autoclave training
TRANSCRIPT
Fundamental Autoclave
Techniques
Version 3, September 2010
Veronika Tatarinoff
GSBmE University of NSW
Brandy Nelson
University of Kentucky
The following information is
directed at aseptic/sterile
surgical, tissue culture
(OGTR/PC2)
and QC1 techniques at the
GSBME.
GSBME Autoclave Training Program
This presentation forms part of the GSBME
Autoclave training program.
1. Fundamental Autoclave Techniques (FAT)
2. Review of supporting literature
(SOP, SWP, RA)
3. FAT quiz
4. Demonstration of GSBME Autoclave use
5. Competency assessment
6. Authorisation sign off in WITA
Learning outcomes :
At the end of this session participants will be able to: Describe the mechanisms of autoclave sterilisation
Identify autoclave/s at GSBME
Identify materials that can be autoclaved
Package and label items for sterilization
Correctly load/unload the autoclave
Identify and select appropriate sterilisation cycle/s
Verify autoclave sterilisation process
Maintain log of sterilisation
Dispose of biowaste appropriately
Identify and use the Personal Protective Equipment (PPE) and devices in a safe manner.
Identify hazards and risks to self and others, improper autoclave techniques and identify appropriate responses to injury.
What does ‘aseptic’ mean?
“preventing putrefaction [rotting, decomposition]
by securing the absence of bacteria” Oxford Dictionary
Disinfection.
The process of destroying
or inhibiting the growth
of micro-organisms on a
surface.
Sterilisation.
The destruction of all living
micro-organisms,
including viruses and
spores. This is an
absolute state.
Micro-organisms
Include bacteria, viruses, fungi & spores
Found on every external surface including respiratory tract and alimentary canal
A micro-organism “infection” occurs when the micro-organism grows in an area where it isn’t normally found or in numbers greater than normal.
Surgical and cell culture procedures provide an ideal opportunity for micro-organisms to cause problems.
Sterilisation methods
There are a number of methods which can kill
or remove micro-organisms including:
Heat (dry or steam – autoclave)
Gas (eg Ethylene Oxide)
UV or gamma irradiation
H202 plasma
Liquid sterilisation (eg glutaraldehyde)
Filter sterilisation (liquids only)
Fundamental autoclave information
Personal Protective Equipment (PPE)
Packaging
Loading / Unloading
Operating cycles
User logs
Hazards
Improper Autoclave Practices
Autoclave use basics
Critical Sterilisation parameters
Autoclaving uses high temperature, high
pressure and steam to kill organisms.
The critical autoclaving parameters are:
temperature
steam under pressure
time
Items have to be hot enough for long enough
in the presence of steam to disrupt the
proteins of the micro-organisms.
Water is boiled under pressure and the steam is
enclosed within the chamber.
Objects placed in the chamber and the steam
circulates around and penetrates them.
Temperatures of 121-134°C at set pressure for
20-60 minutes are generally used.
Condensation creates negative pressure
and draws in additional steam.
Moist heat kills microorganisms
via coagulation of proteins.
Autoclave (steam) sterilisation
What can be autoclaved?
Surgical Instruments
Glassware
Plastic tubes and pipette tips
Solutions and water
Animal food and bedding
Waste
What can’t be autoclaved?
The effectiveness of autoclaving depends on steam, the steam must be able to penetrate the material being autoclaved. Goods that are packed in impervious packaging cannot be autoclaved.
Because damp heat is used, heat labile products (such as some plastics) cannot be sterilised in this way or they will melt. Items that may be damaged by steam, should be sterilised by another method.
Chemicals (e.g. bleach) emit toxic fumes when autoclaved
Incompatible chemicals e.g. acids , bases , organics solvents
It is essential to ensure that your material/s can be autoclaved.
