Autoclave Training Guide

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Autoclave Training Guide

• University of Windsor
• Chemical Control Centre Health and Safety
• B37 Essex Hall.
• Telephone [phone] ext 3523
• www.uwindsor.ca

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Autoclaving Training Outline

• Autoclave Overview
• Autoclave Hazards
• What can be Autoclaved
• How to Autoclave
• Cycle selection
• Proper containers
• Proper loading/unloading
• Autoclave Performance Indicators

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Steam Sterilization

• Pressurized device that uses heat, steam and
  pressure to achieve sterilization or
  decontamination

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Autoclave Overview

• Typically operated at 121C 250F, 15psi, for
  15-45 minutes.
• Allows the heating of liquids above boiling
  point.
• Uses moist heat (steam) to increase efficiency of
  sterilization.
• Heat is used to coagulate proteins, which
  destroys microorganisms and any potential
  biohazard.

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Autoclave Hazards

• Tremendous pressure from steam in chamber
  provides explosive potential.
• High temperatures creates potential for burns and
  scalding.
• Potential exposure to hazardous fumes
• Inadequate decontamination allows for the
  potential of biological hazards to contaminate
  personnel and the environment.

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Autoclave Hazards
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What you CAN autoclave

• Biological waste that can be autoclaved
• Microbial stocks and cultures
• Items contaminated with such waste petri dishes,
  eppendorf tips, pipettes, gloves, paper towel
• Autoclaving is also used for
• Items for sterilizations such as glassware
  media, aqueous solutions

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What you CANT Autoclave

• DO NOT Autoclave
• BIOMEDICAL WASTE
• Including human anatomical or blood waste, animal
  anatomical or blood waste, cytotoxic waste, or
  any waste in contact with these waste products
  (including sharps waste).
• RADIOACTIVE WASTE
• HAZARDOUS CHEMICAL WASTE
• This includes anything contaminated with a toxic,
  volatile, corrosive, or mutagenic chemical
• materials containing solvents, volatile,
  chlorinated compounds (HCL, bleach)
• chemicals (such as phenol, trichloroacetic acid,
  ether, chloroform, ethidium bromoide,
  glutaraldehyde.)
• Check MSDS before hand

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What you CANT Autoclave

• DO NOT Autoclave
• SOME PLASTICS
• Good Choices
• 1. borosilicate glass (Pyrex) has very low
  thermo expansion property and therefore
  resistant to breaking due to heating
• 2. polypropylene (PP) and polycarbonate (PC)
  are heat resistant plastics
• 3. stainless steel is a good heat conductor and
  thus facilitates sterilization
• Poor Choices
• 1. polystyrene (PS), polylethylene (PE) and
  high density polyethylene (HDPE) do not resist
  heat well.
• if there is a risk of material melting ensure
  they are placed in a secondary container which
  is resistant to heat.

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Autoclaves used to treat biohazardous waste at
the University of Windsor

• Biology Room 310
• Essex Hall B35

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How to Autoclave

1. Preparing your items for autoclaving
2. Loading the autoclave
3. Choosing the cycle settings
4. Unloading the autoclave
5. Aborting Autoclave

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Autoclaving Preparing your items

• Sign into log book
• Keeps track of autoclave use for maintenance
  records
• Use personal protective equipment!
• Eye protection
• Heat resistant gloves
• Lab coat

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Autoclaving Preparing your items

• Use a primary container
• Container comes into direct contact with the
  contaminated or non-sterilized material or fluid
• Do not fill more than 75 of holding capacity

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Autoclaving Preparing your items

• Use a primary container (contd)
• Must NOT be a tightly sealed container (might
  explode)
• Primary container must permit heat (steam)
  penetration
• polystyrene (PS), polylethylene (PE) and high
  density polyethylene (HDPE) do not resist heat
  well.
• Loosen screw caps or use self venting caps
• Cap open containers with aluminum foil or muslin
• If using plastic waste bags, leave a small
  opening

Loosen screw caps
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Autoclaving Preparing your items

• Use a secondary container
• Used to contain any spills
• The sides of the secondary container must be
  sufficiently high to contain any spill that may
  occur
• Tray MUST be autoclave safe

For liquids add water
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Autoclaving Preparing your items

• Use temperature sensitive tape
• Will indicate that high temperature has been
  achieved
• Will not prove that decontamination or
  sterilization was successful
• Will assist in keeping track of autoclave and non
  autoclaved items

Before
After
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Autoclaving Loading the autoclave

• NEVER autoclave liquid loads using a solid load
  cycle
• liquids will boil over
• Be cautious if autoclave was recently used
• Rack may be hot
• Use heat protective gloves
• Make sure tray is locked on dolly
• Rack may slide out unexpectedly if not locked

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Autoclaving Which cycle to use

• Autoclaves run 3 types of cycle programs
• The type of cycle depends on what is being
  autoclaved

Liquid/Slow exhaust For autoclaving liquids Prevents liquids from boiling over
Solid/Gravity Best for unwrapped solid items (ie glassware)
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Autoclaving Choosing a Cycle

• For Liquids
• 20 mins / litre of liquid, 5 mins per additional
  litre
• For Solids
• Glassware (empty) 15 mins
• Instruments (utensils) 30 mins
• Biohazardous Garbage at least 40 mins per full
  bag
• Use biological test strips to optimize duration

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Autoclaving Proper loading

• Autoclave biohazardous waste separately
• Do not over load primary or secondary containers
• Allow for sufficient steam penetration
• Do not fill more than 75 to allow expansion
  without overflow
• Do not pack or compress contents

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Autoclaving Unloading autoclave

• Use PPE
• Wait for autoclave to state END CYCLE before
  opening door
• When opening, stand away from door opening
• Make sure no one is standing by door opening

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How to Autoclave Unloading autoclave

• Make sure rack is locked on to dolly
• Unmark any biohazard signs that may be seen on
  waste bags
• Dispose of waste in proper bins
• Please put back heat resistant gloves for other
  users
• Keep autoclave doors shut when not in use

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Autoclave Performance Indicators

• How to know if autoclave is functioning
  correctly

Physical - Annual testing by certified technician - Pressure, Temperature, Cycle times, recorded on paper
Chemical - Heat sensitive autoclave tape - Not an indicator of successful sterilization, useful to keep track of autoclaved and unautoclaved items
Biological - Tests ability of autoclave to sterilize effectively - Bacillus stearothermophilus spore strips often used because they are resistant to steam sterilization. - EZ Test (SGM Biotech) (Fisher Sci 29801 074) - 3M Attest Rapid Readout Biological Indicators - Steris Verify Integrator Laminated and EO Integrators
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Autoclave Issues/Concerns?

• Contact
• Your Supervisor
• Chemical Control Centre
• B-37 Essex Hall, x3523
• Chemistry Coordinator
• Beth Kickham, Chemistry Building, x3527
• Team Leader
• Francis Arnaldo , B37C Essex Hall, x3524