Aseptic Processing: Mrs Robyn Isaacson

Aseptic Processing
Mrs Robyn Isaacson
Manufacture of sterile medicines Advanced workshop for SFDA GMP inspectors - Nanjing, November 2009
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

Aseptic processing Objective is to maintain the sterility of a product, assembled from sterile components Operating conditions so as to prevent microbial contamination

Objective
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
Manufacturing Environment
Classification of Clean Areas
Comparison of classifications
WHO GMP Grade A Grade B Grade C Grade D
US 209E M 3.5 M 3.5 M 5.5 M 6.5
US Customary Class 100 Class 100 Class 10 000 Class 100 000
ISO/TC (209) ISO 14644 ISO 5 ISO 5 ISO 7 ISO 8
EEC GMP Grade A Grade B Grade C Grade D
Table 1
5
Classification of Clean Areas
Classified in terms of airborne particles (Table 2)
Grade At rest In operation maximum permitted number of particles/m3 0.5 - 5.0 m A B C D 3 500 3 500 350 000 3 500 000 > 5 m 0 0 2 000 20 000 0.5 - 5.0 m 3 500 350 000 3 500 000 not defined >5 0 2 000 20 000 not defined
At rest - production equipment installed and operating In operation - Installed equipment functioning in defined operating mode and specified number of personnel present
6
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.
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 (See Table 2)
Limits for viable particles (microbiological contamination)
Grade Air sample (CFU/m3) Settle plates (90mm Contact plates diameter) (55mm (CFU/4hours) diameter) (CFU/plate) Glove print (5 fingers) (CFU/glove)
A B C D
<3 10 100 200
<3 5 50 100
<3 5 25 50
<3 5 -
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
9
Environmental Monitoring Physical
Particulate matter
Differential pressures Air changes, airflow patterns Clean up time/recovery Temperature and relative humidity Airflow velocity
10
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
11
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
12
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
13
Temperature and Relative Humidity
Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18C)
Airflow velocity
Laminar airflow workstation air speed of approx 0.45m/s 20% at working position (guidance value)
14
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)
15
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
16
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
17
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
18
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 (e.g 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
19
Aseptic Processing
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 0.22m (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

20
Aseptic Processing
Preparation 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
21
Aseptic Processing
Time limits should be established for each phase of processing, e.g.
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 containers/components 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)
22
Aseptic Processing
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
23
Aseptic Processing
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 achieved
24
Aseptic Processing
Prefiltation Bioburden (2)
No defined maximum limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in inprocess controls
particularly when product supports microbial growth and/or manufacturing process involves use of culture media
Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxins/pyrogens
25
Aseptic Processing
Filter integrity
Filters of 0.22m 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
26
Aseptic Processing
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
27
Aseptic Processing
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 e.g. maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation
28
Aseptic Processing
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
29
Aseptic Processing
Equipment/container preparation and sterilization (2)
CIP/SIP processes
particular attention to deadlegs - different orientation requirements for CIP and SIP
heat tunnels often used for sterilization/depyrogenation of glass vials/bottles

