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STERILE PROCESS OF VALIDATIONSTERILE PROCESS OF VALIDATION
Contents
IntroductionAseptic sterile- ldquo A state of control attained by using an aseptic work area and performing activities in a manner that precludes microbiological contamination of the exposed sterile productrdquo
Process Validation- ldquoProcess validation is establishing documented evidence which demonstrate that the manufacturing process will consistently produce a product meeting its predeterminespecifications and quality Characteristicsrdquo
Drug product container and closure are subject to sterilization separately and then brought together
Validation- is 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 attributesrdquo
Types of process validation
Depending on the time when validation is performed relative to the production process validations can be classified as
A) Prospective B) Concurrent
C) Retrospective D) Revalidation
Phases in process validationPhase 1(Pre-validation qualification phase) which covers all activities relating to product research and development formulation pilot batch studies scale-up studies establishing stability conditions and storage and handling of in-process and finished dosage forms equipment qualification installation qualification master production document operational qualification and process capacity
Phase 2 (Process validation phase) It is designed to verify that all established limits of the critical process parameter are valid and satisfactory Products can be produced even under set parameters of the worst case conditions Phase 3 (Validation maintenance Phase) it requires frequent review of all process related documents including validation of audit reports to assure that there have been no changes deviations failures and modifications to the production process and that all standard operating procedures (SOPs) including change control procedures
Purpose of Validation-Minimize reliance on end product testing-To build sterility into a product-Increase SAL to all units-To provide greater assurance and support of the end product sterility testing
Lack of Sterility Assurance
Number of
Recalls
Fiscal Year
-Lack of Sterility Assurance is the reason for drug recalls in last 5 years
-Nearly all drugs recalled due to Lack of Sterility Assurance in last 20 years were produced via aseptic processing
Aseptic Processing
Essential Elements
1
2
3
4
5
6
7
Manufacturing Environment Facility - Room classification
Class (05μ partsft3) ISO Designation WHO Grade
100 5 A
1000 6 B
10000 7 C
100000 8 D
Facility1
Four grades of clean areasGrade D (equivalent to Class 100000 ISO 8)
bull Clean area for carrying out less critical stages in manufacture of aseptically prepared products eg handling of components after washing
Grade C (equivalent to Class 10000 ISO 7)bull Clean area for carrying out less critical stages in
manufacture of aseptically prepared products eg preparation of solutions to be filtered
Grade B (equivalent to Class 1000 ISO 6)
bull Background environment for Grade A zone eg clean room in which laminar flow workstation is housed Class 10000 cleanroom
ndash Local zone for high risk operations eg product filling stopper bowls open vials handling sterile materials aseptic connections transfer of partially stoppered containers to be lyophilized
ndash Conditions usually provided by laminar air flow workstation
bullGrade A (equivalent to Class 100 ISO 5)
Class 100 cleanroom
Airflow patternsThere are two types of aseptic processing isolators =
1Closed isolators employ connections with auxiliary equipment for material transfer Turbulent flow can be acceptable within closed isolators -Gloves half suits seals gaskets and transfer systems should be covered by PM program Physical integrity test routinely performed
2 Open isolators have openings to the environment from the surrounding room via overpressure
Use pressure differential to insure separation of critical area from external environment (175-50 Pa 007-020 water gauge)
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Contents
IntroductionAseptic sterile- ldquo A state of control attained by using an aseptic work area and performing activities in a manner that precludes microbiological contamination of the exposed sterile productrdquo
Process Validation- ldquoProcess validation is establishing documented evidence which demonstrate that the manufacturing process will consistently produce a product meeting its predeterminespecifications and quality Characteristicsrdquo
Drug product container and closure are subject to sterilization separately and then brought together
Validation- is 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 attributesrdquo
Types of process validation
Depending on the time when validation is performed relative to the production process validations can be classified as
A) Prospective B) Concurrent
C) Retrospective D) Revalidation
Phases in process validationPhase 1(Pre-validation qualification phase) which covers all activities relating to product research and development formulation pilot batch studies scale-up studies establishing stability conditions and storage and handling of in-process and finished dosage forms equipment qualification installation qualification master production document operational qualification and process capacity
Phase 2 (Process validation phase) It is designed to verify that all established limits of the critical process parameter are valid and satisfactory Products can be produced even under set parameters of the worst case conditions Phase 3 (Validation maintenance Phase) it requires frequent review of all process related documents including validation of audit reports to assure that there have been no changes deviations failures and modifications to the production process and that all standard operating procedures (SOPs) including change control procedures
Purpose of Validation-Minimize reliance on end product testing-To build sterility into a product-Increase SAL to all units-To provide greater assurance and support of the end product sterility testing
Lack of Sterility Assurance
Number of
Recalls
Fiscal Year
