At MIT, autoclaves are used to sterilize waste and clean materials associated with biomedical research. The Autoclave Validation Program oversees compliance of biohazardous waste management at the state level (Massachusetts State Sanitary Code Chapter VIII 105 CMR 480.000) and assists in maintaining the institute’s international accreditation with AAALAC (Association for Assessment and Accreditation of Laboratory Animal Care). The program is managed by the EHS Biosafety Program, with local coordination and operational collaboration across the MIT campus.
The MIT Autoclave Validation Program performs challenge testing on all autoclaves used within MIT research facilities. Validation tests are performed with a small self-contained biological indicator (BI) containing a standard challenge population of Geobacillus stearothermophilus spores, a non-pathogenic thermophilic bacterium that is naturally resistant to high heat and pressure. Inactivation of G. stearothermophilus spores can be achieved with a 16-minute exposure to typical autoclave conditions of 121°C and 15psi, and as a result, these spores are an ideal biological indicator of microbial viability after autoclave sterilization.
The Autoclave Validation Program oversees compliance of biohazardous waste management and assists in maintaining the institute’s international accreditation with AAALAC (Association for Assessment and Accreditation of Laboratory Animal Care)
There are 4 cycles each year, and each autoclave on campus has to be validated. The validation process includes incubation of a Biological Indicator (BI) during a sterilization cycle, followed by overnight incubation of the BI at 65°C (either at the lab or by BSP staff)
Vials are available at the VWR room (56-070) and in the EHS office (N52-496). Vials can also be sent via interdepartmental mail. To request vials, please email BSP@mit.edu or contact your EHS coordinator.
The yearly validation cycle dates are:
Cycle number
Cycle dates
Cycle validation deadline
Q1
1/6 – 3/31
3/1
Q2
4/1 – 6/30
6/1
Q3
7/1 – 9/30
9/1
Q4
10/1 – 12/21
21/1
Autoclaved biological indicator (BI) may be dropped off in the VWR stockroom (56-070), returned to your BSP Officer, or returned directly to the EHS office (N52-495) in person or by mail. Returned vials will be processed by the BSP staff. Validators may complete the process in their own lab. See below for instructions.
Instructions for in-lab autoclave validation processing
Place the BI in the center of the load.
Select the cycle used for the sterilization of biohazardous waste. The cycle must include a minimum temperature of 121°C or 250°F for 30 minutes or longer, depending on size and compaction of the load. The full cycle time will take 60-90 minutes.
Remove the indicator when the cycle is complete and the load is cool enough to handle.
Process the autoclaved indicator and an indicator that has not been autoclaved (as a control) by first gently crushing the inner glass ampoule, this can be done by pressing on the outer plastic sheath near the brown cap; when broken you will see the purple media dampen the spore paper at the bottom of the vial. Then incubate at 55-60°C for 24 hours.
If growth occurs in the processed indicator, the autoclave has not inactivated the spores. Test the autoclave again with a new biological indicator and consider adding a secondary sterilization validation device like a chemical integrator. If, after a second attempt, the autoclave has not inactivated the spores, the autoclave must remain out of service until a successful biological indicator test is achieved. The person responsible must inform users not to use the autoclave and post a “Do Not Use” sign while the autoclave is out of service
Safety training on autoclave use
Please follow this autoclave use SOP to ensure the safe and effective operation of autoclaves in MIT research labs.
An online training on autoclave safety is available through the MIT Atlas Learning Center.
Go to the course catalog and search for “Autoclave Safety Training” (EHS00254w).
You also must complete an in-person training; this is typically done by your EHS Representative, experienced researcher, PI, or EHS coordinator.
Clean materials (glassware, Surgical tools, liquids, etc.) should be collected and placed in an autoclave-proof tray.
Run the appropriate cycle (121°C, 15 psi, 15 – 60 minutes).
After the run, allow the load to cool for at least 10 minutes.
Caution: Failure to allow sufficient cooling time may result in injury
Can I autoclave waste?
MIT disposes of biological/medical waste primarily using biowaste boxes,
Biological/medical waste is autoclaved under certain conditions (permit requirements, only effective method of waste inactivation, per specific approved risk assessment, etc.).
Please consult with your biosafety officer if you have questions or concerns.
How do I know if I can autoclave waste in a specific autoclave?
Autoclaves are designated and labeled “for waste.” Not all DLCs have autoclaves designated specifically for waste. If you are unsure whether you should autoclave your waste, please consult with your EHS coordinator or Biosafety contact.
