Autoclaves Proper Operation and Validation

By Tom Holm, esq., BME Labstore

August, 2011

Is your autoclave validation current? Probably. Do your operating procedures and

validation reports satisfy the new requirements adopted in 2009 by the MN Department of
Healths environmental lab accreditation program? Ahhh, not sure? After discussing
autoclave operation and validation requirements with a number of laboratory personnel, it is
evident that many are not familiar with these new requirements and, furthermore, do not
know what tests are available to satisfy the requirements.
What new requirements? Effective July 1, 2009, in anticipation of implementing a
nationally accredited program, the MN Department of Health adopted the National
Environmental Laboratory Accreditation Programs (NELAPs) lab standards, NELAC 2003,
as the governing standard for MN environmental lab accreditation. As a result, MN
Administrative Rule 4740.2110 subparagraph 11 (a copy of NELAC 2003, Appendix D,
D.3.8.2) now governs the operation and validation of autoclaves used by accredited labs for
bacteriological testing. In September 2010, Minnesota completed this process and became
the 15th state to be approved by NELAP.
Specifically, what does the Rule require? To summarize, Rule 4740.2110 subp. 11
requires the following:
1. The performance of each autoclave must be evaluated initially by establishing its
functional properties and performance, for example heat distribution characteristics
with respect to typical uses.
Functional properties and performance characteristics would require tests to
determine:
a.
b.
c.
d.
e.
f.
g.
h.

How long does it take to reach sterilization temperature (250 0F)? Does this
differ under a load?
Are there any cold or hot spots within the autoclave?
Is it able to hold the target temperature with minimal temperature variations?
Does the internal temperature coincide with the temperature on the display,
gauge or dial?
Is there adequate pressure? Is there too much pressure?
Is the timer accurate? Does the timer start at the beginning of the cycle or
upon reaching the target temperature?
Is the autoclave able to hold a sufficient amount of water to complete the
cycle?
What cycle time is required to achieve adequate sterilization of a typical
load?
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Why require this information? These performance characteristics enable you

and the auditor to determine if the autoclave is performing adequately. In
addition, in the case of a poorly performing autoclave, these performance
characteristics can also help diagnose specific problems. For example, a cold
spot might indicate a poorly performing heating element or a leaky door gasket.
A long come-up time may indicate a clogged venting tube or a bad heating
element. A sharply rising temperature may indicate an insufficient water supply.
A discrepancy in temperature may indicate a bad temperature controller. Most
importantly, a comprehensive picture of the autoclaves performance
characteristics provides a baseline for comparison against future tests to
determine if the autoclave is still performing as it should.
2. Demonstration of sterilization temperature must be provided by use of a continuous
temperature recording device or by use of a maximum registering thermometer with
every cycle. Appropriate biological indicators must be used once per month to
determine effective sterilization. Temperature-sensitive tape must be used with the
contents of each autoclave run to indicate that the autoclave contents have been
processed.
New rules require autoclaves to be equipped with either a printer or maximum
registering thermometer and used in conjunction with temperature-sensitive tape for
each sterilization cycle. A log of the results must be kept. While a mercury-filled
thermometer is not prohibited by state statute if it is used to calibrate other
thermometers or equipment, it certainly poses additional, potential risks for the
laboratory.
3. Autoclave maintenance, either internally or by service contract, must be performed
annually and must include a pressure check and calibration of the temperature
device.
A pressure test and temperature calibration must be performed annually utilizing
instruments traceable to NIST. While a comprehensive temperature distribution test
is not required annually, it is still highly recommended because it enables you to
compare performance characteristics against earlier tests and identify current and
future problems.
4. Additionally, The autoclaves mechanical timing device must be checked quarterly
against a stopwatch and the actual time elapsed must be documented.
Proper Validation
The best way to meet the new requirements and perform a proper validation is to
utilize temperature and pressure data loggers to accurately monitor and record the
temperature and pressure throughout a complete cycle. With a proper mapping, all of
the functional properties and performance characteristics can be recorded.

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Following is an example of a validation performed with data loggers. Figure 1 shows

the mapping of the data loggers within the autoclaves chamber. A proper mapping is
determined by the location of the door, the heating elements, vents, temperature
sensors, and shape of the autoclave.

