Autoclave Operation

Specific Procedures

The Biosafety Office has detailed instruction sheets for UF’s more heavily used autoclaves. Click on the applicable link below. The instruction sheets are NOT  SUBSTITUTES for PROPER TRAINING. Schedule a training session by contacting our office.


Obtaining Autoclave Access Steris Century Models in Academic Research Building (ARB)

This video shows how to obtain access in order to use the Steris Century Autoclave units


Obtaining Autoclave Access Steris Lab 250 Models in Academic Research Building (ARB)

This video shows how to obtain access in order to use the Steris 250 Autoclave units


Recommended Sterilization Times [Drying Time not Included]

121ºC / 250ºF & minimum 15PSI (Gravity Cycle)

  • BIOWASTE IN AUTOCLAVE BAGS, LOOSELY CLOSED/VENTED
    • MULTIPLE BAGS ― 100 MINUTES OR LONGER
    • SINGLE BAG ― 90 MINUTES
    • PARTIAL BAG ― 60 MINUTES
  • DRY GOODS
    • GLASSWARE, EMPTY, INVERTED ― 15 MINUTES
    • INSTRUMENTS, WRAPPED ― 30 MINUTES
    • UTENSILS, WRAPPED ― 30 MINUTES
  • LIQUIDS (BOTTLES WITH VENTED CAPS, ½ FULL
    • 75mL ― 25 MINUTES
    • 250mL ― 30 MINUTES
    • 500mL ― 40 MINUTES
    • 1000mL ― 45 MINUTES
    • 1500mL ― 50 MINUTES
    • 2000mL ― 55 MINUTES
      Please note that Liquid cycles will take up to an additional 30 minutes to run than the set sterilization time due to slow charge and exhaust.

Record Keeping

The following records regarding autoclave use must be kept:

  • Maintenance records
  • Autoclave use log – must be available near the autoclave 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?

Testing

Autoclaves shall be tested before being placed into service and then periodically to ensure they are functioning as required:

  • 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 or sterilize other material.)

A commercially available test indicator kit that uses bacterial spores (Geobacillus stearothermophilus) is the approved method of testing autoclave efficacy. Most spore vial test kits require 560 to 600 C incubation of the autoclaved test vial along with a non-autoclaved control vial. Incubation causes surviving spores to grow, indicated by a color change. 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. 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. For periodic testing, place a spore vial in the very center of a test load prior to autoclaving.

Please contact EH&S with questions on autoclave testing or to check if your autoclave is on the schedule for testing by EH&S.