Supplementary Training Modules on Good Manufacturing Practice

Sterile Pharmaceutical Products

Annex 6. TRS 902, 2002



Slide 1 of 62

January 2006

Sterile Production
Objectives  To review basic GMP requirements in the manufacture of sterile pharmaceutical products  To review air classifications for activities related to the manufacture of sterile products  To review the different types of sterilization methods  To review quality assurance aspects in the manufacture and control of sterile products  To consider current issues applicable in your country
Sterile | Slide 2 of 62 January 2006

Sterile Production
GMP Requirements for Sterile Products 
Additional rather than replacement  Specific points relating to minimizing risks of contamination
± microbiological ± particulate matter ± pyrogen



Slide 3 of 62

January 2006

Sterile Production General Considerations  Production in clean areas  Appropriate standard of cleanliness  Filtered air supplied  Airlocks for entry ± Personnel and/or equipment ± Materials  Separate areas for operations ± component preparation (containers and closures) ± product preparation. sterilization. filling. etc.1 ± 1-2 . Sterile | Slide 4 of 62 January 2006 1.

6 Sterile | Slide 5 of 62 January 2006 .1 ± 9. impervious. ledges.Sterile Production Premises  Design ± Avoid unnecessary entry of supervisors and control personnel ± Operations observed from outside  In clean areas. all exposed surfaces: ± Smooth. equipment ± Sliding doors undesirable ± False ceilings sealed 9. unbroken ± Minimize shedding and accumulation of particles. microorganisms ± Permit cleaning and disinfection ± No uncleanable recesses. cupboards. shelves.

6. Sterile | Slide 6 of 62 January 2006 . design. location.Sterile Production Premises  In clean areas. no unsealed openings ± Sinks and drains avoided. maintenance ‡ Effective cleanable traps ‡ Air breaks preventing backflow ‡ Floor channels open and easily cleanable 9. no recesses. and excluded in Grade A and B areas ‡ Where installed. all exposed surfaces (2): ± Proper installation of pipes and ducts.

7 ± 9.Sterile Production Premises  Changing rooms ± Designed as airlocks ± Effective flushing with filtered air ± Separate rooms for entry and exit desirable ± Hand washing facilities ± Interlocking system for doors ± Visual and/or audible warning system  Use filtered air supply to maintain pressure cascade  Pressure differential approximately 10 to 15 Pascales  Zone of greatest risk ± immediate environment Sterile | Slide 7 of 62 January 2006 9.9 .

g. highly toxic.9 ± 9. garments  Qualification including airflow patterns ± No risk to the product  Warning system to indicate failure in air supply  Pressure indicators ± results regularly recorded  Restricted access ± e. equipment.12 . use of barriers Sterile | Slide 8 of 62 January 2006 9.Sterile Production Premises  Pathogenic. radioactive materials  Pressure cascade may be different  Decontamination procedures ± air.

Sterile Production Equipment  Conveyer belts  Effective sterilization of equipment  Maintenance and repairs from outside the clean area ± If taken apart.5 treatment systems Sterile | Slide 9 of 62 January 2006 . air filtration systems. water 10. sterilizers.1 ± 10. resterilized before use ± Use clean instruments and tools  Planned maintenance. validation and monitoring ± Equipment.

maintenance ± Operation and design capacity ± Testing programme  Water for Injection (WFI) ± Produced. construction. distributed ± prevention of growth of microorganisms ± Constant circulation at temperature above 70. stored.Sterile Production Equipment  Water treatment plants and distribution system ± Design.6 . or not more than 4 degrees Celsius Sterile | Slide 10 of 62 January 2006 10.

I Microbiological  Air samples  Surface swabs  Personnel swabs Sterile | Slide 11 of 62 January 2006 .Sterile Production Environmental Monitoring .

airflow patterns  Clean-up time/recovery  Filter integrity  Temperature and relative humidity  Airflow velocity Sterile | Slide 12 of 62 January 2006 .II Physical  Particulate matter  Differential pressures  Air changes.Sterile Production Environmental Monitoring .

1 ± 3. thorough cleaning of areas necessary  Written programme  Regular monitoring to detect resistant strains of microorganisms  Chemical disinfection  Monitoring of disinfectants and detergents  Dilutions ± Clean containers.2 . when used in Grade A or B areas Sterile | Slide 13 of 62 January 2006 3. stored for defined periods of time ± Sterilized before use.Sterile Production Sanitation  Frequent.

surface sampling (swabs and contact plates) ± Sampling methods should not contaminate the area  Results considered when batch release is done Sterile | Slide 14 of 62 January 2006 3.Sterile Production Sanitation  Monitoring of clean areas  Monitoring of personnel and surfaces after critical operations  Frequent monitoring in areas where aseptic operations are carried out ± Settle plates. volumetric air samples.3 .

