Instructor Vinod Jyothikumar,PhD Director-Office of Lab Safety [BSO, RSO, LSO] Email: [email protected] Tel # 202-994-8258 “A safe, healthful, and secure environment for scholarship and research.” Principles of Autoclave Operation •Steam penetrates objects in the autoclave •Condensation creates negative pressure and draws in additional steam •Moist heat kills microorganisms via coagulation of proteins Items that can be Autoclaved • Biohazardous waste • Media and other non flammable liquids • Instruments • Laboratory Glassware • Tubes and pipette tips • All autoclaved items must be placed in a secondary tray inside the autoclave Potential Hazards of Autoclave Use • Burns - high temperatures and very hot water/steam presents the potential for burns and scalding • Incomplete Decontamination - Improper use and/or autoclave malfunction can cause incomplete sterilization, leading to contamination with biological hazards. • Pressurizing & rupture of closed vessels inside autoclave or on opening of door. Closed vessels can explode Potential Hazards of Autoclave Use Opening Door Too Quickly • Can cause even Pyrex containers to crack (spilling/splashing very hot liquid) • Can cause rapid violent boiling of liquids because they are higher than boiling point now. • If load became unstable, items might fall over or fall out and splash/spill. • If inner container has been pressurized (inadvertently) it may burst. Preventing Hazards with Autoclaves Use Proper PPE • Eye protection • Lab Coat • Closed toe shoes • Heat Resistant Gloves • Use a protective apron and face shield if removing liquids from the liquid cycle Preventing Hazards with Autoclaves • Read, and understand, the owners manual as manufacturer recommendations vary. • Make sure autoclave doors and gaskets are firmly locked into place before operating the autoclave. • These interlocking mechanisms help to prevent a sudden release of high pressure steam. Preventing Hazards with Autoclaves • Place a warning sign saying, “caution hot surfaces” in the vicinity of the autoclave. • Never store combustible materials near the autoclave. • Never autoclave toxic, volatile, or radioactive materials. • Remove solutions from the autoclave slowly and gently. • Some solutions have the potential to boil over when moved or exposed to room temperature. • Clean up spills immediately. • Report malfunctions or accidents to your supervisor immediately. Autoclaves in SEH Building Steris – Amsco Century Medium Steam Sterilizers Cycles 1. Prevacuum Cycle 2. Gravity Cycle 3. Liquid Cycle Autoclaves in SEH Building Steris – Amsco Century Medium Steam Sterilizers Touch Screen Autoclaves in SEH Building Steris – Amsco Century Medium Steam Sterilizers Touch Screen Autoclaves in SEH Building Steris – Amsco Century Medium Steam Sterilizers Prevacuum Cycle Recommended for: • Heat and moisture stable goods • Decontamination of biohazardous waste Autoclaves in SEH Building Steris – Amsco Century Medium Steam Sterilizers Gravity Cycle Recommended for: • Glassware 15 min. • Instruments – with any porous material 20 min. • Hard goods • Wrapped 15 min • Unwrapped 30 min Autoclaves in SEH Building Steris – Amsco Century Medium Steam Sterilizers Liquid Cycle Recommended for: • Non-flammable liquids • Must be in vented containers • Borosilicate glass • Metal 121 °C for 45min. Autoclaves in SEH Building Steris – Amsco Century Medium Steam Sterilizers Autoclaves in SEH Building Steris – Amsco Century Medium Steam Sterilizers End of Cycle • Tone sounds • Display Shows • Complete • Press “Open Door” • Using autoclave gloves and other PPE, remove and let cool to room temp. Autoclaves in SEH Building Autoclave Testing and Verification Bioindicators Most reliable is a live spore test using B. stearothermophilus Test packs using B. stearothermophilus are available from STERIS representatives. Place test pack on bottom shelf; run on a typical cycle. Should be tested monthly Autoclaves in SEH Building Autoclave Testing and Verification Prevacuum Efficiency DART – Daily Air Removal Test • Proprietary test pack – contact STERIS representative. • Cycle Select –test cycles- DART warmup first • Then, Cycle Select –test cycles- DART Leak Test • Should be run weekly • Cycle Select –test cycles-leak test • The leak rate will print out • Should be no more than 1mmHg per minute. Questions? Contact the Office of Laboratory Safety [email protected] 202-994-8258 Office of Laboratory Safety-OVPR The George Washington University 2300 Eye St. NW, Ross Hall Room B05 Washington, DC 20037 .