Biological Safety

Autoclave Use and Testing

Rules governing the use and testing of autoclaves are based on Rule 64E-16 of the Florida Administrative Code.

Autoclave Testing
Autoclaves shall be tested before being placed into service and then periodically for effectiveness.

1. Periodicity
• Every 40 hours of use (Required for autoclaves that are used to inactivate human or non-human primate blood, tissues, clinical samples, or human pathogens.)
• Every 6 months (Required for autoclaves that are used to inactivate other material.
2. Method
• A commercially available test indicator kit that uses bacterial spores (Bacillus stearothermophilus) is the approved method of testing autoclave efficiency. Most spore vial test kits require 56 to 60 ° C incubation of the autoclaved test vial along with a non-autoclaved control vial. Incubation causes surviving spores to grow.
• New autoclaves
  Before placing an autoclave into service, a test load approximating the weight and density of the type of waste generated shall be autoclaved with test spore vials. The spore vials should be placed at the bottom, top, front, rear, and center of the autoclave chamber. This can be achieved by either:
• placing vials at those positions within one large test load, OR
• making several smaller test packs with 1 vial at the center of each and placing the packs at those locations within the chamber.

The appropriate parameters for sterilization including temperature, pressure, and treatment time shall be determined in this way.

• Autoclaves already in use
  For periodic testing, place a spore vial in the very center of a test load prior to autoclaving.
2. Storage Information
• Please read the product information sheet for appropriate storage information, but, in general, spore vials should not be frozen. Each batch of vials has an expiration date. Vials should not be used after their expiration date.

 
Record Keeping
The following records regarding autoclave use must be kept:

1. On-site maintenance records
2. Autoclave use log (Each load of material inactivated shall be logged as follows:)
• Date, time, and operator's name
• Contact information:  Lab, room number, phone number
• Is this biohazardous material?
• Record the temperature, pressure, and length of time the load is sterilized. Please note that temperature sensitive autoclave tape is not sufficient to indicate that the load reached sterilization conditions because the tape will change color at lower temperatures
• Save the autoclave print-out, if the autoclave has a working printer.
   

Autoclave Operating Procedures
A written sterilization procedure shall be in place for each workplace. This shall include the following:

1. Parameters
• Appropriate parameters for sterilization shall be determined from the testing with spore vials.
• The time it takes to sterilize a load will change depending upon the load density and the sterilization cycle one chooses. Tests have been performed which imitate these various situations. Please follow the established guidelines.
2. Protocol
• Identification of standard treatment containers and proper load placement shall be made.
3. Cleaning
• The autoclave and work areas shall be cleaned after every use and the work area shall be disinfected, as needed.
   

Autoclave Operation and Safety Training
Autoclave training is given by the EH&S Biological Safety Office both quarterly and upon request. The training is geared toward the research staff. It goes over proper use of autoclaves and how they may be maintained and used properly. Safety training is also given.

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