SOP FOR CLEANING AND OPERATION OF ATOCLAVE
A. Associated Risks
Autoclaves, also known as steam sterilizers, operate at high temperatures and
pressures and as a consequence pose a serious risk to operator safety.
Specific risks include:
o Heat, or burns from hot materials and autoclave chamber walls and door.
o Steam burns from residual steam venting from autoclave and materials on
completion of the cycle.
o Hot fluid scalds from boiling liquids and spillage in autoclave
Risk Control/Management
✓ Autoclaves to be regularly inspected and an inspection service and repair
record maintained.
✓ An authorized training session must be successfully completed prior to use of
autoclaves.
✓ Procedure and instruction documentation must be followed
✓ Personal protection must be worn
✓ Heat-insulating gloves that provide complete coverage of hands and forearms
must be
✓ worn when loading and unloading the autoclave.
✓ Eye protection, a lab coat and closed-toed footwear are required to protect
against
✓ steam and scald burns.
OPERATION INSTRUCTIONS
I. Training
All operators must have successfully completed an authorized training
session.
II. Material Preparation
o Ensure that the material is autoclavable. Oils, waxes, some plastics,
flammable materials, and samples containing solvents or substances that
may emit toxic fumes should not be autoclaved.
o Never sterilize items containing chlorides or producing chloride gases in
the automatic autoclaves
o Package material suitably; loosen all lids.
o Bottles with liquids must be a maximum of one-half full and lids
loosened.
o Place items in secondary containers, e.g. Nalgene trays provide in the
autoclave room.
o If your lab has trays confirm with the Departmental Laboratory
Supervisor that they are the correct material.
o Contaminated materials must be secured in a containment vessel or an
autoclavable bag and
3. Log Book
a. Entries must be placed in the log book each time the autoclave is used.
These records are usedfor maintenance/service schedules and reporting
incidents, accidents, and/or faults.
b. Entries should include:
i. Full name
ii. Lab number
iii Date + time
iv Cycle information (gravity/liquid + durations)
4 Loading the Autoclave
✓ Wear heat insulating gloves, eye protection, lab coat, and closed-toed shoes
✓ Place items in secondary container into the autoclave chamber
✓ Ensure there is enough room in the autoclave for steam to circulate. The
bottom rack must always be used to allow steam to circulate underneath the
secondary container.
✓ Close the door firmly.
✓ Close the waste valve (see diagram on white board for valve locations)
✓ Open the steam valve
✓ Set a timer on watch
✓ Allow for 20 minutes for pre-sterilization to ensure proper pressure and
temperatures are reached
✓ Add additional sterilization time based on requirement.
✓ A total of 60 minutes for autoclaving biohazardous waste (infectious and/or
genetically modified materials)
5 Unloading the autoclave
a. Gravity or Fast Exhaust
This method is for solid materials (pipette tips, empty glassware, etc)
Close the steam valve
Open waste valve and allow the chamber pressure to come back to zero
Wearing personal protection equipment, carefully crack door open to release
residual steam
Allow sterilized materials to stand for 1-2 minutes. This will allow steam to clear
and trapped air to escape.
Take out items.
✓ b. Liquid or Slow Exhaust
✓ . This method is for liquid materials or anything that will become liquid while
✓ autoclaving (media, broth, etc.)
✓ . Close the seam valve
✓ . Allow the chamber pressure to come back to zero while leaving the waste
valve closed. This will take approximately 40 minutes.
✓ . Wearing personal protection equipment, open the waste valve and carefully
✓ crack door open to release residual steam.
✓ . Allow sterilized materials to stand for 1-2 minutes. This will allow steam to
clear
✓ and trapped air to escape.
✓ . Take out items.
Waste Disposal and Cleanup
✓ If container says “Biohazard” or is a “Sharps Box” it must be placed in a
Biohazrd collection box after cooling.
✓ All other wastes must be double bagged in a regular garbage pages
✓ Haul trash to a dumpster outside loading dock
✓ Do not dump liquid contain melted agar down the drawn. Instead, collect it in
a disposable container.
✓ Clean up and spills in the autoclave or on the floor.
✓ . Rinse out secondary containers and invert to dry.
✓ Spills inside the autoclave, must be cleaned immediately .
✓ . Report any spills to the Departmental Laboratory Supervisor
Compiled by:Dr Sadhna Pandey, Prof. Of BOT,Govt. KRG PG AUTO college,Gwl, MP, INDIA
