Supplementary Training Modules on Good Manufacturing Practice

Sterile Pharmaceutical Products

Annex 6. TRS 902, 2002

Sterile

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

Sterile

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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, filling, sterilization, etc.
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1.1 1-2

Sterile Production
Premises  Design Avoid unnecessary entry of supervisors and control personnel Operations observed from outside  In clean areas, all exposed surfaces: Smooth, impervious, unbroken Minimize shedding and accumulation of particles, microorganisms Permit cleaning and disinfection No uncleanable recesses, ledges, shelves, cupboards, equipment Sliding doors undesirable False ceilings sealed 9.1 9.6
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Sterile Production
Premises
 In clean areas, all exposed surfaces (2):
Proper installation of pipes and ducts, no recesses, no unsealed openings Sinks and drains avoided, and excluded in Grade A and B areas Where installed, design, location, maintenance Effective cleanable traps Air breaks preventing backflow Floor channels open and easily cleanable
9.6.
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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
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9.7 9.9

Sterile Production
Premises  Pathogenic, highly toxic, radioactive materials  Pressure cascade may be different  Decontamination procedures air, equipment, 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.g. use of barriers
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9.9 9.12

Sterile Production
Equipment  Conveyer belts  Effective sterilization of equipment  Maintenance and repairs from outside the clean area If taken apart, resterilized before use Use clean instruments and tools  Planned maintenance, validation and monitoring Equipment, air filtration systems, sterilizers, water 10.1 10.5 treatment systems
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Sterile Production
Equipment  Water treatment plants and distribution system Design, construction, maintenance Operation and design capacity Testing programme

 Water for Injection (WFI) Produced, stored, distributed prevention of growth of microorganisms Constant circulation at temperature above 70, or not more than 4 degrees Celsius
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10.6

Sterile Production
Environmental Monitoring - I
Microbiological

 Air samples  Surface swabs  Personnel swabs

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Sterile Production
Environmental Monitoring - II Physical  Particulate matter  Differential pressures  Air changes, airflow patterns  Clean-up time/recovery  Filter integrity  Temperature and relative humidity  Airflow velocity
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Sterile Production
Sanitation  Frequent, 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, stored for defined periods of time Sterilized before use, when used in Grade A or B areas
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3.1 3.2

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, surface sampling (swabs and contact plates) Sampling methods should not contaminate the area  Results considered when batch release is done
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3.3

Sterile Production
Sanitation
 Limits of detection established  Alert and action, and monitoring trends of air quality
Table 1. Limits for microbial contamination (Information only)
Grade Air sample (CFU/m3) Settle plates (90mm diameter) (CFU/4hours) A B C D
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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

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, hygiene, microbiology  Special cases Supervision in case of outside staff 8.1 8.3 Decontamination procedures (e.g. staff who worked with animal tissue materials)
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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, jewellery and cosmetics
8.4 8.6

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Sterile Production
Personnel  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 no fibres to be shed 8.7 Grade A and B Headgear, beard and moustache covered, masks, gloves No shedding of fibres, and retain particles shed by operators
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Sterile Production
Personnel  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  Disinfect gloves during operations  Washing of garments separate laundry facility  No damage, and according to validated procedures
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8.8 8.9

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, their purpose and air classification
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Sterile Production
Possible Issues  Poor design of the building  Poor design of the systems, e.g. water, HVAC  Flow of personnel  Flow of material  No validation or qualification  Old facilities not complying with current requirements

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

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Sterile Production
Two categories of manufacturing operations:

 Terminally sterilized
prepared, filled and sterilized

 Aseptic preparation
some or all stages
1.3
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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").
4.1
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Sterile Production
Manufacture of sterile preparations  For sterile pharmaceutical preparations:  Grade A Local zone, high risk operations, e.g. filling, 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
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4.1

Sterile Production
Air Classification System
Grade At rest 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 In operation

3.1

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Sterile Production
Manufacture of sterile preparations  To reach Grade B, C and D, the number of air changes should be appropriate to the size of the area, number of personnel, 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.g. EU, ISO, Japan, USA 4.1 4.2.
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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.g. validation runs, media fills, environment, time limits based on microbiological contamination/bioburden found)

4.3 4.5

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Sterile Production
Airborne particulate classification

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 5 ISO 5 ISO 7 ISO 8

EEC GMP Grade A Grade B Grade C Grade D

4.1

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Sterile Production
Processing  Minimise contamination all stages including before sterilization and during processing  No unsuitable materials, e.g. live microbiological organisms  Minimize activities staff movement controlled and methodical avoid shedding of particles  Temperature and humidity comfortable  Containers and materials in the area
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4.15 4.16, 4.20 4.21

Sterile Production
Processing  Validation should not compromise the processes  Aseptic process validation: Sterile media fill (broth fills) Simulate actual operation intimate as closely as possible Simulate worst expected condition Use appropriate medium/media Sufficient number of units - e.g. 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
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4.17, 4.18, 4.28

Sterile Production
Processing  Water sources, water treatment systems and treated water  Monitored regularly Chemicals Biological contamination Endotoxin  Water specification  Records of results and action taken
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4.19

Sterile Production
Processing  Components, 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, 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
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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
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4.26, 5.3

Sterile Production
Processing  Time intervals: Components, bulk containers, 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
4.23 - 4.24
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Sterile Production
Group session 2  Considering the same factory as in the previous group session, discuss the process of sterilization.  List all the items that will need to be sterilized (and indicate the choice of sterilization process).  What are the key features you should find in each sterilization situation?  Discuss the relevance, need, and the extent of qualification and validation required.