-Lack of Sterility Assurance is the reason for drug recalls in last 5 years
-Nearly all drugs recalled due to Lack of Sterility Assurance in last 20 years were produced via aseptic processing
Aseptic Processing
Essential Elements
1
2
3
4
5
6
7
Manufacturing Environment Facility - Room classification
Class (05μ partsft3) ISO Designation WHO Grade
100 5 A
1000 6 B
10000 7 C
100000 8 D
Facility1
Four grades of clean areasGrade D (equivalent to Class 100000 ISO 8)
bull Clean area for carrying out less critical stages in manufacture of aseptically prepared products eg handling of components after washing
Grade C (equivalent to Class 10000 ISO 7)bull Clean area for carrying out less critical stages in
manufacture of aseptically prepared products eg preparation of solutions to be filtered
Grade B (equivalent to Class 1000 ISO 6)
bull Background environment for Grade A zone eg clean room in which laminar flow workstation is housed Class 10000 cleanroom
ndash Local zone for high risk operations eg product filling stopper bowls open vials handling sterile materials aseptic connections transfer of partially stoppered containers to be lyophilized
ndash Conditions usually provided by laminar air flow workstation
bullGrade A (equivalent to Class 100 ISO 5)
Class 100 cleanroom
Airflow patternsThere are two types of aseptic processing isolators =
1Closed isolators employ connections with auxiliary equipment for material transfer Turbulent flow can be acceptable within closed isolators -Gloves half suits seals gaskets and transfer systems should be covered by PM program Physical integrity test routinely performed
2 Open isolators have openings to the environment from the surrounding room via overpressure
Use pressure differential to insure separation of critical area from external environment (175-50 Pa 007-020 water gauge)
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
IntroductionAseptic sterile- ldquo A state of control attained by using an aseptic work area and performing activities in a manner that precludes microbiological contamination of the exposed sterile productrdquo
Process Validation- ldquoProcess validation is establishing documented evidence which demonstrate that the manufacturing process will consistently produce a product meeting its predeterminespecifications and quality Characteristicsrdquo
Drug product container and closure are subject to sterilization separately and then brought together
Validation- is 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 attributesrdquo
Types of process validation
Depending on the time when validation is performed relative to the production process validations can be classified as
A) Prospective B) Concurrent
C) Retrospective D) Revalidation
Phases in process validationPhase 1(Pre-validation qualification phase) which covers all activities relating to product research and development formulation pilot batch studies scale-up studies establishing stability conditions and storage and handling of in-process and finished dosage forms equipment qualification installation qualification master production document operational qualification and process capacity
Phase 2 (Process validation phase) It is designed to verify that all established limits of the critical process parameter are valid and satisfactory Products can be produced even under set parameters of the worst case conditions Phase 3 (Validation maintenance Phase) it requires frequent review of all process related documents including validation of audit reports to assure that there have been no changes deviations failures and modifications to the production process and that all standard operating procedures (SOPs) including change control procedures
Purpose of Validation-Minimize reliance on end product testing-To build sterility into a product-Increase SAL to all units-To provide greater assurance and support of the end product sterility testing
Lack of Sterility Assurance
Number of
Recalls
Fiscal Year
-Lack of Sterility Assurance is the reason for drug recalls in last 5 years
-Nearly all drugs recalled due to Lack of Sterility Assurance in last 20 years were produced via aseptic processing
Aseptic Processing
Essential Elements
1
2
3
4
5
6
7
Manufacturing Environment Facility - Room classification
Class (05μ partsft3) ISO Designation WHO Grade
100 5 A
1000 6 B
10000 7 C
100000 8 D
Facility1
Four grades of clean areasGrade D (equivalent to Class 100000 ISO 8)
bull Clean area for carrying out less critical stages in manufacture of aseptically prepared products eg handling of components after washing
Grade C (equivalent to Class 10000 ISO 7)bull Clean area for carrying out less critical stages in
manufacture of aseptically prepared products eg preparation of solutions to be filtered
Grade B (equivalent to Class 1000 ISO 6)
bull Background environment for Grade A zone eg clean room in which laminar flow workstation is housed Class 10000 cleanroom
ndash Local zone for high risk operations eg product filling stopper bowls open vials handling sterile materials aseptic connections transfer of partially stoppered containers to be lyophilized
ndash Conditions usually provided by laminar air flow workstation
bullGrade A (equivalent to Class 100 ISO 5)
Class 100 cleanroom
Airflow patternsThere are two types of aseptic processing isolators =
1Closed isolators employ connections with auxiliary equipment for material transfer Turbulent flow can be acceptable within closed isolators -Gloves half suits seals gaskets and transfer systems should be covered by PM program Physical integrity test routinely performed
2 Open isolators have openings to the environment from the surrounding room via overpressure
Use pressure differential to insure separation of critical area from external environment (175-50 Pa 007-020 water gauge)
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Types of process validation
Depending on the time when validation is performed relative to the production process validations can be classified as
A) Prospective B) Concurrent
C) Retrospective D) Revalidation