Limits for microbial contamination (Information only) Grade Air sample (CFU/m3) Settle plates (90mm diameter) (CFU/4hours) A B C D Sterile | Slide 15 of 62 3.4 Contact plates (55mm diameter) (CFU/plate) <3 5 25 50 Glove print (5 fingers) (CFU/glove) <3 5 - <3 10 100 200 January 2006 <3 5 50 100 . and monitoring trends of air quality Table 1.Sterile Production Sanitation  Limits of detection established  Alert and action.

3 ± Decontamination procedures (e. hygiene.g. staff who worked with animal tissue materials) Sterile | Slide 16 of 62 January 2006 . microbiology  Special cases ± Supervision in case of outside staff 8.1 ± 8.Sterile Production 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.

6 Sterile | Slide 17 of 62 January 2006 . jewellery and cosmetics 8.4 ± 8.Sterile Production Personnel  High standards of hygiene and cleanliness  Periodic health checks  No shedding of particles  No introduction of microbiological hazards  No outdoor clothing  Changing and washing procedure  No watches.

shoes ‡ no fibres to be shed 8.Sterile Production Personnel  Clothing of appropriate quality: ± Grade D ‡ hair. beard and moustache covered. high neck). masks. and retain particles shed by operators Sterile | Slide 18 of 62 January 2006 . moustache covered ‡ Single or 2-piece suit (covering wrists.7 ± Grade A and B ‡ Headgear. beard. gloves ‡ No shedding of fibres. moustache covered ‡ Protective clothing and shoes ± Grade C ‡ Hair. beard.

and according to validated procedures Sterile | Slide 19 of 62 January 2006 8. or once a day (if supportive data)  Change gloves and masks at every working session  Disinfect gloves during operations  Washing of garments ± separate laundry facility  No damage.8 ± 8.Sterile Production Personnel  Outdoor clothing not in change rooms leading to grade B and C rooms  Change at every working session.9 .

their purpose and air classification Sterile | Slide 20 of 62 January 2006 .Sterile Production Group session 1  You are asked to visit a factory producing the following product lines: ± Injections in ampoules and vials. including insulin. vaccines and heat-stable pharmaceuticals ± Sterile eye ointment  Describe the type of facility you would expect to find  List the typical rooms.

HVAC  Flow of personnel  Flow of material  No validation or qualification  Old facilities not complying with current requirements Sterile | Slide 21 of 62 January 2006 . e.Sterile Production Possible Issues  Poor design of the building  Poor design of the systems. water.g.

Sterile Production
Possible Issues (continued) 
Particulate levels/microorganisms  Differential pressures  Air changes  Temperature/humidity



Slide 22 of 62

January 2006

Sterile Production
Two categories of manufacturing operations: 

Terminally sterilized
± prepared, filled and sterilized 

Aseptic preparation
± some or all stages
Sterile | Slide 23 of 62 January 2006

Sterile Production
Manufacture of sterile preparations  Classification of clean areas  Manufacturing operation in an appropriate environment cleanliness level  Minimize risks ± particulate and microbiological contamination ± product and material  Meet classification "at rest" ± (i.e. "completed installation, equipment installed and operating, but no operating personnel present").
Sterile | Slide 24 of 62 January 2006

g. e. high risk operations.Sterile Production Manufacture of sterile preparations  For sterile pharmaceutical preparations:  Grade A ± Local zone. filling.1 . aseptic connections ± Usually UDAF systems used  Grade B ± Background environment to grade A (in case of aseptic preparation and filling)  Grade C and Grade D ± Clean areas for less critical operations Sterile | Slide 25 of 62 January 2006 4.

5 .5.5 .Sterile Production Air Classification System Grade At rest maximum permitted number of particles/m3 0.5.0 µm A B C D 3 500 3 500 350 000 3 500 000 > 5 µm 0 0 2 000 20 000 0.1 Sterile | Slide 26 of 62 January 2006 .0 µm 3 500 350 000 3 500 000 not defined >5µ 0 2 000 20 000 not defined In operation 3.

number of personnel.Sterile Production Manufacture of sterile preparations  To reach Grade B. Sterile | Slide 27 of 62 January 2006 . USA 4.1 ± 4.g. Japan. EU. C and D.2. the number of air changes should be appropriate to the size of the area. ISO. equipment present  Minimum of 20 air changes per hour  Clean up time about 15-20 minutes  Good airflow pattern in the area  HEPA-filtered air  Suitable methods to determine particulate matter and micro± e.

