Flinders University
School of Nursing & Midwifery
Safe Work Procedure:
Autoclave operation & sterilisation
procedures
Hazards &
Safety measures
SOP Number
RA Number
Contact Person
RA Score
Date:
B:hazardous but controlled
Dec 2014
SOP prepared by:
Review Date :
As information or
procedures change
Jane Stephenson Sue Hollis/Lyn Spencer
General:
 Autoclaves are used in the School for the rapid sterilisation of media and equipment used in microbiology,
culturing and for the decontamination of cultures and biological waste materials.
 Autoclaves operate at high temperatures and pressures and all users must be aware of the hazards:
 All pressure vessels and boilers must be regularly (1-2yrs) inspected and certified, and an
inspection, service and repair record maintained;
 All users must be trained* in the safe operation of the autoclaves prior to first use. This will provide
an understanding of the appropriate conditions for load sterilisation, and an appreciation of the
hazards associated with heat, steam and pressure.
 Before any staff member may operate the autoclave a training sessions will be conducted by
competent persons
Hazards Identified
 Burns & scalds from steam, hot fluids &
materials
 Burns from radiant heat and hot surfaces
 Manual handling of containers
 Pressure vessel rupture
 Poor housekeeping – trips
Poor housekeeping – slips from spills/leaks
Rating
M
Safety Measures
 Training; containment baskets; protective
gloves/clothing/footwear; mandatory signage;
 Training; containment baskets; protective
M
gloves/clothing/footwear; mandatory signage;
M
 Use trolleys
 Pressure vessel certification and regular
M
maintenance
M
 Keep aisle clear;
Hazard signage; mop & bucket to clean spills;
safety mat in front of autoclave
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Material Preparation:
Preparation of ‘clean’ materials for Sterilisation:
 Ensure that the material is autoclavable:
 Hazardous chemicals must not be autoclaved;
 Materials that cannot generate their own steam or be penetrated by steam such as oils and waxes
cannot be sterilised by autoclaving;
 Autoclaving is suitable for the sterilisation of heat-stable:
 Materials that can be penetrated by steam, liquids that can generate their own steam and materials
with solid surface eg: filter apparatus, pipette tips;
 Glass: must be borosilicate (‘pyrex’) to withstand the heat and pressure;
 Plastics: polycarbonate (PC) is autoclavable; most polypropylene (PP) is autoclavable but some may
distort and both will deteriorate with repeat autoclaving over time;

Bottles with liquids must be a maximum of ⅔ full, lids loosened or covered with a steampenetrable bung or loose cover eg foil;

Solid goods (instruments, pipette tips) must be wrapped in steam-penetrable paper or
loosely enclosed in aluminium foil or a container with a loose-fitting lid so that steam can still penetrate;

All items must be stabilised in a stainless steel mesh basket with a raised base to
support goods above the floor of the autoclave;
Preparation of Biohazardous/Biological materials for Decontamination:
 Animal carcasses must NOT be autoclaved – they must be disposed of by incineration;

Biologically contaminated materials and waste must be enclosed in an autoclavable
Biohazard bag - the bag must be open to allow the penetration of steam and the displacement of air then placed in a solid stainless steel container to confine spillage (no lid) prior to autoclaving;
 Biologically contaminated materials must be processed as soon as possible;
 NOTE: Spillage of molten agar WILL clog steam outlet valves, preventing completion of cycles and
incurring high repair costs.
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Autoclaving:
 All operators MUST:
 wear protective clothing, closed footwear;
 use heat-insulating/moisture-proof gloves (provided) for use when loading and unloading;
 wear a full-face shield (provided) for use when unloading goods;
 follow this procedure, and individual instruction documentation;
 ensure that flat container surfaces are not in direct contact with the autoclave chamber surface;
 Place pressure-sensitive tape on goods or container;
 enter details of the process and user in the log book;
 enter any faults encountered during the cycle.
Loading:
- Wear heat-insulating gloves;
- place basket into autoclave chamber;
- close door firmly but do not overtighten.
Select cycle:
FLUID cycle: Standard: 15’@121psi;
Increase time for very large volumes
DRY cycle: Standard 20’@121psi / 20’ drying time;
Increase time for plastics
Hard Goods (Biological Decontamination) cycle: Standard 30’@121psi;
Increase time for very dense materials eg: soils
Operating the autoclave:
Biologically contaminated materials and waste must be enclosed in an autoclavable
Biohazard bag
 Autoclave in ‘kill’ cycle (30’@121psi recommended) to eliminate contamination risk and to compact
contents; very dense materials eg: potting soil, use 45’@121psi
After autoclaving:
CLEAN GOODS - place sterile goods on rack to cool
DECONTAMINATED WASTE/GOODS:
 Allow waste to cool prior to storing while awaiting disposal by Sturt caretakers
 NB: Excess liquids can be decanted to the sewer after autoclaving;
 Do not overfill bins; DO NOT place corrosives, hot items or fluids in bins;
 dispose of waste as soon as possible - and clean up any spills;
clean up spills with mop and dispose of any waste appropriately.
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Applicable Standards and References
Standard/
Reference
Title
AS/ANZ 2243.3 6.6
AS 1210
AS 2192
Safety in laboratories - microbiological aspects and containment facilities
Pressure vessels
Sterilisers-steam-downward displacement
SOP Authorisation
Supervisor
Name:
Jane Stephenson
Signature:
Date:
23/12/14
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