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.