Aseptic processing
- 1. ASEPTIC PROCESSING TECHNOLOGY From : Riya R Yadav Roll no : MQA 07 NMIMS SHIRPUR
- 2. ASEPTIC PROCESSING- OVERVIEW Certain pharmaceutical products must be sterile – injections, ophthalmic preparations, irrigations solutions, haemodialysis solutions Two categories of sterile products – those that can be sterilized in final container (terminally sterilized) – those that cannot be terminally sterilized and must be aseptically prepared
- 3. OVERVIEW ASEPTIC PROCESSING Objective is to maintain the sterility of a product, assembled from sterile components • Operating conditions so as to prevent microbial contamination
- 4. OVERVIEW To review specific issues relating to the manufacture of aseptically prepared products: – Manufacturing environment Clean areas Personnel – Preparation and filtration of solutions – Pre-filtration bioburden – Filter integrity/validation – Equipment/container preparation and sterilization – Filling Process – Validation of aseptic processes – Specific issues relating to Isolators, BFS and Bulk
- 5. MANUFACTURING ENVIRONMENT Classification of Clean Areas -Comparison of classifications.
- 6. MANUFACTURING ENVIRONMENT CLASSIFICATION OF CLEAN AREAS Classified in terms of airborne particles “At rest” - production equipment installed and operating “In operation” - Installed equipment functioning in defined operating mode and specified number of personnel present
- 7. MANUFACTURING ENVIRONMENT FOUR GRADES OF CLEAN AREAS: Grade D (equivalent to Class 100,000, ISO 8): – 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 10,000, ISO 7): – 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 100, ISO 5): – Background environment for Grade A zone, eg . cleanroom in which laminar flow workstation is housed.
- 8. MANUFACTURING ENVIRONMENT Grade A (equivalent to Class 100 (US Federal Standard 209E), ISO 5 (ISO 14644-1): – 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. – Conditions usually provided by laminar air flow workstation. -Each grade of cleanroom has specifications for viable and non-viable particles – Non-viable particles are defined by the air classification
- 9. MANUFACTURING ENVIRONMENT Limits for viable particle ( microbiological contamination) Table 3 – These are average values – Individual settle plates may be exposed for less than 4 hours • Values are for guidance only - not intended to represent specifications • Levels (limits) of detection of microbiological contamination should be established for alert and action purposes and for monitoring trends of air quality in the facility
- 10. MANUFACTURING ENVIRONMENT Environmental Monitoring Physical – Particulate matter – Differential pressures – Air changes, airflow patterns – Clean up time/recovery – Temperature and relative humidity – Airflow velocity
- 11. MANUFACTURING ENVIRONMENT 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 filling/closing operations – Preferably a remote probe that monitors continuously – Difficulties when process itself generates particles (e.g. powder filling) – Appropriate alert and action limits should be set and corrective actions defined if limits exceeded
- 12. MANUFACTURING ENVIRONMENT 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
- 13. MANUFACTURING ENVIRONMENT Environmental Monitoring Physical • Air Changes/Airflow patterns – Air flow over critical areas should be uni -directional (laminar flow) at a velocity sufficient to sweep particles away from filling/closing area – for B, C and D rooms at least 20 changes per hour are ususally required Clean up time/recovery - Particulate levels for the Grade A “at rest” state should be achieved after a short “clean-up” period of 20 minutes after completion of operations (guidance value) – Particle counts for Grade A “in operation” state should be maintained whenever product or open container is exposed
- 14. MANUFACTURING ENVIRONMENT Environmental Monitoring Physical • Temperature and Relative Humidity – Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18°C) • Airflow velocity – Laminar airflow workstation air speed of approx. 0.45m/s ± 20% at working position (guidance value)
- 15. MANUFACTURING ENVIRONMENT 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 (e.g. staff who worked with animal tissue materials)
- 16. MANUFACTURING ENVIRONMENT 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 inspection
- 17. MANUFACTURING ENVIRONMENT PERSONNEL (3) • 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), shoes/overshoes • no fibres/particles to be shed – Grade A and B • headgear, beard and moustache covered, masks, gloves • not shedding fibres, and retain particles shed by operators
- 18. MANUFACTURING ENVIRONMENT PERSONNEL (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 – separate laundry facility – No damage, and according to validated procedures (washing and sterilization) • Regular microbiological monitoring of operators
- 19. ASEPTIC PROCESSING In aseptic processing, each component is individually sterilised, or several components are combined with the resulting mixture sterilized – Most common is preparation of a solution which is filtered through a sterilizing filter then filled into sterile containers (eg . active and excipients dissolved in Water for Injection) – May involve aseptic compounding of previously sterilized components which is filled into sterile containers – May involve filling of previously sterilized powder • sterilized by dry heat/irradiation • produced from a sterile filtered solution which is then aseptically crystallized and precipitated – requires more handling and manipulation with higher potential for contamination during processing
- 20. 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 (e.g. ointments, creams, suspensions and emulsions) • Prepared solutions filtered through a sterile 0.22μm (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 pressure
- 21. • Filling of solution may be followed by lyophilization (freeze drying) – stoppers partially seated, product transferred to lyophilizer (Grade A/B conditions) – Release of air/nitrogen into lyophilizer chamber at completion of process should be through sterilizing filter
- 22. PREFILTRATION BIOBURDEN (NATURAL MICROBIAL LOAD) 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 (e.g. ointments, creams, suspensions and emulsions) • Prepared solutions filtered through a sterile 0.22μm (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 pressure
- 23. FILTER INTEGRITY • Filters of 0.22μm 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 validation.
- 24. FILTER 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
- 25. EQUIPMENT/CONTAINER PREPARATION AND STERILIZATION All equipment (including lyophilizers) and product containers/closures 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
- 26. 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 e.g. lyophilized products, ointments, sterile bulks, eye drops filled into semi- transparent/opaque containers, biological products
- 27. ADDITIONAL ISSUES SPECIFIC TO ISOLATOR AND BFS TECHNOLOGIES • Isolators – Decontamination process requires a 4-6 log reduction of appropriate Biological Indicator (BI) – Minimum 6 log reduction of BI if surface is to be free of viable organisms – Significant focus on glove integrity - daily checks, second pair of gloves inside isolator glove – Traditional aseptic vigilance should be maintained
- 28. BLOW-FILL-SEAL (BFS) – Located in a Grade D environment – Critial zone should meet Grade A (microbiological) requirements (particle count requirements may be difficult to meet in operation) – Operators meet Grade C garment requirements – Validation of extrusion process should demonstrate destruction of endotoxin and spore challenges in the polymeric material – Final inspection should be capable of detecting leakers
- 29. ISSUES RELATING TO ASEPTIC BULK PROCESSING • Applies to products which can not be filtered at point of fill and require aseptic processing throughout entire manufacturing process. • Entire aseptic process should be subject to process simulation studies under worst case conditions (maximum duration of "open" operations, maximum no of operators) • Process simulations should incorporate storage and transport of bulk. • Multiple uses of the same bulk with storage in between should also be included in process simulations • Assurance of bulk vessel integrity for specified holding times.
- 30. BULK PROCESSING Process simulation for formulation stage should be performed at least twice per year. – Cellular therapies, cell derived products etc • products released before results of sterility tests known (also TPNs, radioactive preps, cytotoxics) • should be manufactured in a closed system • Additional testing – sterility testing of intermediates – microscopic examination (e.g. gram stain) – endotoxin testing