Technical Considerations: Steam Sterilisation

QT/ST/01

PURPOSE:

To provide a document to detail various aspects required to be known prior to use of

steam steriliser.

SCOPE:

Applicable to steam sterilisation of items other than terminal sterilisation of product.

RESPONSIBILITY:

All Staff associated with steam sterilisation

DEFINITION:

A validated process used to render a product free of viable organisms.

HEALTH, SAFETY AND ENVIRONMENT:

1. When removing items from the autoclave, wear a rubber apron, rubber sleeve
protectors, heat resistant gloves and a face shield. Remove the load and let
the glassware cool for 15 minutes before touching with ungloved hands
2. Liquid loads should stand in an out-of-the-way place for a full hour before
touching with ungloved hands
3. Before loading containers of liquid into the autoclave the caps must be
loosened to avoid having the bottles shattered during pressurisation.
4. The drain stainer should be clean before loading the autoclave.
5. Instead of sterilising empty bottles or flasks add ¼ to ½ inch of water so the
bottles will heat evenly.
6. Use a tray with a solid bottom and walls to contain the bottles and catch
spills.
7. Do not load plastic materials that are not compatible with the autoclave.
8. Always Operate the Autoclave as per latest SOP

PROCEDURE:

This part details various technical aspects that one should know before using steam
steriliser.

TECHNICAL CONSIDERATIONS:

Type of cycle:

Over kill sterilisation approach:

A cycle that provides a minimum 12-log reduction of a resistant biological indicator
with a known D-value of not less than 1 minute

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Load Classification:

Porous loads ( Hard Goods):

Loads of materials for which the contaminant microbial populations are inactivated
through direct contact with the steam supplied to the autoclave.

Examples of Porous loads include but are not restricted to:

I. Filters (Disc, membranes, cartridge membranes and depth filters)
II. Stoppers, plugs and wads ( rubber and polymeric materials)
III. Tubing and hoses
IV. Garments ( packed fabrics)
V. Cleaning equipment
VI. Machine Change parts
VII. SS vessels

Materials constituting porous loads may be presented to the autoclave in a variety of

ways.
Examples include but are not restricted to:
• Steam and air permeable wrappings ( paper or other polymeric wrapping
materials, non shedding fabric or combinations )
• In closed but not sealed boxes( stainless steel or anodised aluminium and
perforated, to allow for steam penetration, air removal and drainage of any
steam condensate which may accumulate)
• Open trays ( with or without steam and air permeable wrapping)
• In static or moving containers i.e stoppers

Following factors should be taken into account when assembling loads for
sterilisation by saturated steam:
• Penetration of the load by steam and effective removal of air and condensate
• Protection of items against contamination from microorganisms and
particulate matter before, during and after autoclaving.
• Prevention of localised super heating
• Prevention of wetting from condensate caused by ineffective drainage

To meet these requirements:

Loading configurations should be documented, and only authorised loads should be
used. Schematic loading diagrams should be included in documentation and be
available for reference. Loading patterns within tins or containers should also be
documented.
Load configurations should ensure that flat surfaces such as the outside of metal
boxes and trays, do not come into contact with the interior surfaces of the chamber.
Contact between flat surfaces of metal boxes and trays is minimised by use of racks
with perforated, and, if necessary, adjustable shelving.

Ensure in porous loads that:

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• All Air is removed from the autoclave chamber and load prior to the
sterilisation hold phase of the cycle
• All Air and non condensable gases are removed from the steam supply to the
autoclave
• Steam supplied to the autoclave is saturated ( correct dryness factor)
• Steam does not contain substances that would chemically contaminate the
material it comes into contact with and subsequently product

Non Porous loads ( Fluid loads ):

Non porous loads are loads of closed containers in which aqueous loads are
terminally sterilised. The sterilisation of container contents is achieved through
transfer of heat from the steam to the container exterior and then to the aqueous
liquid contained within.