Phases in process validationPhase 1(Pre-validation qualification phase) which covers all activities relating to product research and development formulation pilot batch studies scale-up studies establishing stability conditions and storage and handling of in-process and finished dosage forms equipment qualification installation qualification master production document operational qualification and process capacity
Phase 2 (Process validation phase) It is designed to verify that all established limits of the critical process parameter are valid and satisfactory Products can be produced even under set parameters of the worst case conditions Phase 3 (Validation maintenance Phase) it requires frequent review of all process related documents including validation of audit reports to assure that there have been no changes deviations failures and modifications to the production process and that all standard operating procedures (SOPs) including change control procedures
Purpose of Validation-Minimize reliance on end product testing-To build sterility into a product-Increase SAL to all units-To provide greater assurance and support of the end product sterility testing
Lack of Sterility Assurance
Number of
Recalls
Fiscal Year
-Lack of Sterility Assurance is the reason for drug recalls in last 5 years
-Nearly all drugs recalled due to Lack of Sterility Assurance in last 20 years were produced via aseptic processing
Aseptic Processing
Essential Elements
1
2
3
4
5
6
7
Manufacturing Environment Facility - Room classification
Class (05μ partsft3) ISO Designation WHO Grade
100 5 A
1000 6 B
10000 7 C
100000 8 D
Facility1
Four grades of clean areasGrade D (equivalent to Class 100000 ISO 8)
bull Clean area for carrying out less critical stages in manufacture of aseptically prepared products eg handling of components after washing
Grade C (equivalent to Class 10000 ISO 7)bull Clean area for carrying out less critical stages in
manufacture of aseptically prepared products eg preparation of solutions to be filtered
Grade B (equivalent to Class 1000 ISO 6)
bull Background environment for Grade A zone eg clean room in which laminar flow workstation is housed Class 10000 cleanroom
ndash Local zone for high risk operations eg product filling stopper bowls open vials handling sterile materials aseptic connections transfer of partially stoppered containers to be lyophilized
ndash Conditions usually provided by laminar air flow workstation
bullGrade A (equivalent to Class 100 ISO 5)
Class 100 cleanroom
Airflow patternsThere are two types of aseptic processing isolators =
1Closed isolators employ connections with auxiliary equipment for material transfer Turbulent flow can be acceptable within closed isolators -Gloves half suits seals gaskets and transfer systems should be covered by PM program Physical integrity test routinely performed
2 Open isolators have openings to the environment from the surrounding room via overpressure
Use pressure differential to insure separation of critical area from external environment (175-50 Pa 007-020 water gauge)
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Phases in process validationPhase 1(Pre-validation qualification phase) which covers all activities relating to product research and development formulation pilot batch studies scale-up studies establishing stability conditions and storage and handling of in-process and finished dosage forms equipment qualification installation qualification master production document operational qualification and process capacity
Phase 2 (Process validation phase) It is designed to verify that all established limits of the critical process parameter are valid and satisfactory Products can be produced even under set parameters of the worst case conditions Phase 3 (Validation maintenance Phase) it requires frequent review of all process related documents including validation of audit reports to assure that there have been no changes deviations failures and modifications to the production process and that all standard operating procedures (SOPs) including change control procedures
Purpose of Validation-Minimize reliance on end product testing-To build sterility into a product-Increase SAL to all units-To provide greater assurance and support of the end product sterility testing
Lack of Sterility Assurance
Number of
Recalls
Fiscal Year
-Lack of Sterility Assurance is the reason for drug recalls in last 5 years
-Nearly all drugs recalled due to Lack of Sterility Assurance in last 20 years were produced via aseptic processing
Aseptic Processing
Essential Elements
1
2
3
4
5
6
7
Manufacturing Environment Facility - Room classification
Class (05μ partsft3) ISO Designation WHO Grade
100 5 A
1000 6 B
10000 7 C
100000 8 D
Facility1
Four grades of clean areasGrade D (equivalent to Class 100000 ISO 8)
bull Clean area for carrying out less critical stages in manufacture of aseptically prepared products eg handling of components after washing
Grade C (equivalent to Class 10000 ISO 7)bull Clean area for carrying out less critical stages in
manufacture of aseptically prepared products eg preparation of solutions to be filtered
Grade B (equivalent to Class 1000 ISO 6)
bull Background environment for Grade A zone eg clean room in which laminar flow workstation is housed Class 10000 cleanroom
ndash Local zone for high risk operations eg product filling stopper bowls open vials handling sterile materials aseptic connections transfer of partially stoppered containers to be lyophilized
ndash Conditions usually provided by laminar air flow workstation
bullGrade A (equivalent to Class 100 ISO 5)
Class 100 cleanroom
Airflow patternsThere are two types of aseptic processing isolators =
1Closed isolators employ connections with auxiliary equipment for material transfer Turbulent flow can be acceptable within closed isolators -Gloves half suits seals gaskets and transfer systems should be covered by PM program Physical integrity test routinely performed
2 Open isolators have openings to the environment from the surrounding room via overpressure
Use pressure differential to insure separation of critical area from external environment (175-50 Pa 007-020 water gauge)
