components European Pharmacopoeia and the European GMP guide indicate

that this method should only be used where there is no suitable alternative

Hazardous - toxic, potentially carcinogenic, flammable, potentially explosive

Generally conducted by specialized contractors There are strict regulatory limits on maximum permissible

product residues Bulk packs for sterilization must be gas permeable, but sealed

against microbial ingress Sterilization must consider packaging, load pattern, gas

penetration (ethylene oxide & water vapour), bulk pack integrity Validation and routine monitoring must include Biological

indicators.

Sterilization by Filtration (Liquids)

Principle: Contaminating organisms are not killed, but are retained on the filters. Any faults in

the filter structure, may compromise this Validation includes:

Retention of bacterial challenge: B. diminuta at 107 per cm2 This is correlated with an integrity test value

Validation should address: Filter suitability - toxicity, extractables, shedding of particles Adsorption of product Compatibility with product solvents The required filter size and suitability of the filtration equipment Retention of B.diminuta in the actual product under process conditions Parameters for the physical integrity test

Routine Filtration Conducted in line with the validated parameters Check integrity testing, process time, differential pressure, flow rates, sterilization and

reuse of filters.

Principle: Contaminating organisms are not killed, but are retained on the filters. Any faults in

the filter structure, may compromise this Validation includes:

Retention of bacterial challenge: B. diminuta at 107 per cm2 This is correlated with an integrity test value

Validation should address: Filter suitability - toxicity, extractables, shedding of particles Adsorption of product Compatibility with product solvents The required filter size and suitability of the filtration equipment Retention of B.diminuta in the actual product under process conditions Parameters for the physical integrity test

Routine Filtration Conducted in line with the validated parameters Check integrity testing, process time, differential pressure, flow rates, sterilization and

reuse of filters.

Performance Qualification of Aseptic Manufacturing

Based on simulating the risk of contamination in all aseptic operations

For a new process, a minimum of three consecutive satisfactory media filling trials

For aqueous liquid products, simulation trials use a liquid microbiological medium

For solid dosage forms, a powder placebo is used, followed by aseptic reconstitution into a liquid microbiological medium

The following slide gives a general overview....

Based on simulating the risk of contamination in all aseptic operations

For a new process, a minimum of three consecutive satisfactory media filling trials

For aqueous liquid products, simulation trials use a liquid microbiological medium

For solid dosage forms, a powder placebo is used, followed by aseptic reconstitution into a liquid microbiological medium

The following slide gives a general overview....

Aseptic Process Simulation (Media Fill Trial)

Media Fill Trials (MFTs) All process stages simulated as closely as possible Particularly interventions and manual manipulations Must follow routine procedures and include all interventions Regular interventions simulated with the same frequency as actual

process In each case, the worst-case eventuality must be covered Process must be successfully validated before product filling is

permitted Revalidation by media fill must be conducted every half year (each line)

Manufacturing Environment Microbiological monitoring must be performed during the trial

Filling Conditions and Equipment All according to routine operating conditions and at normal times of day Containers must be passed through all stages.

Media Fill Trials (MFTs) All process stages simulated as closely as possible Particularly interventions and manual manipulations Must follow routine procedures and include all interventions Regular interventions simulated with the same frequency as actual

process In each case, the worst-case eventuality must be covered Process must be successfully validated before product filling is

permitted Revalidation by media fill must be conducted every half year (each line)

Manufacturing Environment Microbiological monitoring must be performed during the trial

Filling Conditions and Equipment All according to routine operating conditions and at normal times of day Containers must be passed through all stages.

Thank You

Any Questions

Thank You

Any Questions

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