Produced by the Health and Safety Department, the University of Edinburgh

Autoclaves
There are several different types of hazards associated with the use of
autoclaves. The main ones are the pressure vessel hazard, the unloading
hazard and failure of a make-safe (cycle to sterilise contaminated waste)
process. The following describes some of the main factors that should be
taken into account in risk assessments and the types of control measures to
be implemented.
The relevant British Standard covering use of autoclaves in laboratories is BS
2646 Autoclaves for Sterilisation in Laboratories 1993. When Schools are
purchasing autoclaves or arranging maintenance work they should always
check the autoclave and the associated installation and servicing complies
with the British Standard specifications. A simple open tun-dish is not suitable
for the exhaust line of a laboratory autoclave. The need for any additional
controls on the exhaust for autoclaves in Containment Level 3 laboratories
should be determined by risk assessment .
Regular maintenance is essential for the continued efficiency and safety of
laboratory autoclaves and ancillary equipment. Schools should have a
continuing programme of planned preventative maintenance throughout the
life of the autoclave. Where the autoclave is used to make safe waste it is also
necessary to validate and monitor the performance of the autoclave. Further
details on this are given in section 8 below.
In addition to ongoing maintenance and validation arrangements, it is a
requirement under the The Pressure Systems Safety Regulation that
autoclaves be periodically examined by a competent person. This
examination is necessary in advance of the autoclave being brought in to
service and thereafter at appropriate regular intervals. Within the University
this is carried out by the University's Engineering Insurers and all autoclaves
must be notified to the University's Engineering Insurance Surveyor in order
that they can be added to the schedule and arrangements be made for
examination. Notification of autoclaves should be made through the University
Health and Safety Department.

1. Operation of Autoclaves
Autoclaves should be operated only by persons who have been trained and
instructed in their use.
Operation instructions should be provided for the operator and should include
details of action to be taken by the operator in the event of a fault or any
abnormality in autoclave performance. It is recommended that simple, easy to
understand instructions are provided in addition to the detailed manual
provided by the manufacturer.

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Produced by the Health and Safety Department, the University of Edinburgh

2. Maintenance
The continuing safe and effective use of the autoclave depends on a
programme of planned maintenance throughout its life. Maintenance
schedules should therefore follow all of the recommendations in the current
British Standard. The manufacturer should always be consulted on
maintenance intervals. Schools should have a continuing programme of
planned preventative maintenance throughout the life of the autoclave.
When a fault occurs during a make-safe cycle an assessment of risk should
be made and appropriate action taken. It may be necessary to disinfect those
chamber attachments on which engineering work is to be carried out. During a
make-safe process, chamber condensate should be considered to be
contaminated with viable micro-organisms.
A contaminated laboratory autoclave should never be returned to the
manufacturer for servicing or repair. Decontamination should be carried out in
accordance with local safety rules. The use of permit to work systems for
maintenance contractors is strongly recommended.

3. Protective Clothing
A protective laboratory coat of side or back fastening style should be worn in
the autoclave loading/unloading area(s).
Additional protective clothing should be available in the loading/unloading
area(s) to protect the operator. This should include an impervious apron, heatresistant gauntlet gloves, suitable heavy-duty footwear or overshoes and a
full-face visor. The hazards on loading include spills of biohazardous material,
broken glass and dropped load contents. The hazards on unloading include
splashes and spillage of hot material from the load, hot condensate, hot
equipment, broken glass, dropped load contents, and vapour from volatile
chemicals.

4. Loading the Autoclave

All materials awaiting autoclaving should be stored safely. Bagged waste
materials should be supported in a robust, leakproof container, bags should
never be placed directly on the floor.
It should be possible to identify the source of all materials to be autoclaved. A
labelling system should be used in order that the waste can be tracked back
to the laboratory or area from where it came. This serves two important
purposes. Firstly to be able to identify the contents of the waste and what
hazards may be present in the event of someone having an accident in which
they are exposed to any infectious agents that may be present. Secondly to
be able to identify the producer of the waste if there is any problem with it,
such as a container is leaking, overfilled, or contains inappropriate items etc,
to prevent repetition of bad practice.

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Items should be packed in a way that ensures that steam will penetrate the
load. Bags should not be sealed, if the tops have been taped or tied the bags
should be opened or slashed to allow effective steam penetration.
When some chemicals (including disinfectants) or materials are autoclaved
they are likely to produce vapours that could harm persons exposed when
opening the door or they may be corrosive to the autoclave. A risk
assessment should be made of all items to be autoclaved taking account of
this possibility.

5. Unloading the Autoclave

Temperature and pressure indicators and warning lights should be checked to
ensure that the autoclave has successfully completed the operating cycle. If a
fault is indicated, attempts to open the autoclave should only be made with
the authority of the responsible person.
It is dangerous to attempt to release the autoclave door mechanism before
the chamber is vented to atmosphere or whilst the load contents are at high
temperature. No attempt should be made by the operator to override door
interlocking safety devices.
The operator should stand clear when opening the door as hot liquid or
vapour may escape from the chamber. The operator should also be aware
that containers of liquid could be pressurised and may explode, volatile liquids
may produce harmful vapour and liquids spilled on unloading may cause
scalding. The following measures are designed to minimise the unloading
hazard:

Temperature activated door interlocks;

Timer activated door interlocks;
Training and supervision of operators;
Protective clothing; and
Load transfer systems.

After autoclaving, waste containers should be emptied in a safe manner and

their contents disposed of or reclaimed as specified by local laboratory rules.