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Purpose of Validation-Minimize reliance on end product testing-To build sterility into a product-Increase SAL to all units-To provide greater assurance and support of the end product sterility testing
Lack of Sterility Assurance
Number of
Recalls
Fiscal Year
-Lack of Sterility Assurance is the reason for drug recalls in last 5 years
-Nearly all drugs recalled due to Lack of Sterility Assurance in last 20 years were produced via aseptic processing
Aseptic Processing
Essential Elements
1
2
3
4
5
6
7
Manufacturing Environment Facility - Room classification
Class (05μ partsft3) ISO Designation WHO Grade
100 5 A
1000 6 B
10000 7 C
100000 8 D
Facility1
Four grades of clean areasGrade D (equivalent to Class 100000 ISO 8)
bull Clean area for carrying out less critical stages in manufacture of aseptically prepared products eg handling of components after washing
Grade C (equivalent to Class 10000 ISO 7)bull Clean area for carrying out less critical stages in
manufacture of aseptically prepared products eg preparation of solutions to be filtered
Grade B (equivalent to Class 1000 ISO 6)
bull Background environment for Grade A zone eg clean room in which laminar flow workstation is housed Class 10000 cleanroom
ndash Local zone for high risk operations eg product filling stopper bowls open vials handling sterile materials aseptic connections transfer of partially stoppered containers to be lyophilized
ndash Conditions usually provided by laminar air flow workstation
bullGrade A (equivalent to Class 100 ISO 5)
Class 100 cleanroom
Airflow patternsThere are two types of aseptic processing isolators =
1Closed isolators employ connections with auxiliary equipment for material transfer Turbulent flow can be acceptable within closed isolators -Gloves half suits seals gaskets and transfer systems should be covered by PM program Physical integrity test routinely performed
2 Open isolators have openings to the environment from the surrounding room via overpressure
Use pressure differential to insure separation of critical area from external environment (175-50 Pa 007-020 water gauge)
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Aseptic Processing
Essential Elements
1
2
3
4
5
6
7
Manufacturing Environment Facility - Room classification
Class (05μ partsft3) ISO Designation WHO Grade
100 5 A
1000 6 B
10000 7 C
100000 8 D
Facility1
Four grades of clean areasGrade D (equivalent to Class 100000 ISO 8)
bull Clean area for carrying out less critical stages in manufacture of aseptically prepared products eg handling of components after washing
Grade C (equivalent to Class 10000 ISO 7)bull Clean area for carrying out less critical stages in
manufacture of aseptically prepared products eg preparation of solutions to be filtered
Grade B (equivalent to Class 1000 ISO 6)
bull Background environment for Grade A zone eg clean room in which laminar flow workstation is housed Class 10000 cleanroom
ndash Local zone for high risk operations eg product filling stopper bowls open vials handling sterile materials aseptic connections transfer of partially stoppered containers to be lyophilized
ndash Conditions usually provided by laminar air flow workstation
bullGrade A (equivalent to Class 100 ISO 5)
Class 100 cleanroom
Airflow patternsThere are two types of aseptic processing isolators =
1Closed isolators employ connections with auxiliary equipment for material transfer Turbulent flow can be acceptable within closed isolators -Gloves half suits seals gaskets and transfer systems should be covered by PM program Physical integrity test routinely performed
2 Open isolators have openings to the environment from the surrounding room via overpressure
Use pressure differential to insure separation of critical area from external environment (175-50 Pa 007-020 water gauge)
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Manufacturing Environment Facility - Room classification
Class (05μ partsft3) ISO Designation WHO Grade
100 5 A
1000 6 B
10000 7 C
100000 8 D
Facility1
Four grades of clean areasGrade D (equivalent to Class 100000 ISO 8)
bull Clean area for carrying out less critical stages in manufacture of aseptically prepared products eg handling of components after washing
Grade C (equivalent to Class 10000 ISO 7)bull Clean area for carrying out less critical stages in
manufacture of aseptically prepared products eg preparation of solutions to be filtered
Grade B (equivalent to Class 1000 ISO 6)
bull Background environment for Grade A zone eg clean room in which laminar flow workstation is housed Class 10000 cleanroom
ndash Local zone for high risk operations eg product filling stopper bowls open vials handling sterile materials aseptic connections transfer of partially stoppered containers to be lyophilized
ndash Conditions usually provided by laminar air flow workstation
bullGrade A (equivalent to Class 100 ISO 5)
Class 100 cleanroom
Airflow patternsThere are two types of aseptic processing isolators =
1Closed isolators employ connections with auxiliary equipment for material transfer Turbulent flow can be acceptable within closed isolators -Gloves half suits seals gaskets and transfer systems should be covered by PM program Physical integrity test routinely performed
2 Open isolators have openings to the environment from the surrounding room via overpressure
Use pressure differential to insure separation of critical area from external environment (175-50 Pa 007-020 water gauge)
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Four grades of clean areasGrade D (equivalent to Class 100000 ISO 8)
bull Clean area for carrying out less critical stages in manufacture of aseptically prepared products eg handling of components after washing
Grade C (equivalent to Class 10000 ISO 7)bull Clean area for carrying out less critical stages in
manufacture of aseptically prepared products eg preparation of solutions to be filtered
Grade B (equivalent to Class 1000 ISO 6)
bull Background environment for Grade A zone eg clean room in which laminar flow workstation is housed Class 10000 cleanroom
ndash Local zone for high risk operations eg product filling stopper bowls open vials handling sterile materials aseptic connections transfer of partially stoppered containers to be lyophilized
