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.
Autoclaved biological indicator (BI) may be dropped off in the 56-070, returned to your BSP Officer, or directly to the EHS office 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
Personal protective equipment (PPE):
safety glasses
long sleeves and a lab coat
Gloves rated for hot items
Sterilization of clean materials
Clean materials (wrapped, liquids, etc.) should be collected and placed in an autoclave-proof tray.
Run the appropriate cycle (121°C, 15 psi, at least 15 minutes).
After the run, allow 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 for waste. If you are not sure if you should autoclave your waste, please consult with your EHS coordinator or Biosafety contact.
Sterilization of waste
Waste should be collected and placed in clear bag in marked (biohazard labeled) floor 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 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 waste run in 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.
Personal protective equipment (PPE):
Safety glasses/goggles/face shield
A lab coat
Gloves rated for hot items (can wear nitrile gloves below)
Autoclaves that are designated for waste have to:
Be labeled “for waste”(not all DLCs have autoclaves designated for waste, if you are not sure if you should autoclave your waste, please consult with your Biosafety contact)
Be updated on the QuickBase database as a waste autoclave. Please check with your EHS coordinator or Biosafety officer if your autoclave is designated for waste.
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.
Autoclaved biological indicator (BI) may be dropped off in the 56-070, returned to your BSP Officer, or directly to the EHS office 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
Personal protective equipment (PPE):
safety glasses
long sleeves and a lab coat
Gloves rated for hot items
Sterilization of clean materials
Clean materials (wrapped, liquids, etc.) should be collected and placed in an autoclave-proof tray.
Run the appropriate cycle (121°C, 15 psi, at least 15 minutes).
After the run, allow 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 for waste. If you are not sure if you should autoclave your waste, please consult with your EHS coordinator or Biosafety contact.
Sterilization of waste
Waste should be collected and placed in clear bag in marked (biohazard labeled) floor 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 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 waste run in 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.
Personal protective equipment (PPE):
Safety glasses/goggles/face shield
A lab coat
Gloves rated for hot items (can wear nitrile gloves below)
Autoclaves that are designated for waste have to:
Be labeled “for waste”(not all DLCs have autoclaves designated for waste, if you are not sure if you should autoclave your waste, please consult with your Biosafety contact)
Be updated on the QuickBase database as a waste autoclave. Please check with your EHS coordinator or Biosafety officer if your autoclave is designated for waste.
