STANDARD OPERATING PROCEDURES 

Safe Autoclave Operations



     The purpose of this document is to provide standard operating procedures for the
safe use of autoclaves. Autoclaving is a process used to destroy microorganisms and
decontaminate biohazardous waste and microbiological equipment used at
Biosafety Level 1 / 2.
Autoclave Make/Model:
EX:  •Consolidated Stills & Sterilizers, Model # SSR3APB
Serial #42090
Person Responsible and Phone #: ______________
Located in Building/Room: Building ____, room ____


HAZARDS

      Autoclaves use high pressure (15 psi) and high temperature(121 C) or 250 F)  steam for sterilization.
Potential safety risks for the operators include:
          · Heat burns from hot materials and autoclave chamber walls and door.
          · Steam burns from residual steam coming out from autoclave and materials
          on completion of cycle.
          · Hot fluid scalds from boiling liquids and spillage in autoclave.
          · Hand and arm injuries when closing the door.
          · Body injury if there is an explosion.

SAFETY

     To insure the health and safety of personnel using the autoclave, it is important for each department to maintain autoclaves and train personnel annually in there  proper use
          · The name of the person responsible for the autoclave shall be posted near the
          autoclave. This SOP should be posted on the outside to the autoclave.
          · It is the supervisor's responsibility to ensure employees are trained before
operating any autoclave unit.  Training should occur annually after that.
          · Procedural and instructional documents provided by the manufacturer must
          be followed.
          · Personal protective clothing and equipment must be worn when loading and
          unloading the autoclave.
         · Autoclaves preventative maintence (PM) service performed at least 3 times a year.
         A basic visual inspection should be performed monthly by the person responsible
         for the autoclave. The inspection, service and repair records should available upon
         request and all reports are to be kept in the autoclave log book.

          -Spore strips, from SPI, may be used to validate autoclave effectiveness. They
         should be done once a month and sent to the company. Results should be put in
         autoclave log book.

PERSONAL PROTECTIVE EQUIPMENT Equipment to protect against scalds and
burns include:
          · Heat-insulating gloves that provide complete coverage of hands and forearm
         · Lab coat
         · Eye protection: indirect vented tight sealing goggles
          - Closed-toe footwear
          - long pants

OPERATOR INSTRUCTIONS

Training
     All personnel who use autoclaves must have successfully completed an annual
training session from their supervisor or safety officer on the safe operating
procedures. This requirement applies to both new and experienced personnel. A
record should be kept for inspection purposes.

Material Preparation
Ensure that the material is safe for autoclaving:
          · Samples containing solvents or substances that may emit toxic fumes should
          not be autoclaved. Example: formaldehyde.
          · Do not autoclave bleach!
         - Glassware must be inspected for cracks prior to autoclaving.

Prepare and package material suitably:
         · Loose dry materials must be wrapped or bagged in steam-penetrating paper or
          loosely covered with aluminum foil. Wrapping too tightly will impede steam
          penetration, decreasing effectiveness of the process.
         · Loosen all lids to prevent pressure buildup. All containers must be covered
         by a loosened lid or steam- penetrating bung and placed in a tray as double
         containment in case of a spill. Liquid must be in an autoclavable container with a
         secure lid backed off ¼ turn, if doing flasks then seal with foil to maintain sterility.
         · Containers of liquid must not exceed two-thirds (2/3) full, with lids loosened.
         · Glassware must be heat-resistant borosilicate, chip and or crack free
         · Plastics must be heat-resistant, i.e., polycarbonate (PC), PTFE (“Teflon”) and
         most polypropylene (PP)items.
         · All items must be tagged with autoclave tape.

Place items in secondary containers to secure and contain spills:
         · Items should be placed in a stainless steel pan or other autoclavable container
          for their stability and ease of handling.
         · Place containers of liquid, bags of agar plates, or other materials that may boil
          over or leak, into a secondary pan in the autoclave.
         · The pan must be large enough to contain a total spill of the contents.
         · Bags must not be tightly sealed as steam cannot penetrate.

Biohazard Waste: 

    State law requires all biohazardous waste be autoclaved or picked up within 7 days, once it exceeds 20lbs

         - An autocalve load must be run prior to running a waste load to ensure the
          autoclave is working properly.
          - All waste must be weighed, recorded in the log book and sterilizer strips run
         with the waste log. Seterilizer strips need to be stapled to the log book as well as the
         temp log.
         - Once a month: Run SPS strips and send to the company for sterilization
         verification.
        - Waste permit issued by the city must be posted next to the autoclave.
         We are a small waste generator and can not exceed 200lbs. If the permit is outdated
         contact safety officer:

- **Once a year the autoclave thermometer must be calibrated and
documentation posted by the autoclave as well as put in the
autoclave book by a licensed technician**

Bio Haz Waste:
    Place items in secondary containers to secure and contain spills:
          · Items should be placed in a stainless steel pan or other autoclavable container
           for their stability and ease of handling.
          · Place containers of liquid, bags of agar plates, or other materials that may boil
           over or leak, into a secondary pan in the autoclave.
          · The pan must be large enough to contain a total spill of the contents.
          · Bags must not be tightly sealed as steam cannot penetrate.

