AUTOCLAVE: STANDARD OPERATING PROCEDURE (SOP)

Facility: National Laboratory

SOP Title: Autoclave: Operation and Maintenance SOP

Document Number: 3-02-006 Version Number: 01

Process Leader: Biorisk Management Advisor Effective Date: 01-01-2016

Other documents cross-referenced in this SOP (i.e., manuals, SOPs, forms, records):
• Biorisk Management Manual (Chapter V, Risk Assessment; Chapter XII, Laboratory Equipment;
and Attachment B, Protocol Risk Assessment Form) (4-00-001)
• Waste Disposal SOP (4-00-001)
• Personal Protective Equipment SOP (4-00-001)

Revision Sections Description of Change Date Approved

Number Changed By
SOP Title: Autoclave: Operation and Maintenance SOP

Document Number: 3-02-006 Version Number: 00

Effective Date: MM-DD-YYY

I Purpose
The purpose of this document is to establish procedures for proper use and maintenance of
autoclaves used by the National Laboratory. Adherence to this procedure allows the autoclave
to function as designed in order to provide the appropriate level sterilization and/or
decontamination of laboratory supplies and waste.

II Scope
This document applies to all laboratory personnel who use autoclaves within the National
Laboratory and is used when determined necessary by risk assessment to provide the
appropriate level sterilization and/or decontamination of laboratory supplies and waste.

III Responsibilities
• Process Leader ensures autoclaves are properly selected, located, operated and maintained and
that users are trained on this procedure.
• Laboratory personnel who use autoclaves follow the procedures outlined in this SOP and to
report any problems to the Process Leader.
• Facilities Management Office ensures that all autoclaves are installed, serviced, calibrated and
validated properly.

IV Preparation
A Materials
• Biohazard waste bags and containers
• Temperature indicator strips
• Steam sterilization indicator tape
• Biological indicator for steam sterilization
• Appropriate growth media or self-contained biological indicator and media
ampule
• Heat-resistant Gloves
• PPE as determined by risk assessment
• Mild detergent/cleaning agent
• “Out of Service” sign
B Equipment
• Autoclave: Tuttnauer 3850 EL Benchtop
C Records and Forms
• Autoclave Calibration Test Report
• Autoclave Validation Test Report
• Equipment Use Log

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SOP Title: Autoclave: Operation and Maintenance SOP

Document Number: 3-02-006 Version Number: 00

Effective Date: MM-DD-YYY

V Procedure (refer to Attachment A, Autoclave: Operation and Maintenance SOP Template Flow
Chart)

A Routine Operations
1.1 Set-up procedures and cycle selection
(1) Pre-heat
(a) At the beginning of each work shift, sterilize the first batch in a pre-heated sterilizer.
To do this, purge the sterilization chamber without loading material in the chamber.
(i) Close and secure autoclave door
(ii) Run program 18 (60˚C for 15 minutes)
(iii) Record on Equipment Use Log

(2) Glass sterilization

(a) Load glassware into autoclave within capacity limits. DO NOT OVERLOAD.
(i) Erlenmeyer Flask Capacity
1. 18 – 250 mL flasks
2. 11 – 500 mL flasks
3. 7 – 1000 mL flasks
4. 4 – 2000 mL flasks
5. 2 – 3000 mL flasks
(ii) Schott Duran Capacity
1. 24 – 250 mL flasks
2. 17 – 500 mL flasks
3. 12 – 1000 mL flasks
4. 7 – 2000 mL flasks
(b) Close and secure autoclave door
(c) Run program 1 (134˚C for 3 minutes)
(d) Record on Equipment Use Log
(e) Put on heat resistant gloves with long cuffs. Wear eye protection.
(f) Open door cautiously to allow remaining steam to vent without causing burns.
(g) Remove glassware

(3) Solid Waste Disposal

(a) Don appropriate PPE for safe handling of material to be autoclaved
(b) Place small amount of water into autoclavable waste bag with waste
(c) Include indicator test strip at least once a week. Place indicator within the bag in a
central location.
(d) Loosely seal waste bag with autoclave tape
(e) Load bag into autoclave, being careful to keep bag from touching the walls of the
autoclave.

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SOP Title: Autoclave: Operation and Maintenance SOP

Document Number: 3-02-006 Version Number: 00

Effective Date: MM-DD-YYY

(f) Ensure bag does not covering the chamber drain (pin trap).
(g) Close and secure autoclave door
(h) Run Program 4 (121˚C for 30 minutes)
(i) Record on Equipment Use Log
(j) Put on heat resistant gloves with long cuffs and other PPE as required by risk
assessment. Wear eye protection.
(k) Open door cautiously to allow remaining steam to vent without causing burns.
(l) Remove bag and check indicator strip.
(m) Follow Waste Disposal SOP for removal from lab and final disposition.

(4) Liquid Waste Disposal

(a) Don appropriate PPE for safe handling of material to be autoclaved.
(b) Ensure liquid is in an unsealed autoclavable container
(c) Place container in center of autoclave
(d) Close and secure autoclave door
(e) Run Program 4 (121˚C for 30 minutes)
(f) Record on Equipment Use Log
(g) Put on heat resistant gloves with long cuffs and other PPE as required by risk
assessment. Wear eye protection.
(h) Open door cautiously to allow remaining steam to vent without causing burns.
(i) Remove container.
(j) Follow Waste Disposal SOP for removal from lab and final disposition.

2.1 Safety considerations and work practices

(1) Always use heat resistant gloves with long cuffs to avoid burns.
(2) Do not tamper with autoclave during cycles.
(3) Beware of steam and pressure hazards.
(4) Always address alarms and follow maintenance protocols.
(5) Document use with Equipment Use Log.
(6) Ensure safety valves are not blocked.

3.1 Removal of materials and performance verification

(1) Routine Validation
(a)
(2) Performance Verification
(a)

B Scheduled Maintenance

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SOP Title: Autoclave: Operation and Maintenance SOP

Document Number: 3-02-006 Version Number: 00

Effective Date: MM-DD-YYY

(1) Autoclaves are calibrated/validated/serviced to what standard, at what frequency (daily,

weekly, monthly, quarterly, annually) and by whom (individual qualifications)?
(2) Describe manufacturer’s instructions for maintenance
(3) Are above service requirements different for autoclaves used for waste treatment (refer
to Waste Disposal SOP)?
(4) Are there service stickers and/or test reports?
(5) What records are maintained? By whom?

C Unscheduled Maintenance

(1) Is “out of service” signage used?

(2) Who is notified of autoclave deficiencies and unscheduled maintenance (Process Lead,
Facilities Maintenance Office)?
(3) What records are maintained? By whom?

VI References
A Tuttnauer Technical Specifications for Benchtop and Vertical Laboratory Autoclaves
B Autoclave Video Clip (QR Code on cover page)
C Manufacturer’s Instructions

VII Attachments
A Autoclave: Operation and Maintenance SOP Template Flow Chart

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SOP Title: Autoclave: Operation and Maintenance SOP

Document Number: 3-02-006 Version Number: 00

Effective Date: MM-DD-YYY

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