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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