6. Waste Containers
All waste should be placed in waste containers which should be easily
transportable, leakproof and of a robust design with solid sides and bottoms.
They should allow adequate steam penetration to the contents.
If autoclave plastic bags are used they should be supported in a waste
container whilst in the laboratory, during transport to the autoclave and also
whilst in the autoclave. The bag should be open during the autoclave cycle so
that steam can penetrate its contents. It is recommended that transparent
(rather than opaque) autoclave bags be used as this allows the autoclave

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operator to see if any hazardous materials have been placed in the bag by
mistake.

7. Operating Cycles
Typical operating cycle conditions are listed in the table below.
TYPICAL OPERATING CYCLE CONDITIONS

Process

Sterilising
Temperature

Holding Time
(minutes)

Min 0C

Max 0C

Min

Max

Liquids Sterilisation

121
115

124
128

15
30

Equipment/Glassware
Sterilisation

121
126
134

124
129
138

15
10
3

Make-safe (waste)

121
126
134

125
130
138

15
10
3

The operating cycle should take account of heat up times. The time required
for the load to reach sterilizing temperature should be determined during
validation tests.

Operating Cycles for Liquids Sterilisation

When liquids are sterilised the possibility of adverse effects on the liquid
caused by the heat treatment must also be taken into account on selecting a
cycle. The operating cycle should be selected to ensure that sterilisation is
achieved with minimum damage to the liquid. Microbiological culture media
are particularly heat sensitive; the degree of deterioration is related to the
length of time the medium is maintained at sterilising temperature; the heat-up
and cooling stages also contribute significantly to this deterioration.
Heat-up times should be as short as possible, achieved by uniformly filling the
chamber with steam at sterilising temperature. Large volumes of fluids will
heat up slowly, therefore volumes of liquids should be kept small; a maximum
container volume of 500ml is recommended, larger volumes taking
considerably longer to heat up (and cool down).
Cooling loads quickly helps to protect heat-sensitive constituents and also
shortens operating cycle times. Air at high pressure may be admitted to
ballast the chamber, minimise boiling and prevent bottles exploding.
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Containers should be loosely capped unless they are specifically designed for
sealing. However, sealing bottles can increase the likelihood of explosion
during autoclaving and slows cooling.

8. Validation and Routine Monitoring of Autoclave Performance

i) Validation
The purpose of validation is to test the ability of the autoclave effectively to
perform the autoclaving process when loaded with user-defined loads. An
autoclave cycle can be split into three phases, warm-up, holding and cooling
down. Validation should demonstrate that in the holding part of the cycle the
necessary temperature has been attained throughout the load and this is held
for a minimum time. Calibration, which should not be confused with validation,
compares the temperature set on the dial to the temperature in the chamber.
If an autoclave is used for a waste cycle (also commonly called a make safe
or destruct cycle) for loads containing pathogens or harmful genetically
modified micro-organisms then validation is required to demonstrate that the
process is effective in achieving a 100% kill. A worst case load of simulated
waste (ie worst case volumes, materials and equipment but not contaminated
with pathogens) should be placed in the autoclave for the validation test.
Probes are then inserted at various (12) points in the waste and connected to
recording equipment. This then shows the temperature at the various points
throughout the cycle. The holding part of the cycle commences once all
sensors indicate the prescribed sterilizing temperature has been reached.
All sensors must then maintain at least that temperature for the prescribed
holding time.
Validation must be carried out by a competent trained person using
thermometric test equipment that is calibrated and traceable to national
standards. The test method is described in British Standard 2646. The use of
biological or chemical indicators for this purpose is not acceptable. The use of
benchtop autoclaves for waste is not recommended because these cannot
usually be validated.
The prescribed operating cycle conditions for a make safe cycle for waste are:
i) at least 121oC (max 125oC) for 15 minutes; or
ii) at least 126oC (max 130oC) for 10 minutes; or
iii) at least 134oC (max 136oC) for 3 * minutes.
*Loads comprising a variety of items and containers do not heat uniformly.
Short holding times are therefore subject to large proportionate variations and
should be avoided if possible.

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Following validation it will be possible to ascertain the settings required on the

autoclave to achieve the above conditions. This will vary considerably with
many factors including the load characteristics. For example, one particular
autoclave may need to be set at 121oC for 50 minutes in order to achieve
cycle (i) above whereas another may need to be set at 123oC for 75 minutes
to achieve the same. Where dials can be altered by the user, the temperature
and time requirements for a waste cycle should be clearly displayed on the
autoclave.
Validation must be carried out at least annually and at any other times when
the previous test may no longer be valid (such as part of re-commissioning
after maintenance work). The person responsible for the autoclave should
check certificates and print-outs received for the validation testing to satisfy
themselves that the autoclave meets the criteria specified above for the
required cycle(s). Validation records should be kept for 5 years. Health and
Safety
Executive Inspectors are likely to request sight of, or copies of, records during
visits to the University.

ii) Routine monitoring

Following validation, autoclave performance should be routinely monitored.
Where autoclaves are used for waste containing pathogens, the autoclave
should be fitted with chart recorders or print out facilities and these should be
checked after each run and kept as part of the autoclave process record.
Autoclave tape and indicators printed on bags do not show the load has been
sterilised. These are used only to indicate that a load has been processed
since the stripes appears very quickly even at low temperatures. Use of these
would not be regarded as an adequate means of monitoring autoclave
performance.
Biosafety Unit
Health and Safety Department
University of Edinburgh
August 2003
Note: The information contained within these pages is for use by University of
Edinburgh personnel only.
If you wish to discuss any issues relating to the above documents please
contact the University Biological Safety Adviser.

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