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Autoclave TrainingNovember 16, 2005University of KentuckyDepartment of Biosafety
Learning ObjectivesWhat 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?
Importance of Proper Autoclave UseTremendous 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
What can be autoclaved
Surgical InstrumentsGlasswarePlastic tubes and pipette tipsSolutions and waterAnimal food and beddingWaste
Types of waste
Waste TypeContainerWhat You Need To DoBiological Infectious WasteOrange or clear autoclave bagAutoclaveMedical WasteBiological Infectious Waste which cannot be autoclavedRed incineration bag in red waste containerMedical Center- close and place by trash in lab for pick-up, Outside Medical Center- contact Environmental Management for pick-upSharps- All needles, syringes, scalpels, blades, and all other sharp medical wasteSharps containerMedical Center- close and place by trash in lab for pick-up, Outside Medical Center- contact Environmental Management for pick-upAnimal Carcasses- no biological, chemical, or radiological hazardOpaque bagReturn to appropriate DLAR locationAnimal Carcasses- intentionally infectedRed incineration bagIncinerate (Medical Center or call Environmental Management pick-up)Used Biological Culture MediaGlass or plasticDisinfect with bleach or autoclave
Steam penetrates objects in the autoclaveCondensation creates negative pressure and draws in additional steamMoist heat kills microorganisms via coagulation of proteinsTwo types of autoclavesGravity DisplacementVacuum AssistedPrinciples of Autoclave Operation
Gravity Displacement
Vacuum Assisted
Autoclave Use Basics
Personal Protective Equipment (PPE)PackagingLoadingOperatingUser logsMaintenance logsUnloadingImproper Autoclave Practices
PPE for Autoclave UsersEye Protection
Lab Coat, Buttoned
Closed-toed Shoes
Heat-resistant Gloves
Packaging for AutoclavingBe sure the material should be autoclavedNo volatile chemicals, sharps, red bag waste, radioactivityUtilize containers and autoclave bags appropriate for autoclavingClear or orange bagsUK Stores 6532-0046 Do not overfill containers
Carefully add water to autoclave bagsAdd approximately 1 liter of water to 24 x 30 bagAids in reaching appropriate temperature quicklyCreates steam in the autoclave bag to replace cold air Prepare packaging to allow steam penetrationCheck that all containers including bags are ventedLoosely close autoclave bagsVent lids on bottles containing solutionsPlace packaged material in a secondary containerStainless steel tray or autoclavable polypropylene binPackaging for Autoclaving
Loading the AutoclaveLoad material to allow efficient steam penetrationAutoclave clean items and waste separatelyDo not allow material to be autoclaved to touch the sides or top of the chamber
Operating the AutoclaveBe sure the autoclave is functioning properly before useRecord information in User LogClose door properly and securelyChoose the correct conditions for your materialFor decontamination:Temperatures between 121-124C Total processing time 60-120 minutes Exposure time greater than 20 minutesPressure minimum 15 PSISelect liquid cycle or slow exhaust cycle
Autoclave User Log Maintain User Logs for Two Years
Complete User Log with Every Use
Autoclave Maintenance Log Maintain Maintenance Logs for Five Years Complete Maintenance Log for Every Repair Maintain records of Yearly Calibration
Unloading the AutoclavePut on Personal Protective EquipmentAllow the autoclave to completely finish cyclePressure gauge must read zeroVerify cycle conditions were met Open door slightly to allow steam to escapeAllow contents to cool before removalCarefully remove itemsBe especially careful with fluids and plastic bins
Improper Autoclave Practices
Results of Improper Autoclave Use
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 programAutoclave Verification Program
Autoclave Verification Each Load of Biohazardous WasteAutoclaves with automatic documentationReview the printed reportIf conditions were met, initial, date, and place User Log number on printed reportIf 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.
Each Load of Biohazardous WasteAutoclaves without automatic documentationA 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.Autoclave Verification
Monthly Verification for Biological Waste AutoclavesBiological Indicator TestChecks all conditions of autoclave cycle, time, temperature, and pressureVerifies ability of autoclave to kill a microorganismUtilize the 3M Attest Biological Indicator Monitoring KitRecord results in Biological Indicator Results LogAutoclave Verification
Biological Indicator Results Log Maintain Results Logs for Five Years
Complete Results Log with Every Monthly Test
Biological Indicator Test ProcedureLabel Attest package containing biological indicator vial with date, autoclave ID, and load number from user logPlace the indicator vial package in the autoclave with the biohazard waste load Run the autoclave cycle as usualUsing proper unloading procedures, remove the indicator package from the autoclave
Biological Indicator Test ProcedureLabel the indicator vial with all information from packageCheck indicator vial label for color change from rose to brownPlace the biological indicator vial into the 56C incubator
Place a non-autoclaved, labeled control tube in the incubatorExamine the indicator tube at 8, 12, 24, and 48 hours for any color changeYellow color change indicates bacterial growthCompare to non-autoclaved control at each time pointRecord 48 hour test results in Results Log
Biological Indicator Test Procedure
Biological Indicator Test Results
Learning Objectives Review What materials can be autoclaved?
Surgical InstrumentsGlasswarePlastic tubes and pipette tipsSolutions and waterAnimal food and beddingWaste
What Personal Protective Equipment (PPE) is necessary for autoclave use?
Learning Objectives Review
How does one properly load the autoclave?Learning Objectives Review 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
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 containerLearning Objectives Review
How does one verify autoclave performance for decontamination of biohazardous waste?
Each LoadVerify cycle conditions via autoclave printoutCITP indicator useMonthlyUtilize Attest Biological Indicator VialLearning Objectives Review
References and Linkshttp://ehs.uky.edu/biosafety/autoclave.html
http://www.uottawa.ca/services/ehss/autoclaves.htm
http://www.3m.com/
http://www2.umdnj.edu/eohssweb/aiha/technical/labequipment.htm#Autoclave
http://ehs.uky.edu/em/ Environmental Management 323-6280
Questions or Additional InformationBrandy NelsonSr. Biological Safety SpecialistDepartment of Biological Safety252 East Maxwell StreetLexington, KY 40506-0314Phone: 859-257-3753Cell: 859-699-6082bnels3@email.uky.eduhttp://ehs.uky.edu
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