ndash Conditions usually provided by laminar air flow workstation
bullGrade A (equivalent to Class 100 ISO 5)
Class 100 cleanroom
Airflow patternsThere are two types of aseptic processing isolators =
1Closed isolators employ connections with auxiliary equipment for material transfer Turbulent flow can be acceptable within closed isolators -Gloves half suits seals gaskets and transfer systems should be covered by PM program Physical integrity test routinely performed
2 Open isolators have openings to the environment from the surrounding room via overpressure
Use pressure differential to insure separation of critical area from external environment (175-50 Pa 007-020 water gauge)
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
ndash Local zone for high risk operations eg product filling stopper bowls open vials handling sterile materials aseptic connections transfer of partially stoppered containers to be lyophilized
ndash Conditions usually provided by laminar air flow workstation
bullGrade A (equivalent to Class 100 ISO 5)
Class 100 cleanroom
Airflow patternsThere are two types of aseptic processing isolators =
1Closed isolators employ connections with auxiliary equipment for material transfer Turbulent flow can be acceptable within closed isolators -Gloves half suits seals gaskets and transfer systems should be covered by PM program Physical integrity test routinely performed
2 Open isolators have openings to the environment from the surrounding room via overpressure
Use pressure differential to insure separation of critical area from external environment (175-50 Pa 007-020 water gauge)
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Airflow patternsThere are two types of aseptic processing isolators =
1Closed isolators employ connections with auxiliary equipment for material transfer Turbulent flow can be acceptable within closed isolators -Gloves half suits seals gaskets and transfer systems should be covered by PM program Physical integrity test routinely performed
2 Open isolators have openings to the environment from the surrounding room via overpressure
Use pressure differential to insure separation of critical area from external environment (175-50 Pa 007-020 water gauge)
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
2 Open isolators have openings to the environment from the surrounding room via overpressure
Use pressure differential to insure separation of critical area from external environment (175-50 Pa 007-020 water gauge)
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Pressure differentials Used to maintain airflow in the direction of higher
cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained
between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
HEPAULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors walls and ceilings constructed of smooth hard
surfaces that can be easily cleaned Limited equipment fixtures and personnel Layout of equipment to optimize comfort and movement
of operators
General Cleanroom Design
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Material NOT permitted in a Cleanroom
Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into non-
cardboard containers Wood (ie wooden pallets)
1 Vacuum all accessible surfaces
2 Wipe surfaces with a cleaning solution3 Mop floors using a lint free polyester mops attached to stainless steel handles
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
2 Design
Differential Pressure - 10-15 Pascals
Temperature ndash 18-20degC
Relative Humidity ndash 30-60 RH
Material Flow -Entry through air lock system or sterilizing ovens or filters
-Contact plate testing should be done for assurance
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Equipment2
Contact sterile materials only with sterile instruments-Sterile instruments should be held under Class 100 conditions
between uses and placed in sterile containers-Operators should not contact sterile products containers closures
or critical surfaces with any part of their gown or gloves-Equipment includes tanks centrifuges and dryers is intended to be sterilize before use -The validation program should show the effectiveness of disinfecting program-It also prove that steam is sterilizing all surfaces-Heat distribution study should be done to determine the cold spots where condensate could accumulate
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Process3
Process flow of parenteral dosage form
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Process3
- In aseptic processing each component is individually sterilised or several components are combined with the resulting mixture sterilized
-Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers biological products
-Media fill program should include worst case activities
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered preparation should be prepared in a Grade A environment with Grade B background (eg ointments creams suspensions and emulsions) Prepared solutions filtered through a sterile 022μm (or less) membrane filter into a previously sterilized container
filters remove bacteria and moulds do not remove all viruses or mycoplasmas
filtration should be carried out under positive pressurePreparation and Filtration of Solutions (2) consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Fitlers should not shed particles asbestos containing filters should not be used Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels pressure release outlets should have hydrophobic microbial retentive air filters Preparation and Filtration of Solutions (3) Time limits should be established for each phase of processing eg
maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containerscomponents total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment)
Preparation and Filtration of Solutions (4) Filling of solution may be followed by lyophilization (freeze drying)
stoppers partially seated product transferred to lyophilizer (Grade AB conditions) Release of airnitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established
