1. Autoclaves
Nadina Wiórkiewicz (2011), Small benchtop laboratory autoclave [photograph]. Retrieved from

2. Learning Objectives What materials can be autoclaved?
What Personal Protective Equipment (PPE) is necessary for autoclave use?
How does one properly load the autoclave?
What steps assist in efficient decontamination of biohazardous waste?
How does one verify autoclave performance for decontamination of biohazardous waste?
Skeet, M., & Fear, D. (1995). Care and safe use of hospital equipment (p. 90). London: VSO.

3. 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
Note: burning is the most common form of biowaste disposal in the developing world

4. What can be autoclaved? Surgical Instruments Glassware
Plastic tubes and pipette tips
Solutions and water
Animal food and bedding
Waste
آرمین (2012), Surgical instruments [photograph]. Retrieved from
Pocar19 (012), Gilson Pipette Tip [photograph]. Retrieved from
Lilly_M (2011), Glass beakers [photographs]. Retrieved from

5. Types of waste Waste Type Container What You Need To Do
Biological Infectious Waste
Orange or clear autoclave bag
Autoclave
Medical Waste
Biological Infectious Waste which cannot be autoclaved
Red incineration bag in red waste container
Medical Center- close and place by trash in lab for pick-up, Outside Medical Center- contact Environmental Management for pick-up
Sharps- All needles, syringes, scalpels, blades, and all other sharp medical waste
Sharps container
Animal Carcasses- no biological, chemical, or radiological hazard
Opaque bag
Return to appropriate DLAR location
Animal Carcasses- intentionally infected
Red incineration bag
Incinerate (Medical Center or call Environmental Management pick-up)
Used Biological Culture Media
Glass or plastic
Disinfect with bleach or autoclave

6. Principles of Autoclave Operation
Steam penetrates objects in the autoclave
121 °C (249°F)
Condensation creates negative pressure and draws in additional steam
Moist heat kills microorganisms via coagulation of proteins
Two types of autoclaves
Gravity Displacement
Vacuum Assisted

7. Gravity Displacement
Snelling, Guy. “Autoclave Validation-What is Really Required?.” Retrieved from:
autoclaves/background/Autoclave_validation_-_what_is_really_required.pdf

8. Autoclave Use Basics Personal Protective Equipment (PPE) Packaging
Loading
Operating
User logs
Maintenance logs
Unloading
Improper Autoclave Practices
Nadina Wiórkiewicz (2011), Small benchtop laboratory autoclave [photograph]. Retrieved from

9. PPE for Autoclave Users
Eye Protection
Lab Coat, Buttoned
Closed-toed Shoes
Heat-resistant Gloves
Lilly_M (2011) Laboratory protection goggles [photograph]. Retrieved from
Public Domain Pictures (2014), Retrieved from Pixabay

10. Packaging for Autoclaving
Be sure the material should be autoclaved
No volatile chemicals, sharps, red bag waste, radioactivity
Utilize containers and autoclave bags appropriate for autoclaving
Clear or orange bags
UK Stores   
Do not overfill containers
Tom Beatty (2012), Autoclaves [photograph]. Retrieved from

11. Packaging for Autoclaving
Carefully add water to autoclave bags
Add approximately 1 liter of water to 24” x 30” bag
Aids in reaching appropriate temperature quickly
Creates steam in the autoclave bag to replace cold air
Prepare packaging to allow steam penetration
Check that all containers including bags are vented
Loosely close autoclave bags
Vent lids on bottles containing solutions
Place packaged material in a secondary container
Stainless steel tray or autoclavable polypropylene bin

12. Loading the Autoclave
Load material to allow efficient steam penetration
Autoclave clean items and waste separately
Do not allow material to be autoclaved to touch the sides or top of the chamber
Nadina Wiórkiewicz (2011) Laboratory autoclave [photograph]. Retrieved from:

13. Operating the Autoclave
Be sure the autoclave is functioning properly before use
Record information in User Log
Close door properly and securely
Choose the correct conditions for your material
For decontamination:
Temperatures between °C
Total processing time minutes
Exposure time greater than 20 minutes
Pressure minimum 15 PSI
Select liquid cycle or slow exhaust cycle
ONLY use destined water!!!
Clean with vinegar every day

14. Autoclave User Log Complete User Log with Every Use
Maintain User Logs for Two Years
Complete User Log with Every Use

15. Autoclave Maintenance Log
Maintain Maintenance Logs for Five Years
Complete Maintenance Log for Every Repair
Maintain records of Yearly Calibration