Sterile Production Manufacture of sterile preparations  Control particulate during operation  Monitoring during operation  Alert and action limits for particulate and micro  Action taken when exceeded  Area grades should be proven (e. time limits ± based on microbiological contamination/bioburden found) 4. environment. validation runs.3 ± 4.g. media fills.5 Sterile | Slide 28 of 62 January 2006 .

Sterile Production Airborne particulate classification WHO GMP Grade A Grade B Grade C Grade D US 209E M 3.1 Sterile | Slide 29 of 62 January 2006 .5 M 6.5 US Customary Class 100 Class 100 Class 10 000 Class 100 000 ISO/TC (209) ISO 5 ISO 5 ISO 7 ISO 8 EEC GMP Grade A Grade B Grade C Grade D 4.5 M 5.5 M 3.

live microbiological organisms  Minimize activities ± staff movement controlled and methodical ± avoid shedding of particles  Temperature and humidity comfortable  Containers and materials in the area Sterile | Slide 30 of 62 January 2006 4. e.g.Sterile Production Processing  Minimise contamination ± all stages including before sterilization and during processing  No unsuitable materials.16. 4.21 .20 ± 4.15 ± 4.

28 .17.e. equal to batch size (small batches) ‡ acceptable limit ‡ investigations ± Revalidation: periodic and after change  New processing procedures validated ± Revalidation after significant changes ± And regular intervals Sterile | Slide 31 of 62 January 2006 4.Sterile Production Processing  Validation ± should not compromise the processes  Aseptic process validation: Sterile media fill (³broth fills´) ± Simulate actual operation ± intimate as closely as possible ± Simulate worst expected condition ± Use appropriate medium/media ± Sufficient number of units .g.18. 4. 4.

water treatment systems and treated water  Monitored regularly ± Chemicals ± Biological contamination ± Endotoxin  Water specification  Records of results and action taken Sterile | Slide 32 of 62 January 2006 4.Sterile Production Processing  Water sources.19 .

bulk product containers and equipment ± fibre generation ± no recontamination after final cleaning ± stage properly identified ± sterilized when used in aseptic areas  Used in clean areas.Sterile Production Processing  Components. passed through double-ended sterilizers or use triple wrapping 4.22 ± 4.23  Gas used to purge solution or blanket a product ± passed through a sterilizing filter Sterile | Slide 33 of 62 January 2006 .

3 . 5.26.Sterile Production Processing  Bioburden monitored ± Products: Before sterilization ± Working limits established ± Solutions to be filtered before filling (especially LVP) ± Pressure release outlets ± hydrophobic microbiological air filters  Starting materials ± microbiological contamination should be minimal  Monitored as per specification Sterile | Slide 34 of 62 January 2006 4.

equipment  Washing and drying and sterilization. and sterilization and use ± As short as possible ± Time limit validated  Time intervals: Product  Start of preparation of solution and sterilization (filtration) ± As short as possible ± Maximum time set for each product 4.4.23 . bulk containers.Sterile Production Processing  Time intervals: Components.24 Sterile | Slide 35 of 62 January 2006 .

and the extent of qualification and validation required.  List all the items that will need to be sterilized (and indicate the choice of sterilization process). need. discuss the process of sterilization.  What are the key features you should find in each sterilization situation?  Discuss the relevance.Sterile Production Group session 2  Considering the same factory as in the previous group session. Sterile | Slide 36 of 62 January 2006 .

no temperature recorder  Autoclave .no pressure gauge  Autoclave .pressure differentials  Exposure for settle plates  Interlocks turned off  Rusty Laminar airflow cabinets  HEPA filters not checked regularly Sterile | Slide 37 of 62 January 2006 .superheated steam  Clean room .Sterile Production Possible Issues  Autoclave .

ethylene oxide) Filtration with subsequent aseptic filling  Whenever possible: Terminal sterilization by heat in their final container .method of choice 5.Sterile Production Sterilization  Methods of sterilization: ± ± ± ± Moist or dry heat Irradiation (ionizing radiation) Sterilizing gaseous agents (e. 2 Sterile | Slide 38 of 62 January 2006 .1 ± 5.g.

