Environmental Services

Autoclave Efficacy Testing Procedure

Introduction to the Tools:
A number of tools are available to assess the performance of the autoclave; these include
physical, chemical and biological indicators. It is important to note that these indicators will only
respond to time, temperature and moisture conditions, and not to organic load.

Physical Indicators:

Pressure and temperature recording devices. Thermocouples can be

placed inside the load to determine the temperature achieved in the bag
itself.

Chemical Indicators: These indicators change colour after being exposed to specific
temperatures. There are two types of chemical indicators, temperature
sensitive tape (autoclave tape) and integrated chemical indicator strips.
For autoclave tape, the colour change will occur upon exposure to the
given temperature; it is not time related. Therefore these indicators can
only attest to the temperature attained and not to exposure time and
hence success of sterilization. Integrated chemical indicator strips
however, provide a limited validation of temperature and time, though
this is not as accurate as a Biological Indicator, and so it cannot be used
as a replacement.

Biological Indicators: Biological indicators are used in the efficacy testing of the autoclave
process to effectively sterilize the contents being treated.
G.
stearothermophilus spores are used, as they are the most resistant
organism to steam autoclaving. To determine the effectiveness of the
autoclave process the biological indicator must be placed in a typical test
load (solid or liquid) and exposed to the typical cycle conditions. This is
the standard method of validating the effectiveness of your autoclave
procedures. Testing using a biological indicator must be undertaken
monthly, unless otherwise authorized by RMS.

Revised by: Stephanie Thomson

Date: July 29, 2013

Risk Management Services

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Environmental Services

Procedure for Efficacy Testing with Biological Indicators:

1. Read and follow the suppliers instructions. Link to Instructions.
2. Tie a string around the BI tube and place the G. stearotheromophilus in centre
of representative test load (inside a waste bag).
3. Process load in normal fashion.
4. Extract and process the BIs
a. Pull the BI from the waste bag and remove the string
b. Using the ampule crusher, break the glass ampule inside the plastic
tube. Be careful not to break the plastic. Do this for the positive
control (a tube that did not go into the autoclave) and the autoclaved
BI.
c. Incubate G.stearothermophilus samples as instructed by manufacturer
(55-60oC for 24-48hrs).
5. Check for colour change at regular intervals during the
incubation period (24, and 48 hours). If media is yellow and
turbid the autoclave process has FAILED. Immediately upon
noting yellow colouration, re-run all samples with new biological
indicators.
6. If failure continues to be noted, either increases time of
exposure or initiate repairs to the autoclave. Note the autoclave
cannot be used again until validations procedure indicates that
autoclave is now adequately sterilizing the material.
7. Record all results. (Positive and Negative)
Records:
1. Record biological indicator information: brand, lot #, expiry date.
2. Record the date, operator, cycle time, cycle temperature of the test run.
3. Record results: colour change noted - yellow (Failure), no colour change purple (Pass)
4. Email the results to autoclave.report@ubc.ca

Revised by: Stephanie Thomson

Date: July 29, 2013

Risk Management Services

Page 2