16. Unloading the Autoclave
Put on Personal Protective Equipment
Allow the autoclave to completely finish cycle
Pressure gauge must read zero
Verify cycle conditions were met
Open door slightly to allow steam to escape
Allow contents to cool before removal
Carefully remove items
Be especially careful with fluids and plastic bins

17. Improper Autoclave Practices
MFERMION (2014), Biomedical Waste [photograph]. Retrieved from

18. Common Problems
Build up of a scale due to the use of non-distilled water
Scale may cause the air release valve and the over-pressure relief valve to be clogged
Soaking them for several days in pure distilled water should dissolve any deposits
Hissing noise due a leak in the pressure vessel or valve
Leaky door seal
Thermostat

19. Autoclave Verification Program
Program will ensure that all potentially hazardous biological waste is properly decontaminated prior to deposition in landfill
Program requires testing of the ability of campus autoclaves to kill microorganisms
There are over 50 autoclaves on campus which require this verification program

20. Autoclave Verification
Each Load of Biohazardous Waste
Autoclaves with automatic documentation
Review the printed report
If conditions were met, initial, date, and place User Log number on printed report
If conditions were not met, do not remove the load, repeat autoclave cycle.
If conditions are not met again, autoclave must be labeled out of order.
Contact responsible individual posted on autoclave.
Responsible individual contacts service company to schedule repairs.

21. Autoclave Verification
Each Load of Biohazardous Waste
Autoclaves without automatic documentation
A Chemical Integrator Test Pack (CITP) must be incorporated in each load.
Based on color change the performance of the autoclave will be evaluated.
Tape the autoclaved CITP strip in the appropriate section of the User Log for your load.

22. Autoclave Verification
Monthly Verification for Biological Waste Autoclaves
Biological Indicator Test
Checks all conditions of autoclave cycle, time, temperature, and pressure
Verifies ability of autoclave to kill a microorganism
Utilize the 3M Attest Biological Indicator Monitoring Kit
Record results in Biological Indicator Results Log

23. Biological Indicator Results Log
Maintain Results Logs for Five Years
Complete Results Log with Every Monthly Test

24. Chemical Indicators
These indicators change color after being exposed to specific temperatures.
There are two types of chemical indicators, temperature sensitive tape (autoclave tape) and integrated chemical indicator strips.
Autoclave tape can only attest to the temperature attained and not to exposure time and hence success of sterilization. Integrated chemical indicator strips however, provide a limited validation of temperature and time, though this is not as accurate as a Biological Indicator, and so it cannot be used as a replacement.

25. Biological Indicator Test Procedure
Label Attest package containing biological indicator vial with date, autoclave ID, and load number from user log
Place the indicator vial package in the autoclave with the biohazard waste load
Run the autoclave cycle as usual
Using proper unloading procedures, remove the indicator package from the autoclave

26. Biological Indicator Test Procedure
Label the indicator vial with all information from package
Check indicator vial label for color change from rose to brown
Place the biological indicator vial into the 56°C incubator for hours

27. Biological Indicator Test Procedure
Place a non-autoclaved, labeled control tube in the incubator
Examine the indicator tube at 8, 12, 24, and 48 hours for any color change
Yellow color change indicates bacterial growth
Compare to non-autoclaved control at each time point
Record 48 hour test results in Results Log

28. Learning Objectives Review
What materials can be autoclaved?
Surgical Instruments
Glassware
Plastic tubes and pipette tips
Solutions and water
Animal food and bedding
Waste

29. Learning Objectives Review
What Personal Protective Equipment (PPE) is necessary for autoclave use?
Lilly_M (2011) Laboratory protection goggles [photograph]. Retrieved from
Public Domain Pictures (2014), Retrieved from Pixabay

30. Learning Objectives Review
How does one properly load the autoclave?
Load material to allow efficient steam penetration
Autoclave clean items and waste separately
Do not allow material to be autoclaved to touch the sides or top of the chamber

31. Learning Objectives Review
What steps assist in efficient decontamination of biohazardous waste?
Add approximately 1 liter of water to 24” x 30” bag
Loosely close autoclave bags
Place packaged material in an approved 2° container

32. Learning Objectives Review
How does one verify autoclave performance for decontamination of biohazardous waste?
Each Load
Verify cycle conditions via autoclave printout
CITP indicator use
Monthly
Utilize Attest Biological Indicator Vial