Sterile Production Sterilization  Validation ± All sterilization processes ± Special attention when non-pharmacopoeia methods are used ± Non-aqueous or oily solutions  Before the method is adopted ± its suitability and efficacy demonstrated with desired conditions: ± All parts of the load ± Each type of load ± Physical measurements and biological indicators (where appropriate) ± Verified at least annually and after change ± Records maintained Sterile | Slide 39 of 62 January 2006 5.5 .4 ± 5.

Sterile Production Sterilization  For effective sterilization:  Whole of the material subjected to the treatment  Biological indicators:  Additional method of monitoring  Storage and use.5. quality checked through positive control  Risk of contamination Sterile | Slide 40 of 62 January 2006 5.7 .6 .

8 .5.9 the batch release procedure Sterile | Slide 41 of 62 January 2006 .Sterile Production Sterilization  Differentiation between sterilized and not-yet-sterilized products  Each basket/tray or other carrier. properly labelled ± Name of material ± Batch number ± Sterilization status  Use of autoclave tape  Sterilization records for each run ± approved as part of 5.

Sterile Production Terminal Sterilization  Sterilization by heat  Sterilization by moist heat  Sterilization by dry heat  Sterilization by radiation  Sterilization by gases and fumigants 6 Sterile | Slide 42 of 62 January 2006 .

e.Sterile Production Terminal Sterilization Sterilization by heat  Recording of each cycle. time and temperature chart ± Temperature: validated coolest part ± Check from second independent probe ± Additional chemical or biological indicators  Heating phase: Sufficient time for the whole load ± Determined for each load  Cooling phase: After sterilization cycle ± Precautions to prevent contamination ± Sterilized cooling fluid/gas Sterile | Slide 43 of 62 January 2006 6.2 ± 6.3 .g.

and aqueous formulations  Temperature.4 ± 6. time and pressure monitored  Temperature recorder independent of the controller  Independent temperature indicator  Drain ± temperature recorded from this position  Regular leak test when vacuum is part of the cycle  Material allows for removal of air and penetration of steam  All parts of the load in contact with steam  Quality of the steam ± no contamination Sterile | Slide 44 of 62 January 2006 6.6 .Sterile Production Terminal Sterilization Sterilization by moist heat (heating in an autoclave)  Water-wetable materials only.

dry powders  Air circulation in the chamber  Positive pressure in chamber to prevent entry of non-sterile air  HEPA filtered air supplied  When removing pyrogens. challenge tests ± validation (using endotoxins) 6.7 Sterile | Slide 45 of 62 January 2006 .Sterile Production Terminal Sterilization Sterilization by dry heat  For non-aqueous liquids.

10 . responsibilities  Measurement of dose during procedure  Dosimeters independent of dose rate ± quantitative measurement ± number. location and calibration time-limit  Biological indicators only as additional control  Radiation sensitive colour discs Sterile | Slide 46 of 62 January 2006 6.Sterile Production Terminal Sterilization Sterilization by radiation  Suitable for heat-sensitive materials and products ± confirm suitability of method for material ± ultraviolet irradiation not acceptable  Contracting service ± ensure validation status.8 ± 6.

Sterile Production Terminal Sterilization Sterilization by radiation (2)  Information forms part of the batch record  Validation to cover effects of variation in density of packages  Handling procedures to prevent misidentification of irradiated and non-irradiated materials  Each package to have a radiation-sensitive indicator  Total radiation dose administered within a predetermined period of time 6.13 Sterile | Slide 47 of 62 January 2006 .10 ± 6.

20 . hydrogen peroxide vapour  Validation: Also prove the gas has no damaging effect on product  Time and conditions for degassing (specified limits) . ethylene oxide. humidity and gas concentration Sterile | Slide 48 of 62 January 2006 6.g.residue  Direct contact with microbial cells essential ± Nature and quantity of packaging materials  Humidity and temperature equilibrium  Monitoring of each cycle with biological indicators ± time.Sterile Production Terminal Sterilization Sterilization by gases and fumigants  Only when no other method is suitable  e.14 ± 6. pressure ± temperature.

21 Sterile | Slide 49 of 62 January 2006 .Sterile Production Terminal Sterilization Sterilization by gases and fumigants (2)  Post-sterilization storage ± controlled manner ± Ventilated conditions ± Defined limit of residual gas ± Validated process  Safety and toxicity issues 6.

