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The University of Tennessee

Office of Research -- Biosafety Compliance

Frequently Used Tools:




Autoclave Validation Instructions

  1. Place a full, medium-sized biohazard bag (e.g. 25" x 35") into an autoclavable secondary container (i.e. autoclave pan).
     
  2. Add 1 cup (~250ml) WARM water to the bag. This is recommended for all bagged waste, but it is REQUIRED if the contents are not likely to generate steam within the bag (e.g. bag contains mostly dry plastic disposables).
     
  3. Place a 3M Comply Thermalog Steam Chemical Integrator strip inside the bag near the center of the bagged contents. Make sure that there is a mechanism for retrieving the strip after the validation test. For example, affix a piece of twine to the strip and tape one end of the twine to the outside of the bag. The Biosafety Office will provide the test strips (twine included) as requested.
     
  4. Place bag into the autoclave; LEAVE THE TOP OF THE BAG OPEN as adequate sterilization conditions may not be achieved within a closed bag. Helpful hint: make sure that the bag is opened widely at the top (i.e. do not bind/tie or twist the top) and that the opening is not obstructed to accommodate the dimensions of the autoclave (i.e. folding the top over or "cramming" the bag against the walls/ceiling).
     
  5. Autoclave the biohazard bag for a minimum of 30 minutes at 250F/121C. Helpful hint: 30 minutes is the recommended minimum, but sterilizations of >1 hour are not abnormal depending on the autoclave and load volume/contents.
     
  6. After cycle completion, note the status of the chemical integrator and please fill out the information below (all fields are required):

Autoclave ID number:

An Autoclave ID number is required.

Building:

A building name is required.

Room Number:

A room number is required.

Sterilization parameters for successful test (temperature AND time):

Paramters are required.

Cycle identifier if available (e.g. Liquids 2 or Gravity 1):

A cycle identifier is required. Type N/A if not available.

Load contents (identify the types of waste in the bag; e.g. agar plates, disposable tissue flasks):

A description of load contents is required.

Validation method (3M integrator strip):

A validation method is required.

Date and time of validation:

A date and time of validation is required.

Person performing the test:

The name of the person performing the test is required.

Email of person performing the test:

An email address for the person performing the test is required.
Please enter the email address in the proper format.

Notes/comments including any failed attempts
(if none, please enter N/A):

If you do not have any comments, please type N/A.

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  1. If the validation was successful the biohazard bag may be tied up and discarded into a non-see-through trash bag for final disposal. If the validation was not successful, repeat the test and add 10 minutes to the sterilization cycle. Integrator strips are single-use only. Use a NEW strip for each retest.
     
  2. Enter the appropiate information on the Autoclave Validation Sign Form. Then print and post the Autoclave Validation Sign next to the autoclave. If the autoclave does not pass validation, print and post the Stop Sign next to the autoclave.

Disposal of Biological WastesUT Biohazardous Waste Basics: A Guide for Handling & Disposal of Biological Wastes Generated in the UT Research & Diagnostic Service Environment -- PDF

Autoclave

Contact Information

Biological Safety Officer
Brian Ranger
336 Ellington Plant Sciences
Knoxville, TN 37996
Phone: (865) 974-1938
Email: branger@utk.edu

Biological Safety Specialist
Sarah DiFurio
336 Ellington Plant Sciences
Knoxville, TN 37996
Phone: (865) 974-9836
Email: sbottoml@utk.edu