Supplementary Training Modules on Good Manufacturing Practice

Sterile Pharmaceutical Products

Annex 6. TRS 902, 2002

Sterile

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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
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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

Sterile

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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

Sterile

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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

Sterile

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

Temperature/humidity

Sterile

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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

Sterile

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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.

Sterile

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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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January 2006

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
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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