Sterilization of waste
Follow the autoclave use SOP to ensure the safe and effective operation of the autoclave.
Waste should be collected and placed in a clear bag in a marked (biohazard labeled) transfer container.
For liquids, leave cap slightly OPEN for effective steam penetration during autoclaving and to prevent pressurization of the container and spills inside the autoclave.
Leave the bag OPEN for effective steam penetration during autoclaving.
Place waste bag in a secondary autoclave-proof container and run waste cycle (121°C, 15 psi, 60 minutes).
After the run, allow waste to cool for at least 10 minutes. Caution: The waste is hot. Wear appropriate PPE when handling hot waste
Failure to allow sufficient cooling time may result in injury.
Enter the waste run in the autoclave log binder, usually near the equipment.
Place treated and tagged waste bags into regular waste for custodial pickup.
Please consult your EHS coordinator or BSP@mit.edu for questions and concerns.
Autoclaves that are designated for waste must:
Autoclaves designated for waste must:
Be labeled “For Waste” on or near the autoclave. (Note: Not all DLCs have autoclaves specifically designated for waste. If you are unsure whether to autoclave your waste, please consult your EHS coordinator or Biosafety officer.)
Be updated in the QuickBase database as a waste autoclave. Please confirm with your EHS coordinator or Biosafety contact whether your autoclave is designated for waste.
Have a completed Autoclave Waste Log template placed nearby. This log must be retained for three years after the last waste cycle.
Contact your facilities manager and/or EHS coordinator depending on your DLC.
Users are expected to post signage to indicate broken autoclaves and warn users.
Contact your EHS coordinator or biosafety officer to find alternative equipment or options.
Is there another way to validate/ensure an autoclave is working properly
The BSP team can provide data loggers to labs for autoclave validation. These data loggers are added to the autoclave’s regular cycle and continuously record temperature data. The collected information can be extracted and reviewed to ensure proper autoclave function. If you want to loan a data logger, please email BSP@mit.edu.
What supplies do I need?
Name
Catalog Number
Details
Source
Clear autoclave bag
14232-187
24X36 in(double thick)
VWR stockroom
Clear autoclave bag
1422-042
25X35 in
eCAT-VWR section
Clear autoclave bag
14220-044
36X45 in
eCAT-VWR section
Polypropylene tray
62662-048
eCAT-VWR section
Metal tray
19758-058
eCAT-VWR section
Temperature sensitive tape-lead free
1322-24MM
3M
Gloves rated for hot items
WLS40254-10
eCAT-VWR section
At MIT, autoclaves are used to sterilize waste and clean materials associated with biomedical research. The Autoclave Validation Program oversees compliance of biohazardous waste management at the state level (Massachusetts State Sanitary Code Chapter VIII 105 CMR 480.000) and assists in maintaining the institute’s international accreditation with AAALAC (Association for Assessment and Accreditation of Laboratory Animal Care). The program is managed by the EHS Biosafety Program, with local coordination and operational collaboration across the MIT campus.
The MIT Autoclave Validation Program performs challenge testing on all autoclaves used within MIT research facilities. Validation tests are performed with a small self-contained biological indicator (BI) containing a standard challenge population of Geobacillus stearothermophilus spores, a non-pathogenic thermophilic bacterium that is naturally resistant to high heat and pressure. Inactivation of G. stearothermophilus spores can be achieved with a 16-minute exposure to typical autoclave conditions of 121°C and 15psi, and as a result, these spores are an ideal biological indicator of microbial viability after autoclave sterilization.
The Autoclave Validation Program oversees compliance of biohazardous waste management and assists in maintaining the institute’s international accreditation with AAALAC (Association for Assessment and Accreditation of Laboratory Animal Care)
There are 4 cycles each year, and each autoclave on campus has to be validated. The validation process includes incubation of a Biological Indicator (BI) during a sterilization cycle, followed by overnight incubation of the BI at 65°C (either at the lab or by BSP staff)
Vials are available at the VWR room (56-070) and in the EHS office (N52-496). Vials can also be sent via interdepartmental mail. To request vials, please email BSP@mit.edu or contact your EHS coordinator.
The yearly validation cycle dates are:
Cycle number
Cycle dates
Cycle validation deadline
Q1
1/6 – 3/31
3/1
Q2
4/1 – 6/30
6/1
Q3
7/1 – 9/30
9/1
Q4
10/1 – 12/21
21/1
Autoclaved biological indicator (BI) may be dropped off in the VWR stockroom (56-070), returned to your BSP Officer, or returned directly to the EHS office (N52-495) in person or by mail. Returned vials will be processed by the BSP staff. Validators may complete the process in their own lab. See below for instructions.