PPE for Autoclave Users
Eye Protection
Lab Coat, Buttoned
Closed-toed Shoes
Heat-resistant Gloves
Packaging for Autoclaving
Be sure the material can be autoclaved
No volatile chemicals, acids, bases, sharps
Utilize containers and autoclave bags appropriate for autoclaving
Clear or yellow “biohazard“ bags
Autoclave paper bags/roll
Metal bins or containers
Pyrex or Shott glassware
Plasticware
Do not overfill containers
Do not tighten lids
Label – Identify contents, owner & date
Loading the Autoclave
Load material to allow
efficient steam penetration
Autoclave clean items and
waste separately
Do not allow material being
autoclaved to touch the
sides or top of the chamber
Ensure load probe cord is
not in trolley path to prevent
damage
GSBME AUTOCLAVE USE
Consult with the Animal House manager if animal surgery is in progress before using the autoclave.
Surgery has priority.
ALWAYS CHECK Sterilisation parameter SETTINGS BEFORE USE.
Different materials or quantities may require different sterilisation temperatures and times. There is a guide on the wall opposite the autoclave and in SOP S006.
Specific instruction on operation of each GSBME autoclave is given in SWPI005 & I006 and will be demonstrated to you.
GSBME Autoclaves
Getinge HS55 (automated) Atherton Benchtop model
(small manual)
Getinge HS 55 Automated
This Autoclave is a large stand alone model.
It is controlled by a microprocessor and has both set programs and manual parameter selection.
If your material requires specific parameters please see autoclave custodian.
P02 - POROUS CYCLE
USE: DRAPES / WRAPPED PACKAGES / BOTTLES
Items that are required to be dry at the completion of
the cycle and/or requiring greater steam penetration.
PARAMETER SETTINGS
STERILISE 121C
16 minutes
DRYING TIME 5 minutes
DRYING AIR PULSE 10 minutes
P09 - HARD GOODS CYCLE
USE: INSTRUMENTS/SOLID ITEMS
Items of equipment that do not require
wrapping, drying or prolonged cooling.
PARAMETER SETTINGS
STERILISE 121C
16 minutes
DRYING TIME 5 minutes
P04– FLASH (HARD GOODS)
CYCLE
USE: INSTRUMENTS/SOLID ITEMS
Items of equipment for surgical procedures
that do not require wrapping, drying or
prolonged cooling .
PARAMETER SETTINGS
STERILISE 126C
12 minutes
DRYING TIME 3 minutes
Liquids
Liquids take longer to heat up and cool down than
most solid materials.
A control (similar fluid) at the maximum volume of
sample load is required for autoclaving liquids. The
load probe is placed in the control bottle to ensure
that the liquids reach the correct temperature for the
correct time to ensure sterilisation.
Lids must be loose to prevent explosion.
Liquids must be in heat treated laboratory glassware
with room for fluid expansion.
A liquid cycle must be used to prevent boiling over.
LIQUID CYCLE
P05 – Fo Liquids, open USE: SALINE, WATER
Non heat sensitive liquids that require a probe in a
control volume to ensure specified parameters are
reached. A slow pressure release is used to avoid
boiling over and allow cool down
PARAMETER SETTINGS
STERILISE 121C
16 minutes
DRYING RAMP TIME 0.05/m
Ensure that lid is loose on the bottle/s during
sterilisation
P10 - WASTE CYCLE
USE: OGTR/PC2 TISSUE CULTURE WASTE BINS
Items in the bins are contaminated with biological materials and do not require wrapping, drying or prolonged cooling.
PARAMETER SETTINGS
STERILISE 121C
60 minutes
DRYING TIME 5 minutes
Following completion & verification of sterilisation, waste must be double bagged, labeled and placed into yellow bins in 4°C cold room pending disposal by UNSW contractor.
Bench top Autoclave
The GSBME small benchtop model is a totally
manual, gravity displacement style of
autoclave with no drying or cool down cycle.
Modification of parameter settings is made by
adjusting gauges directly on the autoclave.
Primarily this autoclave is a back-up if
required.