Loading Autoclave

     · Wear lab coat, eye protection, heat-insulating gloves, and closed-toe shoes. ·
Place material in autoclave. Do not mix incompatible materials.
     · Do not overload; leave sufficient room for steam circulation. If necessary, place
     the container on its side
     to maximize steam penetration and avoid entrapment of air.
     · Close and latch door firmly.

Operating Autoclave

     · Close and lock door.
     · Choose appropriate cycle (e.g., gravity, liquid, or dry cycle) for the material.
     Consult the autoclave manual for assistance in choosing a cycle. The manuals for
     operation of the autoclave should be located near the autoclave.
     · Set appropriate time and temperature if you are using a customized cycle.
     · Start cycle and fill out the autoclave user log with your contact information. A
     completed cycle  usually takes between 1-1.5 hours, depending on type of cycle.
     · Do not attempt to open the door while autoclave is operating.
     · If problems with your autoclave are perceived, abort cycle and report it to
     your PI/Lab manager immediately.

Unloading Autoclave

     · Wear heat-insulating gloves, eye protection, lab coat, long pants and closed-toe
     shoes.
     · Ensure that the cycle has completed and both temperature and pressure have
     returned to a safe range.
     · Wearing Personal Protective Equipment (PPE), stand back from the door as a
     precaution and carefully open door no more than 1 inch. This will release residual
     steam and allow pressure within liquids and containers to normalize.
     · Allow the autoclaved load to stand for 10 minutes in the chamber. This will
     allow steam to clear and trapped air to escape from hot liquids, reducing risk to
     operator.
     · Do not agitate containers of super-heated liquids or remove caps before
     unloading.  They can boil over explosively!
     · Wearing heat-insulated gloves remove items from the autoclave and place
     them in an area which clearly indicates the items are ‘hot’ until the items cool to
     room temperature.
     · Shut autoclave door.

Autoclave Use Log

     · Entries must be placed in the log book each time the autoclave is used. These
     records are used for maintenance/service schedules and reporting of incidents,
     accidents and/or faults.
     · Entries should include: operator's initials, date, time and duration.
     · The log book must be kept adjacent to the autoclave.
     · An Autoclave Use Log example is provided in this document.


Maintenance and Repair

     · No person shall operate the autoclave unless the autoclave is in good repair.
     · Only qualified professionals are permitted to make repairs.
     · Report possible malfunctions to Tasha Sturm
     · Repairs are performed by your service contract or any other contractor you
     choose to hire.

CONTINGENCY PLANS

Equipment Malfunction
     · If the autoclave does not operate exactly as expected, do not attempt to fix the
     problem. A notice shall be placed on the autoclave indicating that it is not to be used
     until the problem is diagnosed and corrected.
     · Record the problem in the autoclave log book.
     · Contact or your supervisor to report the problem. ·
**Only qualified professionals are permitted to make repairs.**

Incident Response

     · All incidents, including a spill or release of biohazardous materials and
     recombinant or synthetic nucleic acid molecules, must be reported to your
     supervisor.
     · If any injury occurs seek first aid or, if necessary, seek medical assistance at
     the  Heath Services or by dialing 911.
     · If clothing is soaked in hot water/steam, remove clothing and place the injury
     in cool water.
     · Place a notice on the autoclave indicating that it is not to be used
     until the cause of the incident is determined, procedures enacted to prevent future
     incidents, and the autoclave is deemed safe for operation.


Spill Clean-up

     · Spills may occur from a boil-over or breakage of containers.
     · No operation of the autoclave is allowed until the spill is cleaned up.
     · The operator is responsible for clean-up of spills. Contain the spilled material
     using paper towels. Use your laboratory’s spill kit if necessary. Wait until the
     autoclave and materials have cooled to room temperature before attempting clean-
     up.
     · Review the Safety Data Sheet if appropriate, to determine appropriate PPE,
     spill cleanup and disposal
     protocols that are necessary.
    · Dispose of the waste following the protocol appropriate for the material (e.g.,
     red biohazard bag). If materials have been intermingled, follow the clean-up and
     disposal protocol for the most hazardous component of the mixture.
    · Cracked glassware must be disposed of properly.
    · Record the spill and clean-up procedure in the autoclave log book.

Document modified from Arizona State University,

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