Procedures for Using Steam Autoclaves
QUESTIONS: Contact DRS, 217-333-2755, for additional information.
MAINTENANCE & CARE:
Check the Pressure:
Check all pressure gauges.
Jacket pressure gauge
should be a minimum of 15
psi (refer to
manufacturers
instructions for maximum
psi).
Use Sterilization Indicators
Visual indicators, e.g.,
chemical/tape indicators,
measure one or more
physical conditions of the
autoclave cycle.
Mechanical indicators
record the time-
temperature profile
attained during a cycle.
Biological indicators, i.e.,
spore vials, are the only
approved way to prove
sterility.
NOTIFY FACILITIES SUPPORT
WHEN A PROBLEM OCCURS:
No steam.
Valves leaking.
Door gasket deterioration.
Jacket pressure below 15 psi.
Temperature not reaching
121C (250F).
Erratic temperature or
pressure during cycle.
Steam escapes from around
door during cycle.
Water on floor or in chamber
after a cycle.
Articles very wet after a fast
exhaust or dry cycle.
Excessive steam in area
during cycle or after opening
door.
CHOOSE PROPER OPERATING CONDITIONS: DECONTAMINATING
Sterilization will occur only when the conditions of time, temperature, pressure, and humidity have been met. BIOHAZARDOUS WASTE BAGS:
Incorrect selection of time or exhaust cycle may damage the autoclave, cause liquid to boil over, or bottles to Use autoclavable, high
break. strength polymer bags
Proper use of autoclave will minimize the chance of serious injury. imprinted with chemical
indicator (if possible).
Biological Waste Liquids Dry Items Glassware Add approximately 250 mL
Items
(Gravity Cycle) (Liquid Cycle) (Gravity Cycle) (Gravity Cycle) water to bag before closing
and transporting to autoclave
Loosen caps or use
Open the bag >2", Dirty: Place in area.
a vented closure, Fabrics Wrap;
Place in tray, middle of the pan;
Preparation Fill containers no Instruments: Clean, Place bag in a polypropylene
Place indicator if Clean: wash, rinse,
more than 75% dry, lay in pan or stainless tray before
needed wrap
capacity
autoclaving.
Dirty: In detergent
Fabrics: Separated, Open the bag at least 2 to
Placement in and pan;
In the center Upright in pan on edge;
Autoclave
Instruments: Flat
Clean: On side or allow steam to enter.
inverted When complete, the
sterilization indicators must
Temperature 121oC 121oC 121oC 121oC show adequate
decontamination before
60-120 min. 22 min. for volumes disposal as municipal solid
Treatment Time in depending on load <100mL; waste.
30-60 min. 30-60 min.
Minutes size and packing 40 min. for volumes
density >100mL
Repeat cycle if sterilization
indicators do not show
Dirty: Slow exhaust; evidence of sterilization.
Exhaust Cycle Slow exhaust Slow exhaust Fast exhaust and dry
Clean: Fast/dry Overbag biohazardous waste
bag with an opaque trash
Avoid puncturing Hot bottles may Check reference for Glassware with bag before placing in the
Notes: bags. Overbag and explode. Let cool proper packaging cracks or deep
dispose of properly. before moving. methods scratches may crack regular trash.
CAUTION: Reminders
NEVER AUTOCLAVE FLAMMABLE, REACTIVE, CORROSIVE, TOXIC or RADIOACTIVE MATERIALS, e.g., Store biohazardous waste in
bleach. a closed leak-proof
Materials that melt (plastic lab wear) at 121C will block chamber exhaust drain if not placed in a container.
shallow autoclave pan able to withstand that temperature. Never allow waste to
o
Use caution when increasing autoclave temperature to 135 C because plastics (including some plastic accumulate in the lab.
pans) melt at this temperature, causing difficult clean-up and damage to temperature sensors. Never leave waste
Always wear safety glasses, goggles, or face shield, lab coat or apron, and heat-protective non-asbestos unattended.
gloves when opening door or removing item(s) from autoclave. Clean up leaks and spills with
Reference:
a suitableUsing the Gravity
disinfectant.
Do not mix loads that require different exposure times and exhaust.
Displacement Steam Autoclave in
Open door only after chamber pressure returns to zero. Leave door open for several minutes to allow
the Biomedical Laboratory
pressure to equalize and for materials to cool. DHHS/PHS/HIH/DS
Open door slowly. Beware of rush of steam or water.