Instructions for in-lab autoclave validation processing
Place the BI in the center of the load.
Select the cycle used for the sterilization of biohazardous waste. The cycle must include a minimum temperature of 121°C or 250°F for 30 minutes or longer, depending on size and compaction of the load. The full cycle time will take 60-90 minutes.
Remove the indicator when the cycle is complete and the load is cool enough to handle.
Process the autoclaved indicator and an indicator that has not been autoclaved (as a control) by first gently crushing the inner glass ampoule, this can be done by pressing on the outer plastic sheath near the brown cap; when broken you will see the purple media dampen the spore paper at the bottom of the vial. Then incubate at 55-60°C for 24 hours.
If growth occurs in the processed indicator, the autoclave has not inactivated the spores. Test the autoclave again with a new biological indicator and consider adding a secondary sterilization validation device like a chemical integrator. If, after a second attempt, the autoclave has not inactivated the spores, the autoclave must remain out of service until a successful biological indicator test is achieved. The person responsible must inform users not to use the autoclave and post a “Do Not Use” sign while the autoclave is out of service
Safety training on autoclave use
Please follow this autoclave use SOP to ensure the safe and effective operation of autoclaves in MIT research labs.
An online training on autoclave safety is available through the MIT Atlas Learning Center.
Go to the course catalog and search for “Autoclave Safety Training” (EHS00254w).
You also must complete an in-person training; this is typically done by your EHS Representative, experienced researcher, PI, or EHS coordinator.
Clean materials (glassware, Surgical tools, liquids, etc.) should be collected and placed in an autoclave-proof tray.
Run the appropriate cycle (121°C, 15 psi, 15 – 60 minutes).
After the run, allow the load to cool for at least 10 minutes.
Caution: Failure to allow sufficient cooling time may result in injury
Can I autoclave waste?
MIT disposes of biological/medical waste primarily using biowaste boxes,
Biological/medical waste is autoclaved under certain conditions (permit requirements, only effective method of waste inactivation, per specific approved risk assessment, etc.).
Please consult with your biosafety officer if you have questions or concerns.
How do I know if I can autoclave waste in a specific autoclave?
Autoclaves are designated and labeled “for waste.” Not all DLCs have autoclaves designated specifically for waste. If you are unsure whether you should autoclave your waste, please consult with your EHS coordinator or Biosafety contact.
Sterilization of waste
Follow the autoclave use SOP to ensure the safe and effective operation of the autoclave.
Waste should be collected and placed in a clear bag in a marked (biohazard labeled) transfer container.
For liquids, leave cap slightly OPEN for effective steam penetration during autoclaving and to prevent pressurization of the container and spills inside the autoclave.
Leave the bag OPEN for effective steam penetration during autoclaving.
Place waste bag in a secondary autoclave-proof container and run waste cycle (121°C, 15 psi, 60 minutes).
After the run, allow waste to cool for at least 10 minutes. Caution: The waste is hot. Wear appropriate PPE when handling hot waste
Failure to allow sufficient cooling time may result in injury.
Enter the waste run in the autoclave log binder, usually near the equipment.
Place treated and tagged waste bags into regular waste for custodial pickup.
Please consult your EHS coordinator or BSP@mit.edu for questions and concerns.
Autoclaves that are designated for waste must:
Autoclaves designated for waste must:
Be labeled “For Waste” on or near the autoclave. (Note: Not all DLCs have autoclaves specifically designated for waste. If you are unsure whether to autoclave your waste, please consult your EHS coordinator or Biosafety officer.)
Be updated in the QuickBase database as a waste autoclave. Please confirm with your EHS coordinator or Biosafety contact whether your autoclave is designated for waste.
Have a completed Autoclave Waste Log template placed nearby. This log must be retained for three years after the last waste cycle.
Contact your facilities manager and/or EHS coordinator depending on your DLC.
Users are expected to post signage to indicate broken autoclaves and warn users.
Contact your EHS coordinator or biosafety officer to find alternative equipment or options.
Is there another way to validate/ensure an autoclave is working properly
The BSP team can provide data loggers to labs for autoclave validation. These data loggers are added to the autoclave’s regular cycle and continuously record temperature data. The collected information can be extracted and reviewed to ensure proper autoclave function. If you want to loan a data logger, please email BSP@mit.edu.
