Select Agent Toxins

Principal Investigators must be registered to work with a biological toxin. Registration is typically done through an amendment to the PI’s Biological Research Registration. This process allows EHS Biosafety and the Committee on the Assessment of Biohazards (CAB-ESCRO) to review the proposed research and evaluate any safety and regulatory concerns. Please see our Registration webpage for more details on this process.

Prior to working with a Select Agent toxin, each researcher must complete Use of Biological Toxins Training (EHS00215C). This training is given upon request. Please contact BSP@mit.edu for more information.

The lab must be registered to work with toxins prior to ordering.
Toxins can’t be purchased with a P. Card.

Please follow the steps below:

• Order through Buy2pay
• Attach a copy of the last page of the most up to date toxin inventory log
• Under Commodity choose  Biological – Select Toxins
• BSP will review all orders on Buy2pay website and will give final approval
• Orders are delivered directly to the lab.
• Toxins are placed in lockbox(es)
• Toxins are added to toxin inventory
• New inventory log is emailed to the Biosafety Office. Email your direct Biosafety officer or email BSP@mit.edu, with the new toxin amount and date.

Please contact your biosafety officer for more details on how to order a Select Agent Toxin, or Email us at BSP@mit.edu

Authorized and EHS trained laboratory personnel should be mentored and supervised by a more experienced researcher. All researchers working with toxins must be familiar with the laboratory written protocol and/or Standard Operating Procedure (SOP) detailing the technique/experiments steps/handling, containment equipment, personal protective equipment (PPE), storage, disposal, internal communication, and emergency response. This protocol or SOP should be updated as needed and kept accessible to users. Contact BSP at  BSP@mit.edu for a template of Toxin SOP.

The following records should be maintained at the laboratory’s location:

• An up to date toxin inventory
• Copies of each toxin purchase order

Records are kept by Principal Investigator for at least 3 years after the select toxin is completely used up or is destroyed

Storage of select toxins shall comply with the following criteria:

• The stock solution should be secured in a lockbox. Contact BSP@mit.edu if you do not have one in the lab.
• Tubes, bottles, aliquots should be clearly labeled and numbered for rapid identification of the material. Information should include, volume, date of preparation, vial # and user/prepared name.
• Tubes should be, if possible, rubber septum with screw or crimped caps for added safety and should be wrapped with parafilm and kept in a spill proof secondary container e.g. properly labeled zipper storage bag or box.
• The inventory shall be accurate and up to date.
• Access should only be given to properly trained personnel

Security

Custody of select agent toxins is the responsibility of the PI once EHS-BSP delivers the select agent toxins to the approved and trained laboratory personnel. All stock solutions of select agent toxins are kept under lock and key. Diluted working solutions may be stored outside of the lock box but the amounts of diluted solutions should be minimized.

Investigators shall only dispose of toxins using EHS BSP approved methods. All generated select toxin liquid waste shall be treated as hazardous chemical waste and labeled with a “red tag” as “inactivated toxin” with appropriate required information before offering it for pick up by EHS. Highly diluted toxin liquid waste can be poured down the drain after properly inactivated by an EHS BSP approved method.

Solid and sharp wastes shall be assessed to determine level of toxin residue before disposing  in the proper waste stream (e.g. biowaste box, puncture proof sharp container, or as chemical hazardous waste where appropriate). If necessary, researchers are encouraged to contact EHS-BSP at BSP@mit.edu for assistance with this assessment.

Table 1. Chemical Inactivation of toxins

Allow at least 30 minutes of contact time for the inactivation of the toxin

Chemical Abbreviations: NaOCl = Sodium hypochlorite   NaOH = Sodium hydroxide

Table 2. Autoclave heat Inactivation of toxins

121°C for 1 hour, liquid cycle, slow exhaust

Recommendations from references on the tables:

(1) For complete inactivation of T-2 mycotoxins, all liquid samples, accidental spills, and non-burnable waste be soaked in 2.5% sodium hypochlorite with 0.25N sodium hydroxide for 4 hours.1
(2) Inactivation for Saxitoxin, Tetrodotoxin, Ricin, Botulinum toxin, or Staphylococcal Enterotoxins B (SEB), exposure of 30 minutes to 1% sodium hypochlorite is an effective procedure for working solutions, equipment, working area and spills.1
(3) Autoclaving can be used for protein toxins, i.e. Ricin, Botulinum toxin, and Staphylococcus Enterotoxin B (SEB), but should not be used with any of the low molecular toxins.