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Sterile Production
Possible Issues  Autoclave - no pressure gauge  Autoclave - no temperature recorder  Autoclave - superheated steam  Clean room - pressure differentials  Exposure for settle plates  Interlocks turned off  Rusty Laminar airflow cabinets  HEPA filters not checked regularly
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Sterile Production
Sterilization
 Methods of sterilization:
Moist or dry heat Irradiation (ionizing radiation) Sterilizing gaseous agents (e.g. ethylene oxide) Filtration with subsequent aseptic filling

 Whenever possible: Terminal sterilization by heat in their final container - method of choice
5.1 5. 2
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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
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5.4 5.5

Sterile Production
Sterilization
 For effective sterilization:  Whole of the material subjected to the treatment  Biological indicators:  Additional method of monitoring  Storage and use, quality checked through positive control  Risk of contamination
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5.6 - 5.7

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 5.8 - 5.9 the batch release procedure
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Sterile Production
Terminal Sterilization
 Sterilization by heat  Sterilization by moist heat  Sterilization by dry heat  Sterilization by radiation  Sterilization by gases and fumigants
6
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Sterile Production
Terminal Sterilization Sterilization by heat  Recording of each cycle, e.g. 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
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6.2 6.3

Sterile Production
Terminal Sterilization Sterilization by moist heat (heating in an autoclave)  Water-wetable materials only, and aqueous formulations  Temperature, 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
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6.4 6.6

Sterile Production
Terminal Sterilization Sterilization by dry heat  For non-aqueous liquids, 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
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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, 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
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6.8 6.10

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 6.10 6.13
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Sterile Production
Terminal Sterilization Sterilization by gases and fumigants  Only when no other method is suitable  e.g. ethylene oxide, hydrogen peroxide vapour  Validation: Also prove the gas has no damaging effect on product  Time and conditions for degassing (specified limits) - 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, pressure temperature, humidity and gas concentration
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6.14 6.20

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

6.21
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Sterile Production
Terminally sterilized products
Grade D Preparation Components and product Remark Ensure low microbial and particulate count e.g. product actively supports microbial growth, or is held for a long period of time before sterilization, or is not prepared mainly in closed containers -

Product at unusual risk of microbial contamination

Filling before sterilization

4.6 4.7
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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. slow filling operation, or Wide neck containers, or Exposure for a few seconds before sealing Ointments, creams, suspensions, emulsions

4.8 4.9

Sterile

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Sterile Production
Aseptic processing and sterilization by filtration
Aseptic processing  Objective is to maintain the sterility of a product, assembled from sterile components  Operating conditions so as to prevent microbial contamination  What do you think are the aspects that require careful attention? 7.1 7.2
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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
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7.3

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, creams, suspensions, emulsions Remark

A (in B background)

When the product is exposed and filtered

4.10, 4.11, 4.14

Sterile

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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)

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Preparation of solutions (if not to be sterile filtered later) Handling and filling of aseptically prepared products, Handling of exposed sterile equipment Transfer of partially closed containers, before complete stoppering
January 2006

e.g. in freeze drying (Grade B environment if in sealed transfer trays)

4.10 4.13

Sterile Production
Sterilization by filtration  Through a sterile filter of 0,22 m or less, 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, just before filling - no fibre shedding or asbestos filters  Filter integrity testing immediately after use also before use if possible
7.4 7.7
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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, other filters at appropriate intervals
7.7
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Sterile Production
Sterilization by Filtration (3)
 Same filter not used for more than one working day, unless validated  No filter interaction with product, e.g.
removal of ingredients releasing substances into product

7.8 7.9
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Sterile Production
Quality Control  Samples for sterility testing should be representative  From parts of the batch, most at risk Aseptic filling at beginning and end of batch filling, 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, and validated for each2.1 -2.2 product  Batch processing records, sterility testing records, environmental records should be reviewed

Sterile

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Sterile Production
Quality Control
 Endotoxin testing for injectable products
Water for injection, intermediate and finished product

 Always for large volume infusion solutions  Pharmacopoeia method, validated for each product  Failure of the test investigation  Corrective action
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2.3

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, and equipment performance checked at intervals results recorded
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11.1 11.3

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

Sterile

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January 2006