Basic Principles of GMP

Sterile Pharmaceutical Products

Annex 6. TRS 902, 2002

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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

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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
9.1 9.6
false ceilings sealed
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Premises (continued)
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 (continued)
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 Pascals
Zone of greatest risk immediate environment
9.7 9.9

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Sterile Production
Premises (continued)
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
9.9 9.12

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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
treatment systems
10.1 10.5

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Equipment (continued)
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
10.6

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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
3.1 3.2

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Sterile Production
Sanitation (continued)
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
3.3

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Sterile Production
Sanitation (continued)
Limits of detection established
Alert and action, and monitoring trends of air quality
Table 1. Limits for microbial contamination (information only)

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3.4

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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Personnel (continued)
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

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8.4 8.6

Sterile Production
Personnel (continued)
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
not shedding fibres, and retain particles shed by
operators
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Personnel (continued)
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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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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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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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"
(That is "completed installation, equipment installed and
operating, but no operating personnel present")
4.1

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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
4.1
Clean areas for less critical operations
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Sterile Production
Air Classification System

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
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4.1 4.2.

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

4.1

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Processing
Minimize 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
4.15 4.16, 4.20 4.21

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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
4.17, 4.18, 4.28

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Processing
Water sources, water treatment systems and treated water
Monitored regularly
chemicals
biological contamination
endotoxin
Water specification
Records of results and action taken
4.19

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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
Gas used to purge solution or blanket a product passed
through a sterilizing filter
4.22 4.23

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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
4.26, 5.3

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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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Sterilization
Validation
all sterilization processes
special attention when non-pharmacopoeial 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
5.4 5.5
records maintained
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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
5.6 - 5.7

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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 the batch
release procedure
5.8 - 5.9

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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
6.2 6.3
sterilized cooling fluid/gas
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Sterile Production
Terminal Sterilization
Sterilization by moist heat (heating in an autoclave)
Water-wettable 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
6.4 6.6
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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)

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6.7

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
6.8 6.10
Radiation sensitive colour discs
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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
6.14 6.20

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Sterile Production
Terminal Sterilization
Monitoring of each cycle with biological indicators
time, pressure
temperature, humidity
gas concentration
Sterilization by gases and fumigants (2)
Post-sterilization storage controlled manner
ventilated conditions
defined limit of residual gas
validated process
Safety and toxicity issues

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6.21

Sterile Production
Terminally sterilized products

4.6 4.7
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Sterile Production
Terminally sterilized products

4.8 4.9

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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

4.10, 4.11, 4.14

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Sterile Production
Aseptic preparation

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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
each product
Batch processing records, sterility testing records, environmental
records should be reviewed
2.1 -2.2

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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
2.3

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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
11.1 11.3
results recorded
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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

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