and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achievedPrefiltation Bioburden (2) No defined ldquomaximumrdquo limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in ldquoin-processrdquo controls
particularly when product supports microbial growth andor manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxinspyrogens
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Filter integrity Filters of 022μm or less should be used for filtration of liquids and gasses (if applicable)
filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration
bubble point pressure hold forward flow
methods are defined by filter manufacturers and limits determined during filter validationFilter Validaton Filter must be validated to demonstrate ability to remove bacteria
most common method is to show that filter can retain a microbiological challenge of 107 CFU of Brevundimonas diminuta per cm2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter
Filter validation (2) if the product is bactericidal product should be passed through the filter first followed by modified product containing the microbial challenge (after
removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions eg maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
Equipmentcontainer preparation and sterilization All equipment (including lyophilizers) and product containersclosures should be sterilized using validated cycles
same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters housings and tubing
Equipmentcontainer preparation and sterilization (2) CIPSIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP heat tunnels often used for sterilizationdepyrogenation of glass vialsbottles
usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units)
worst case locations tunnel supplied with HEPA filtered air
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Equipmentcontainer preparation and sterilization (2) equipment should be designed to be easily assembled and disassembled cleaned sanitised andor sterilized
equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified)Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill)
Process simulation includes formulation (compounding) filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats eg lyophilized products ointments sterile bulks eye drops filled into semi-transparentopaque containers
biological products
Process Validation (2) Media fill program should include worst case activities
Factors associated with longest permitted run (eg operator fatigue) Representative number type and complexity of normal interventions non-routine interventions and events (eg maintenance stoppages etc) Lyophilisation Aseptic equipment assembly
Process Validation (3) Worst case activities (cont)
No of personnel and their activities shift changes breaks gown changes Representative number of aseptic additions (eg charging containers closures sterile ingredients) or transfers Aseptic equipment connectionsdisconnections Aseptic sample collections Line speed and configuration
Process Validation (4) Worst case activities (cont)
Weight checks Container closure systems Specific provisions in processing instructions
Written batch record documenting conditions and activities Should not be used to justify risky practices
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
- Preparation and Filtration of Solutions
- Prefiltration Bio-burden (natural microbial load)
- Filter integrity
- Filter Validaton
- Equipmentcontainer preparation and sterilization
- Process Validation
Aseptic Processing
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
- Duration-Depends on type of operation-For conventional operations should include the total filling time
- Size-5000 - 10000 generally acceptable or batch size if lt5000-For manually intensive processes larger numbers should be filled-Lower numbers can be filled for isolators
- Line Speed-Speed depends on type of process
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
1st ndash eliminate the source of contamination 2nd - Reduce the Risk of contamination through
Sterile barriers Aseptic techniqueEnvironmental monitoring
Personnel4
3rd Gowning Gloves
-Frequent disinfection of gloves shoud done during operations
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
-Gowning process must be validated
-Done by Surface Monitoring or microbiological monitoring (Touch- swab or Contact plates- RODAC Plates) Glove fingertips into Petri dish containing agar media
-Change gloves and masks at every working session
-No watches jewellery and cosmetics-Eye checks if involved in visual inspection
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Personnel Hygiene
Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder hair sprays
perfumes and aftershave Clothing should be clean nonfrayed and nonlinting Avoid smoking
Behavior-Minimize movement Work slowly and purposefully
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Control amp Verification5
1 Environmental Monitoring
A] Surface monitoring Product contact surfaces floors walls and equipment should be tested on a regular basis
B] Active air monitoring testing of the number of organisms per volume of air sampled
C] Passive air monitoring settling plates can be used as qualitative or semi-quantitative air monitors
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Control amp Verification5
1 Environmental Monitoring -Physical-Particulate matter =
-Differential pressures
-Air changes airflow patterns
-Clean up timerecovery
-Temperature and relative humidity
-Airflow velocityThe goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends and historical data
Particulate Air MonitoringUse of remote systems recommended in laminar flow areas
ldquoAt restrdquo - production equipment installed and operating
ldquoIn operationrdquo - Installed equipment functioning in defined operating mode and specified number of personnel present
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
2 Personnel Monitoring
-Clothing of appropriate quality Grade D
hair beard moustache covered protective clothing and shoes
Grade C hair beard moustache covered single or 2-piece suit (covering wrists high neck)
shoesovershoes no fibresparticles to be shed
