Procedures for Using Steam Autoclaves QUESTIONS: Contact DRS, 217-333-2755, for additional information. CHOOSE PROPER OPERATING CONDITIONS: MAINTENANCE & CARE: DECONTAMINATING BIOHAZARDOUS Check the Pressure: • Sterilization will occur only when the conditions of time, temperature, pressure, and humidity have been met. WASTE BAGS: Check all pressure gauges. • Incorrect selection of time or exhaust cycle may damage the autoclave, cause liquid to boil over, or bottles to break. • Use autoclavable, high strength Jacket pressure gauge should be a • Proper use of autoclave will minimize the chance of serious injury. polymer bags imprinted with minimum of 15 psi (refer to Biological Waste Liquids Dry Items Glassware chemical indicator (if possible). Items manufacturers’ instructions for (Gravity Cycle) (Liquid Cycle) (Gravity Cycle) (Gravity Cycle) • Add approximately 250 mL water to maximum psi). Loosen caps or use a Dirty: Place in middle of bag before closing and transporting Open the bag >2", Fabrics Wrap; vented closure, the pan; to autoclave area. Use Sterilization Indicators Preparation Place in tray, Instruments: Clean, Fill containers no more Clean: wash, rinse, Place indicator if needed dry, lay in pan • Place bag in a polypropylene or Visual indicators, e.g., than 75% capacity wrap stainless tray before autoclaving. chemical/tape indicators, measure Dirty: In detergent and Fabrics: Separated, on pan; • Open the bag at least 2“ to allow one or more physical conditions of Placement in Autoclave In the center Upright in pan edge; Clean: On side or the autoclave cycle. Instruments: Flat steam to enter. inverted Mechanical indicators record the • When complete, the sterilization time-temperature profile attained indicators must show adequate Temperature 121oC 121oC 121oC 121oC during a cycle. decontamination before disposal as Biological indicators, i.e., spore municipal solid waste. 22 min. for volumes 60-120 min. • Repeat cycle if sterilization vials, are the only approved way to Treatment Time in <100mL; depending on load size 30-60 min. 30-60 min. Minutes 40 min. for volumes indicators do not show evidence of prove sterility. and packing density >100mL sterilization. NOTIFY FACILITIES SUPPORT • Overbag biohazardous waste bag Dirty: Slow exhaust; Exhaust Cycle Slow exhaust Slow exhaust Fast exhaust and dry with an opaque trash bag before WHEN A PROBLEM OCCURS: Clean: Fast/dry No steam. placing in the regular trash. Valves leaking. Avoid puncturing bags. Check reference for Glassware with cracks or Hot bottles may explode. Door gasket deterioration. Notes: Overbag and dispose of proper packaging deep scratches may Let cool before moving. Reminders properly. methods crack Jacket pressure below 15 psi. • Store biohazardous waste in a Temperature not reaching 121°C CAUTION: closed leak-proof container. (250°F). • NEVER AUTOCLAVE FLAMMABLE, REACTIVE, CORROSIVE, TOXIC or RADIOACTIVE MATERIALS, e.g., bleach. • Never allow waste to accumulate in Erratic temperature or pressure • Materials that melt (plastic lab wear) at ≥ 121°C will block chamber exhaust drain if not placed in a shallow the lab. during cycle. autoclave pan able to withstand that temperature. • Never leave waste unattended. Steam escapes from around door • Use caution when increasing autoclave temperature to 135oC because plastics (including some plastic pans) • Clean up leaks and spills with a during cycle. melt at this temperature, causing difficult clean-up and damage to temperature sensors. suitable disinfectant. Water on floor or in chamber after • Always wear safety glasses, goggles, or face shield, lab coat or apron, and heat-protective non-asbestos a cycle. gloves when opening door or removing item(s) from autoclave. Articles very wet after a fast • Do not mix loads that require different exposure times and exhaust. Reference: “Using the Gravity exhaust or dry cycle. • Open door only after chamber pressure returns to zero. Leave door open for several minutes to allow Displacement Steam Autoclave in the Excessive steam in area during pressure to equalize and for materials to cool. Biomedical Laboratory” cycle or after opening door. • Open door slowly. Beware of rush of steam or water. DHHS/PHS/HIH/DS .