Examples of Non Porous loads include but are not restricted to:
I. Formulations ( Solutions, Suspensions and or Emulsions)
II. In-process aqueous fluids in glass jars or containers
III. Laboratory media or aqueous fluids for use in sterility testing and other
microbiological work
IV. Pos t test or post process fluids in rigid glass

Non porous loads are ordinarily arranged to maximise the penetration of steam to
the load
Ensure in non porous loads that
• The load is positioned in the same position during validation and routine
processing
• That the temperature across the load is uniform to avoid inadvertent over or
under processing
• That the bioburden content of the filled containers confirm to that allowed for
during cycle development
• That the air overpressure (where utilised) is sufficient to minimise the
breakage or distortion of containers.

GENERAL INSTRUCTIONS:

1. Aluminium foil ,glassine paper and other non permeable materials must not
be used for wrapping items to be sterilised by Autoclaving.
2. Bags used to wrap material for sterilisation should not shed particles in the
area and be steam permeable.
3. Flexible tubing used for connections should not be greater than three meters
in length
4. Number of coils that should be used for tubing is not more than three coils
while loading in the steriliser. Shorter tubes should be kept in U shape.
5. There should be no obstruction at the open ends of the tubing to ensure
steam penetration across tubing
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6. All screw capped bottles should be left crack open i.e. one fourth turn to
finger tightening during sterilisation
7. Vent should be open or Gap should be maintained between the pressure
vessel and lid during sterilisation to avoid pressure built up during pulsing
cycle.
8. Aluminium foil that is needed for covering items in filling area should be
sterilised by cutting in pieces and keeping loosely in perforated trays. Not
more than three pieces to be kept in one tray.
9. Folding and packing of garments should be same as done during validation
for all garments put for sterilisation. SOP for the same should be in place.
10.Plastic, rubber and other heat sensitive items should not be loaded in the
autoclave
11.Stainless steel items can be safely put in the porous load cycle of Autoclave
12.Best mode of sterilisation of glass is dry heat steriliser.
13.All items left open in the steriliser should be immediately closed on opening
the Autoclave.
14.Loading should be done from unclean side of Autoclave and unloading
should be done from sterile side of autoclave for aseptic areas.
15.Items once unloaded should be left under LAF station till use for a period
ascertained in validation.
16.Loading should be done strictly as per Load pattern fixed during Autoclaving.
17.Autoclave cycles should be same as those specified in SOP after validation.

PRECAUTIONS:

Certain precautions to be taken prior to commenc ing Autoclave cycles

I. Check that the adequate stationary is available for printing.
II. Check that ink recorder is in working condition.
III. Check PLC is operational if applicable.
IV. Check steam condensate is drained off.
V. Check drain is free and not choked.
VI. Ensure there is no water in the autoclave chamber.
VII. Steriliser is cleaned as per routine schedule and records
maintained.
VIII. Ensure all dials and gauges are calibrated.
IX. Compressed air and steam lines are operational

PULSING CYCLES:

Vacuum pulsing system employ a series of alternating steam pulses followed by

vacuum excursions. Steam pulses serve to rapidly fill the voids forced by the vacuum
pulses, causing incremental decreases in the amount of residual air. The number of
pulses may vary and upto seven cycles may be necessary to adequately remove the

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air.These cycles are used to sterilise loads that contain items from which air removal
can be difficult.
Examples of such load items include:
Coiled hoses,Filter housings,densly packed containers of stoppers and packages of
clean room garments.

DRYING CYCLES:

In certain situations, wet components post streilisation may adversly affect the finish
product quality.In these instances a drying cycle is employed.The drying cycle
follows the sterilisation dwell phase
Options that can be employed for drying:
Heating: A reheating of the items in the chamber using the vessel jacket to
evaporate moisture over an extended period.
Deep Vacuum:Vacuum is drawn on the chamber and maintained for an extended
period to enhance evaporation
Pulsing: Alternating pulses of vacuum and compressed air are utilised.The
introduction of dry air increases the evaporation rate in subsequent vacuum.

THERMOCHEMICAL INDICATOR:

A thermochemical indicator is a device, which responds to sterilisation proc ess

parameters in a non quantitative fashion. These types of devices are placed on the
outside of containers or other sterilised units to verify that the product or load has
been exposed to a sterilisation cycle in order to prevent mix up of sterilised and non
sterilised products. Typical example is autoclave tape that indicates by a change in
colour that the product has been exposed to a certain temperature but does not
identify how long that condition was experienced.