7 Sterile | Slide 50 of 62 January 2006 .Sterile Production Terminally sterilized products Grade D Preparation Components and product Remark Ensure low microbial and particulate count e. or is held for a long period of time before sterilization.g. or is not prepared mainly in closed containers - C Product at unusual risk of microbial contamination C Filling before sterilization 4.6 ± 4. product actively supports microbial growth.

or Wide neck containers.8 ± 4. slow filling operation.Sterile Production Terminally sterilized products Grade A in C background Preparation Filling before sterilization if product at unusual risk of contamination from environment Preparation and filling Remark e.g. creams. emulsions C 4.9 Sterile | Slide 51 of 62 January 2006 . or Exposure for a few seconds before sealing Ointments. suspensions.

Sterile Production Aseptic processing and sterilization by filtration Aseptic processing  Objective is to maintain the sterility of a product.1 ± 7. assembled from sterile components  Operating conditions so as to prevent microbial contamination  What do you think are the aspects that require careful attention? 7.2 Sterile | Slide 52 of 62 January 2006 .

3 .Sterile Production Aseptic processing and sterilization by filtration Aseptic processing (2)  Careful attention to:  Environment  Personnel  Critical surfaces  Container/closure sterilization  Transfer procedures  Maximum holding period before filling Sterile | Slide 53 of 62 January 2006 7.

emulsions Remark A (in B background) When the product is exposed and filtered 4. 4. creams. 4.10.14 Sterile | Slide 54 of 62 January 2006 . suspensions.11.Sterile Production Aseptic preparation Grade D C Preparation Components after washing Preparation of solutions to be sterile filtered later in the process Preparation and filling of sterile ointments.

g.10 ± 4.Sterile Production Aseptic preparation Grade A (in B background) Preparation Sterile starting materials and components Remark (Unless subjected to sterilization or filtration through a microorganism retaining filter later in the process) A (in B background) A (in B background) A (in B background) A (in B background) Sterile | Slide 55 of 62 Preparation of solutions (if not to be sterile filtered later) Handling and filling of aseptically prepared products. before complete stoppering January 2006 e.13 . Handling of exposed sterile equipment Transfer of partially closed containers. in freeze drying (Grade B environment if in sealed transfer trays) 4.

7 Sterile | Slide 56 of 62 January 2006 .no fibre shedding or asbestos filters  Filter integrity testing immediately after use ± also before use if possible 7.22 µm or less. just before filling .Sterile Production Sterilization by filtration  Through a sterile filter of 0. into previously sterilized containers ± remove bacteria and moulds ± not all viruses or mycoplasmas  Consider complementing with some degree of heat treatment  Double filter layer or second filtration advisable.4 ± 7.

other filters at appropriate intervals 7.Sterile Production Sterilization by Filtration (2)  Validation to include ± Time taken to filter a known volume ± Pressure difference to be used across the filter  Significant differences to be noted and investigated. recorded in batch records  Integrity of gas and air vent filters checked after use.7 Sterile | Slide 57 of 62 January 2006 .

Sterile Production Sterilization by Filtration (3)  Same filter not used for more than one working day.8 ± 7. e. unless validated  No filter interaction with product. ± removal of ingredients ± releasing substances into product 7.9 Sterile | Slide 58 of 62 January 2006 .g.

Sterile Production Quality Control  Samples for sterility testing should be representative  From parts of the batch. and after interruptions ± Heat sterilized ± coolest part of the load  Sterility of the batch ensured through validation ± Validated sterilization cycle ± Media fill  Sterility test procedure as per pharmacopoeia.1 -2. and validated for each2. environmental records should be reviewed Sterile | Slide 59 of 62 January 2006 . sterility testing records.2 product  Batch processing records. most at risk ± Aseptic filling ± at beginning and end of batch filling.

validated for each product  Failure of the test ± investigation  Corrective action Sterile | Slide 60 of 62 January 2006 2.Sterile Production Quality Control  Endotoxin testing for injectable products ± Water for injection. intermediate and finished product  Always for large volume infusion solutions  Pharmacopoeia method.3 .

and equipment performance checked at intervals ± results recorded Sterile | Slide 61 of 62 January 2006 11.1 ± 11.Sterile Production Finishing of products  Containers closed by means of validated methods  Samples checked for integrity  Maintenance of vacuum (where applicable) checked  Parenteral products inspected individually  Visual inspection under suitable and controlled conditions: ± illumination and background ± eyesight checks of operators ± allowed frequent breaks  Other methods: ± validated.3 .

acceptance criteria and frequency of testing. devise a plan for monitoring of the facility.Sterile Production Group session 3  Considering the same factory as in the previous group sessions. Sterile | Slide 62 of 62 January 2006 .  List the parameters to be tested. tests to be used.

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