Grade A and B headgear beard and moustache covered masks gloves not shedding fibres and retain particles shed by operators
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Environmental Monitoring - Physical Particulate matter
Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite within airflow and during fillingclosing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (eg powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
Environmental Monitoring - Physical Differential pressures
Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded Alarms should sound if pressures deviate Any deviations should be investigated and effect on environmental quality determined
Environmental Monitoring - Physical Air ChangesAirflow patterns
Air flow over critical areas should be uni-directional (laminar flow) at a velocity sufficient to sweep particles away from fillingclosing area for B C and D rooms at least 20 changes per hour are ususally required
Clean up timerecovery Particulate levels for the Grade A ldquoat restrdquo state should be achieved after a short ldquoclean-uprdquo period of 20 minutes after completion of operations (guidance
value) Particle counts for Grade A ldquoin operationrdquo state should be maintained whenever product or open container is exposed
Environmental Monitoring - Physical Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18degC) Airflow velocity
Laminar airflow workstation air speed of approx 045ms plusmn 20 at working position (guidance value)Personnel Minimum number of personnel in clean areas
especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff
initial and regular manufacturing hygiene microbiology should be formally validated and authorized to enter aseptic area
Special cases supervision in case of outside staff decontamination procedures (eg staff who worked with animal tissue materials)
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Personnel (2) High standards of hygiene and cleanliness
should not enter clean rooms if ill or with open wounds Periodic health checks No shedding of particles movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas should be clad in factory clothing Changing and washing procedure No watches jewellery and cosmetics Eye checks if involved in visual inspectionPersonnel (4) Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments ndash separate laundry facility
No damage and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
3 Aseptic Filling Simulations (Media Fills) Verification of medium sterility Aseptic filling operation Challenge unit incubation Evaluation of resultMedia used- Soybean casein digest media Fluid thioglycolate mediaMedia fill frequency- 6 months interval If change in aseptic processAcceptance criteria- Historically 03(as per WHO) 01 as per Parentral Drug Association (PDA)
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Acceptance Criteria1048708 Ideally the contamination rate should be zero However currently theaccepted contamination rate should be less than 01 with a 95 confidence level according to the Annex I to the EUPICS Guide toGMP
Incubation Temperature1048708 It is generally accepted to incubate at 20-25degC for aminimum of 14 days without having collected data tosupport this incubation schedule1048708 It is similarly acceptable for firms who prefer a twotemperature incubation schedule to incubate at 20-25degCfor a minimum of 7 days followed immediately byincubation at a higher temperature range not to exceed35degC for a total minimum incubation time of 14 days
Process simulation studies (media fills) are simulating thewhole process in order to evaluate the sterility confidenceof the processProcess simulation studies include formulation(compounding) filtration and filling with suitable mediaSimulations are made to ensure that the regular process forcommercial batches repeatedly and reliably produces thefinished product of the required quality However eachprocess simulation trial is unique and so it is not possible toextrapolate these results directly to actual productioncontamination rates
Where filling takes place over extended periods ielonger than 24 hours the process simulation test shouldextend over the whole of the standard filling period Inorder to prevent excessively high numbers of units beingfilled it is usually acceptable to just run the machine for areasonable time if the validity of the simulation is notdiminished by this procedure1048708 The fill volume of the containers should be sufficient toenable contact of all the container-closure seal surfaceswhen the container is inverted and also sufficient to allowthe detection of microbial growth
Process simulation studies (media fills)
Media fills should be observed by the QC unit Video recording should be considered
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Finish Product amp Testing6
Sterility TestingParticulate TestingContainer Closure Integrity TestingOther Final ProductRelease TestingStability Testing
Finish Product Testing after Sterilization1048708Uniformity of filled volume1048708 Perform testing on filled containers1048708Sterility1048708 10 samples from each of the beginning and end of the filling runSamples must represent all filling nozzles1048708 Visual Evaluation1048708 Appearance Color of solution1048708 Other Testing1048708 Assay pH Density Pyrogen or Endotoxin etc
Thesterilitytestcanprovideusefulinformationonthevalidationstatusofasepticprocess
ThesterilitytestappliedtothefinishedproductshouldonlyberegardedasthelastinaseriesofcontrolmeasuresbywhichsterilityisassuredThetestshouldbevalidatedfortheproduct(s)concerned
Endotoxin a pyrogenic (fever inducing) substance (eg lipopolysaccharide) present in the bacterial cell wall Endotoxin reactions range from fever to
death
21 CFR 211167ldquo For each batch of drug product purporting to be sterile andor pyrogen-free there shall be appropriate laboratory testing to determine conformance to such requirementsrdquo
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Documentation7Media Fill RecordsProduction Batch RecordsEM Trend DataRelease Testing Batch RecordsInvestigation
Response to ExcursionsCorrective Actions