Figure 2 is a graph showing the temperature throughout a sterilization cycle. From this
graph, it is possible to determine the following performance characteristics:
a. Come-up time for the cycle was approximately 20 minutes.
b. There does not appear to be any hot or cold spots as indicated by the consistency of
all of the probes.
c. The autoclave was able to hold the target temperature with minimum fluctuations in
temperature throughout the cycle.
d. The autoclaves timer, temperature, and pressure gauges can be validated against the
certified data logger data.
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Figure 3 is a graph showing the pressure throughout a sterilization cycle. From this graph, it
is possible to determine the following performance characteristics:
a. The autoclave was able to reach and hold an acceptable pressure level.
b. There was more initial pressure variation in the back of the autoclave than in the
front. This variation was caused by the probe being closer to the vent.
c. The vent appears to be working properly. Air and steam are able to be exhausted at
the beginning, throughout and end of the cycle.

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Figure 4 is a graph showing the F values or heat equivalency throughout a sterilization cycle.
From this graph, it is possible to determine the following performance characteristics:
a. An F value of 8 (heat equivalent of 8 minutes at 250 0F) was reached after about 10
minutes. An F value of 8 is normally adequate to sterilize glassware and stainless
utensils. This information can be helpful in determining the required cycle time for
adequate sterilization.
b. The F values will vary greatly depending on the size and content of the load. A
validation test with a typical load provides more accurate data for determining
adequate sterilization time and temperatures.

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Summary. As of July 1, 2009, the Minnesota Dept. of Healths Environmental Lab

Accreditation Program (MN-ELAP) requires more stringent testing of autoclaves to ensure
adequate sterilization procedures. These requirements are best satisfied using certified data
loggers that are able to record temperature and pressure throughout the sterilization cycle.
Comprehensive tests enable the operators and auditors to determine the autoclaves
functional properties and performance characteristics. Monitoring these performance
characteristics also enables the autoclave operator to recognize changes in the autoclaves
performance and provide preventive maintenance services prior to sterilization failures.

BME Labstore provides lab equipment sales and service throughout Minnesota and western Wisconsin. For
questions or comments about this article, you may contact Tom Holm @ 651-646-5339 or
tom.holm@bmelabstore.com.

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NELAC 2003
QUALITY SYSTEMS APPENDIX D
ESSENTIAL QUALITY CONTROL REQUIREMENTS
D.3

MICROBIOLOGYTESTING
D.3.8 Constant and Consistent Test Conditions
a)

LaboratoryFacilities

Floors and work surfaces shall be non-absorbent and easy to clean and disinfect.
Work surfaces shall be adequately sealed. Laboratories shall provide sufficient storage
space, and shall be clean and free from dust accumulation. Plants, food, and drink shall be
prohibited from the laboratory work area.
b)

LaboratoryEquipment
1)

TemperatureMeasuringDevices

Temperature measuring devices such as liquid-in-glass thermometers,

thermocouples, and platinum resistance thermometers used in incubators, autoclaves and
other equipment shall be the appropriate quality to meet specification(s) in the test method.
The graduation of the temperature measuring devices must be appropriate for the required
accuracy of measurement and they shall be calibrated to national or international standards
for temperature (see 5.5.6.2.2.2). Calibration shall be done at least annually.
2)

Autoclaves

i) The performance of each autoclave shall be initially evaluated by establishing its

functional properties and performance, for example heat distribution characteristics with
respect to typical uses. Autoclaves shall meet specified temperature tolerances. Pressure
cookers shall not be used for sterilization of growth media.
ii) Demonstration of sterilization temperature shall be provided by use of continuous
temperature recording device or by use of a maximum registering thermometer with every
cycle. Appropriate biological indicators shall be used once per month to determine
effective sterilization. Temperature sensitive tape shall be used with the contents of each
autoclave run to indicate that the autoclave contents have been processed.
iii) Records of autoclave operations shall be maintained for every cycle. Records shall
include: date, contents, maximum temperature reached, pressure, time in sterilization
mode, total run time (may be recorded as time in and time out) and analysts initials.
iv) Autoclave maintenance, either internally or by service contract, shall be performed
annually and shall include a pressure check and calibration of temperature device.
Records of the maintenance shall be maintained in equipment logs.
v) The autoclave mechanical timing device shall be checked quarterly against a stopwatch
and the actual time elapsed documented.

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