Autoclave User Log
User Log must be completed with every use
User Logs are maintained for at least five years
Date Time Authorised User Cycle Type e.g. fluid,
hard goods
Items /comments eg drapes,
bottles, waste, validation
Unloading the Autoclave
Put on Personal Protective Equipment
Verify cycle conditions were met
Stand clear of door when opening
Allow contents to cool before removal
Carefully remove items
Be especially careful with fluids and plastic
bins
INDICATORS can be used to ensure an autoclave has reached the correct temperature and pressure for the correct amount of time.
They fall into 3 broad categories:
* Chemical * Biological * Physical
How can you tell if something
has been sterilised?
Indicators
Chemical indicators, can be found on medical packaging and autoclave tape, and these change colour once the correct temperature conditions have been met. This indicates that the object inside the package, or under the tape, has been exposed to heat.
Physical indicators often consist of an alloy designed to melt only after being subjected to 121°C for 15 minutes. If the metal has melted, the change will be visible.
Biological indicators include Attest devices. These contain spores of a heat resistant bacterium. If the autoclave did not reach the right temperature, the spores will germinate, and their metabolism will change the colour of a pH-sensitive chemical.
Printout & gauges. As well as these separate indicators, autoclaves have temperature and pressure gauges visible from the outside. A printout maybe available to confirm conditions were met.
Example of a Chemical indicator
Autoclave tape - colour change(bottom) indicates a
temp of 80°C has been reached.
Examples of Physical indicators
Untreated physical indicators
Class 5 & 6
Colour change in treated
(bottom) indicator,
sterilisation confirmed
Examples of Biological
indicators
Unprocessed Processed - Failed Processed - Passed
Autoclave Cycle Verification
Each Autoclave Load (automatic documentation)
Have the sterlisation conditions been met?
Document all autoclave runs in the User Log
Review the printout report & any other indicator that
you may have used.
If conditions were met, remove the load. Printout
report can be filed in lab book.
For validation cycles indicator and print our are filed in
the equipment validation log.
If conditions were not met, do not remove the load,
review and repeat cycle.
If conditions are not met again, contact
Autoclave custodian.
Tie waste bag with string, leave bag in metal biohazard bin
Fix autoclave tape to bag
Leave bin lid off to one side for autoclaving
It is essential that each biowaste load has been verified as sterile prior to transfer to cold room pending removal by UNSW waste contractor.
Indicator in the centre of the load
Packaging OGTR/PC 2 Waste for
Autoclaving
OGTR/PC2 Biowaste Bins
Over full bin
Hazards associated with using
autoclaves
Burns from hot autoclave, sterilised items, steam.
Sharp/hot/heavy items falling trolley onto operator.
Cuts from broken glass
Possible noxious/toxic fumes released from melted
plastics
Biohazard contamination (OGTR/PC2 waste)
Chemical incompatibility
In an emergency call x56666 security.
Importance of Proper
Autoclave Use
Tremendous pressure from steam in chamber provides explosive potential
High temperatures and presence of extremely hot water creates potential for burns and scalding
Inadequate decontamination allows for the potential of biological hazards and personnel and environmental contamination
Autoclave
Explosion
Results of Improper Autoclave Use
Improper Autoclave
Practices
Additional Information
It is essential that you read and understand the following GSBME documents prior to your FAT quiz.
1. SOP S006 - Guideline to sterilisation at GSBME
2. SWPI005- Operation of getinge autoclave SWPI006- Operation of small autoclave
3. Risk Assessment - Basic use of Autoclaves
It is essential that you have completed PC2 training, quiz and authorisation prior to entry to PC labs at GSBME.
It is essential that you have completed UNSW Animal Care and Ethics training and have ACE approval to conduct any animal work.
Where do I go from here?
This “fundamental autoclave techniques” is followed
by a quiz. Download from OHS page on GSBME
School Website:
http://www.gsbme.unsw.edu.au/ohs/autoclave
Submit your completed quiz to either the Lab or
Animal Research Facility Manager who will arrange:
Demonstration of autoclave/s use
You will then complete a competency assessment
Authorisation for autoclave use occurs when “Signed
off” as competent by the trainer.
Don’t forget your PPE!