Validation Report1048708 Validation Team must prepare the report1048708 Report must be reviewed and approved by QA1048708 Written Notification or either successful completion or failure of theprocess validation must be issued to top management1048708 In case of failure an investigation must be completed and documentedprior to repeat the validation study
Sterility Testing Investigations
In the investigation stemming from a negative sterility test consideration should be given to Speciation of the organism Record of laboratory results and deviations Environmental monitoring of production environment Monitoring personnel Product Presterilization bioburden Production record review Manufacturing history
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Process Validation of Sterile Liquid Products
Validation Team Production QC QA EngineerPlanner
bullTo prepare the validation protocolbullVerify the calibration and maintenance status of equipmentbullPerform qualification for equipments and systembull Verify change controlbull Schedule the validation activitiesbull Training production operatorsbull Conduct validation studybull Monitor the critical steps in manufacturing processbull Assure that the approved testing standard is being usedbull Evaluate all test resultsbull Prepare the validation report
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
Pre-validation Requirements
bullPreventive Maintenance for Facilities and Utilitiesbull Calibration of Equipmentbull Cleaning Validationbull Equipment amp System Qualificationbull Raw MaterialsComponentsTest Methodsbull Process Justificationbull Change Controlbull Training operators
All must be proven suitable and reliable for the manufacturing process before the process can be validated
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
New Product lt==gt Trial Batch Development Batch Transferred Product lt==gt Products produced at the sending site Revalidation Product lt==gt The original product before revalidationType of Process Validation1 Prospectiveo Conducted prior to market the product2 Concurrento Based on information generated during actualimplementation of the process (each batch will bereleased separately)3 Retrospective (Not recommended for sterile product)O Based on accumulated historical production testing andcontrol dataO Generally requires data from 10-30 batchesO Use data only from batches made by the same process
Terminal Sterilization Operation whereby the product is sterilized separately byautoclave after filled and packaged using sterilized containersand closures in critical processing zonesAseptic OperationOperation whereby the product is sterilized separately by filteringthrough 02 μ or less filter then filled and packaged usingsterilized containers and closures in critical processing zones
Process Justification1048708 To identify critical process steps amp process parameter of Mixingprocess1048708 To determine the suitable Hold time Period1048708 To confirm the analytical tests that will have to be performed1048708 To define the optimal parameters throughout the overall ampoulefilling process to consistently produce the finished products(filledampoules) which meet the established specifications1048708 To assure that the product is sterile after sterilization process
Validation ProtocolA document stating how validation will be conductedincluding test parameters product characteristics productionequipment to be used and decision points on whatconstitutes acceptable test results
Validation Protocol should contain 1048708 Title Page ReviewApproval Page1048708 Purpose and Overview1048708 Equipment List1048708 Ingredients and Component List1048708 Qualification List of Equipment and System1048708 Process Flow Diagram and Description1048708 Equipment Critical Process Parameter1048708 Process Validation Sampling PlanTesting Requirements1048708 Acceptance Criteria1048708 Stability Requirements1048708 Process for evaluation of any deviations occurring during validation1048708 Conclusion
Equipment Critical Process Parameter1048708 Mixing Speed1048708 Mixing Time1048708 Gas flushing time1048708 Type and size of filter1048708 Filtering Time and Pressure used1048708 Filling Speed1048708 Temperature and Duration for Terminal SterilizationCritical Manufacturing Step1048708 Dissolving Step1048708 pH adjustment step1048708 Final mixing step1048708 Filtering Step1048708 Filling Step1048708 Terminal Sterilization Step1048708 Leak Test Step
Changes and Revalidation1048708 Change of any of the following may need revalidation1048708 Formula Composition1048708 Raw Material Source1048708 Manufacturing Process1048708 Manufacturing Location1048708 Equipments1048708 Batch Size1048708 Testing Specification
Changes1048708 Minor It seems to have no impact on formulation1048708 It is not necessary to validate1048708 Intermediate It could have significant impact on formulation1048708 Depend on case-by-case (A minimum of 1 trial)1048708 Major It is likely to have significant impact on formulation1048708 Revalidation is required (A minimum of 3 trials)Validation
Minor Change1048708 Qualitative inactive excipient change deemed minor bychange control review1048708 Process change deemed minor by change controlreview1048708 Manufacturing location change with in same buildingsame equipment personnel procedure and utilities areused1048708 Equipment change but same design configuration
Intermediate Change1048708 Process changes such as mixing times or operating speedsfor solutions1048708 Change in release specification to a tighter limit causedoriginal validation results to be out of specification1048708 Extension of the qualified in process hold time for intermediateor finished product prior to packaging1048708 Equipment change deemed intermediate by change controlreview
Major Changes1048708 Quantitative or qualitative formulation change deemed major bychange control review1048708 Inactive excipient or active ingredient source change deemedmajor by change control review1048708 Transfer product from on site to another1048708 Significant change in process1048708 Equipment change to a different design configuration or operatingprinciple
