Post on 15-Jan-2016
Autoclave Training Guide
York UniversityDepartment of Occupational Health and Safety
Suite C, East Office Building, 4700 Keele St.Telephone : 416-736-5491
www.yorku.ca/dohs/biosafety
Revised: 16.November 2010
Autoclaving Training Outline Autoclave Overview Autoclave Hazards What can be Autoclaved How to Autoclave
Cycle selection Proper containers Proper loading/unloading
Autoclave Performance Indicators
Steam Sterilization Pressurized device that uses heat, steam and pressure to
achieve sterilization or decontamination
Autoclave Overview
Typically operated at 121°C, 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.
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.
Autoclave Hazards
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
What you CAN’T 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
(e.g.) bleach, formalin, glutaraldehyde, ethidium bromide Check MSDS beforehand
Autoclaves used to treat biohazardous waste at York
Lumbers Room 120A
Farquharson 227A
Chemistry Building 447
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
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
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
Autoclaving: Preparing your items Use a primary container (cont’d)
Must NOT be a tightly sealed container (might explode) Primary container must permit heat (steam) penetration
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 capsSmall opening
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
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
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
Autoclaving: Loading the autoclave
Farquharson: Tighten door so that prongs are
fully extended If noise and/or steam escapes,
abort the cycle and tighten the door more
Lumbers Keep the door button pushed
until the ready prompt is on the screen
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)
Solid/Vacuum * Best for wrapped solid items
Note: for Farquharson, use Solid/Gravity for wrapped, solid items
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 30 mins per full bag
Use biological test strips to optimize duration
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
Unautoclaved Autoclaved (regular) Autoclaved (due to being compressed & sealed)
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
Farquharson: turn door knob slowly and open door slightly to allow steam to escape
Lumbers: make sure your hands are not above the top vents
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
How to Autoclave – Aborting Only qualified personnel should attempt to troubleshoot an autoclave
Farquharson Large autoclave: push abort button Small autoclave: need to manually advance through the autoclave cycle
Lumbers Push abort button
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
Autoclave Issues/Concerns?Contact: Your Supervisor
Farquharson and Lumbers: Jane Grant, Biology, Lumbers 130, x22657 Debbie Freele, Biology, Lumbers 115, x22655 Prof. Roger Lew, Biology, Farq 230, x66114
Chemistry Building: Philip Johnston, Chemistry Building, x33119
York Biosafety Officer: Debbie Kolozsvari, DOHS, x44745
Autoclave Training
Completion of in-class autoclave training is included with the Biosafety Training session.
For practical, hands-on training, contact your supervisor.