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

30
Aseptic Processing
Equipment/container preparation and sterilization (2)
equipment should be designed to be easily assembled and disassembled, cleaned, sanitised and/or 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)
31
Aseptic Processing
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
32
Aseptic Processing
Process Validation (2)
Media fill program should include worst case activities
Factors associated with longest permitted run (e.g. operator fatigue) Representative number, type, and complexity of normal interventions, non-routine interventions and events (e.g. maintenance, stoppages, etc) Lyophilisation Aseptic equipment assembly
33
Aseptic Processing
Worst case activities (cont)
No of personnel and their activities, shift changes, breaks, gown changes Representative number of aseptic additions (e.g. charging containers, closures, sterile ingredients) or transfers Aseptic equipment connections/disconnections Aseptic sample collections Line speed and configuration
34
Aseptic Processing
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
35
Aseptic Processing
Duration
Depends on type of operation BFS, Isolator processes - sufficient time to include manipulations and interventions For conventional operations should include the total filling time
Size
5000 - 10000 generally acceptable or batch size if <5000 For manually intensive processes larger numbers should be filled Lower numbers can be filled for isolators
36
Aseptic Processing
Frequency and Number
Three initial, consecutive per shift
Subsequently semi-annual per shift and process All personnel should participate at least annually, consistent with routine duties Changes should be assessed and revalidation carried out as required
Line Speed
Speed depends on type of process
37
Aseptic Processing
Environmental conditions
Representative of actual production conditions (no. of personnel, activity levels etc) - no special precautions (not including adjustment of HVAC) if nitrogen used for overlaying/purging need to substitute with air
Media
Anaerobic media should be considered under certain circumstances Should be tested for growth promoting properties (including factory isolates)
38
Aseptic Processing
Incubation, Examination
In the range 20-35C.
If two temperatures are used, lower temperature first Inspection by qualified personnel.
All integral units should be incubated. Should be justification for any units not incubated.
Units removed (and not incubated) should be consistent with routine practices (although incubation would give information regarding risk of intervention) Batch reconciliation
39
Aseptic Processing
Interpretation of Results
When filling fewer than 5000 units:
no contaminated units should be detected One (1) contaminated unit is considered cause for revalidation, following an investigation
When filling from 5000-10000 units

One (1) contaminated unit should result in an investigation, including consideration of a repeat media fill Two (2) contaminated units are considered cause for revalidation, following investigation
When filling more than 10000 units

One (1) contaminated unit should result in an investigation Two (2) contaminated units are considered cause for revalidation, following investigation
40
Aseptic Processing
Interpretation of Results
Media fills should be observed by QC and contaminated units reconcilable with time and activity being simulated (Video may help) Ideally - no contamination. Any contamination should be investigated. Any organisms isolated should be identified to species level (genotypic identification) Invalidation of a media fill run should be rare
41
Aseptic Processing
Batch Record Review
Process and environmental control activities should be included in batch records and reviewed as part of batch release

In-process and laboratory control results Environmental and personnel monitoring data Output from support systems(HEPA/HVAC, WFI, steam generator) Equipment function (batch alarm reports, filter integrity) Interventions, Deviations, Stoppages - duration and associated time Written instructions regarding need for line clearances Disruptions to power supply
42
Aseptic Processing
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
43
Aseptic Processing
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
44
Aseptic Processing
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.
45
Aseptic Processing
Bulk Processing (2)
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
46
Useful Publications
PIC/S Recommendation on the Validation of Aseptic Processes FDA Guidance for Industry- Sterile Drug Products Produced by Aseptic Processing - Current Good Manufacturing Process ISO 13408 Aseptic Processing of Health Care Products
Part 1: General Requirements Part 2: Filtration Part 3: Lyophilization Part 4: Clean-In-Place Technologies Part 5: Sterilization-In-Place Part 6: Isolator Systems
47

Aseptic Processing: Mrs Robyn Isaacson

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Aseptic Processing: Mrs Robyn Isaacson

Uploaded by

Copyright:

Available Formats

Aseptic Processing

Mrs Robyn Isaacson

Aseptic Processing - Overview

Two categories of sterile products

Aseptic Processing - Overview

Aseptic Processing - Overview

WHO GMP Grade A Grade B Grade C Grade D

US 209E M 3.5 M 3.5 M 5.5 M 6.5

ISO/TC (209) ISO 14644 ISO 5 ISO 5 ISO 7 ISO 8

EEC GMP Grade A Grade B Grade C Grade D

Grade C (equivalent to Class 10,000, ISO 7):

Grade B (equivalent to Class 100, ISO 5):

<3 10 100 200

Periodic health checks

Regular microbiological monitoring of operators

filtration should be carried out under positive pressure

heat tunnels often used for sterilization/depyrogenation of glass vials/bottles

tunnel supplied with HEPA filtered air

When filling from 5000-10000 units

When filling more than 10000 units

You might also like