STERILISATION CYCLE:

Optimal Sterilisation parameters:

Chamber come-up time
Load come-up time
Exposure time
Exposure temperature
Pressure requirements
Equilibration time
Drain or chamber temperature
Time allowed for heat-up and cooling
Temperature distribution
Heat penetration

Chamber Come up time: Elasped time from from the initiation of the cycle to the
beginning of the dwell or exposure phase of the sterilisation cycle.

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Load Come up Time: The time for slowest to heat item in the load to reach or exceed
the steriliser set point temperature.

Exposure time: The time at which the autoclave is held at the desired operating
temperature.

Exposure temperature: The temperature at which the autoclave is held at the desired
operating time. Usually 121 0 C.

Drain temperature: The temperature in the chamber drain that is often used to
control the initiation and completion of various phases of the sterilisation cycle.the
drain temperature should be the coldest point in the entire system, due to the
continual presence of condensate.

Load come up time: The time for the slowest to heat item in the load to reach or
exceed the steriliser set point temperature.

Load cool down time: The time for the slowest to cool item in the load to reach the
desired cooloing temperature.

BOWIE-DICK TEST:

A test to verify that non condensable gases have been removed so that steam can
adequately cover the entire volume. This test will show if the steam has failed to
penetrate due to non condensable gases even if air removal has been achieved.This
test shows whether or not steam penetration is even and rapid and thus by
implication that air or other non condensable gases.it does not confirm that the
sterilisation conditions in the load have been achieved.

UTILITIES:

STEAM:

Plant Steam: General purpose, industrial, non-critical (non-product contact) steam is

generated, distributed from a central boiler. Volatile and / or non volatile additives
added.
Process Steam:
Steam generated using controlled process water to which no volatile / non volatile
additives have been added.
Clean steam:
Steam whose condensate complies with the compendial monograph for Water for
injection
Pure steam:
Clean steam produced from WFI feed water

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Clean steam recommended for ordinary autoclaves .Pure steam is recommended for
all autoclaves associated with parenteral formulations and other sterile products.

Quality of steam should comply with respective site specifications.

PROCESSED COMPRESSED AIR:

Air is a critical utility must be oil free and should pass through a sterilising grade filter
immediately prior to the entrance to the autoclave chamber.

Quality of air should comply with respective site specification

CHANGE CONTROL:

To ensure that once a system, piece of equipment or associated computer system is

validated , that all altera tions or repairs are promptly communicated to Quality
Assurance in order to determine if re-qualification and/or re -validation is necessary

CALIBRATION:

All measuring components used routinely, during qualification and validation need
regular calibration. Calibrating in the operating range of the process gives better
accuracy. They should be calibrated and certified through an accredited laboratory
against refernce standards.The accredited laboratory sends its reference standards
to the national or international institutes for calibration to national or international
standards.These laboratories should furnish methods used for calibration.
Recalibration should be done after any maintenance of these measuring
components.

PREVENTIVE MAINTENANCE:

Sterilis ers are prone to wear and tear, its components not in good condition can
cause an abberant cycle. A log should be maintained which provides a chronological
record of all maintenance activities performed, preventive as well as corrective
Examples of concerns to be addressed in preventive maintenance program
§ Gaskets are properly rated and in good condition
§ Instrumentation is in good operating condition
§ Valves are operational
§ Probes, e.g., level probes, are in good condition and are in the proper location
§ Alarms are functional
§ Insulation is in good condition
§ All safety related features are fully functional

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Typical Preventive maintenance plan:

Prior to routine weekly / quarterly or annual checks : Carry out safety checks
Daily checks
1. Verification of calibration status
2. Leak test
3. Use of chemical indicators
Weekly checks
1. Bowie dick test
Quarterly checks
1. Steam trap
2. Balance thermostatic trap
3. Oil level ( FRL)
4. Magnetic float valve
5. Boiler for any scaling
6. Pneumatic valves
7. Solenoid valves
8. Pressure Switches
9. Electric heaters
10. Overload relays
Annual checks
1. Verification of calibration of
steriliser instruments
2. Chamber overheat cut out test
3. Air filter integrity test
4. As an outcome of performance
qualification

2.0 ABBREVIATIONS:---

3.0 REFERENCES:
PDA Technical report No. 1
HTM 2010

ANNEXURES:

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