Select Agent Toxin Inactivation and Destruction Table

PIs shall not allow purchase of select toxins for stock piling, distribution, transfer or sharing. PIs are expected to acquire select toxins ONLY through B2P, use all of the select toxins under their custody, and have complete control of their inventory.

EHS-BSP shall assess special circumstances on a case by case basis upon PI request. Federal agencies retain the authority to, without prior notification, inspect and copy or request the submission of documentation.

Shipping and receiving

MIT Laboratories are not certified or authorized to ship via carrier (e.g. FedEx), courier, hand-carried or public transportation without assistance from EHS certified shippers; or receive select toxins under any circumstances without EHS-BSP Office involvement.

Upon discovery of the theft or loss of select toxins, laboratory personnel must immediately notify the PI and EHS-BSP 617-452-3477 /BSP@mit.edu. EHS-BSP Deputy Director and SAT program manager shall notify appropriate agencies. Thefts or losses must be reported even if the select toxin is subsequently recovered or the responsible parties are identified.

If at all possible, order toxins in liquid form rather than as a powder. Powders present the greatest risk during preparation of stock solutions due to possible aerosolization and danger of inhalation during handling. If the toxin is only available in powder form, order the toxin in vials that provide safety features (see table below). Avoid ordering toxins in powder form in scaled glass ampoules, as scaled glass ampoules are the most difficult to handle safely.

Some of the most commonly used vendors sell toxins in screw cap vials. These must also be handled carefully making sure the powder remains pelleted at the bottom of the tube. This can be accomplished by quickly centrifuging the vial.

Steps to generate stock solution from powders or liquids:

Put on personal protective equipment (PPE):

• Gloves: Gloves shouldn’t be permeable to toxin or solvent; double gloving is preferable
• Lab coat
• Safety goggles to protect from splash and/or impact

When handling concentrated toxin, work inside a containment device such as a chemical fume hood or Biosafety Cabinet (BSC):

• Remove any unneeded chemicals, satellite accumulation areas (SAAs), or unnecessary equipment (as reasonable)
• Line the work surface of the chemical fume hood or BSC with a plastic backed absorbent liner

If the toxin comes in regular screw cap vials:

• Quickly centrifuge the tube to pellet the powder
• Place the vial into a rack or tube holder in the chemical fume hood or BSC
• Open the screw cap vial carefully

If the toxin comes with a rubber septum (including screw caps):
Do not remove the rubber septum

• Use a blunt-tipped needle or pipet tip to inject a measured amount of buffer/solvent through the rubber membrane
• Cover rubber septum with parafilm and mix carefully to avoid aerosol generation
• Briefly spin in a centrifuge to bring the liquid on the inside of the cap or near the lid

If the toxin comes in a pre-weighed amount in a glass ampoule:

• Nick the ampoule with a file or file at the narrow end of the neck
• Wrap the ampoule in disinfectant-wetted cotton or paper towel
• Hold the ampoule away from you
• Snap the ampoule to open at the nick

For other containers:

• Inject measured amount of buffer/solvent into the vial/ampoule with a pipette tip or blunt-tipped needle
• Dissolve the powder and mix liquid contents carefully to avoid aerosol generation
• Mix to give a known concentration stock solution.

Stock solution division

• Use a pipette tip to remove contents
• Divide the solution into multiple tubes to reduce repeated freeze-thaw cycles
• These are the stock solutions, with a known amount of toxin in a known volume

Enter the volume and number of tubes of the stock solution in a new toxin inventory.

Store all stock solutions in a secured lockbox.

Waste handling:

• Treat the glass vial, ampoule or screw cap vial with an appropriate concentration of disinfectant (see your “Toxin Use SOP”) for the appropriate contact time.
• Dispose of glass vial(s), ampoule(s), needles, syringes in a puncture-proof sharp container
• Once full, these containers are be capped tightly/sealed and placed inside a biowaste box for disposal

Wipe down chemical fume hood or BSC surface with an appropriate concentration of disinfectant.

Collect all solid waste (gloves, plastic-backed absorbent paper) in a plastic bag and dispose of in biowaste box

If contamination is significant (such as in the case of a spill of concentrated toxin), this waste should be segregated as chemical hazardous waste. Please contact BSP if this situation arises for further guidance and coordination on the proper disposal. Under no circumstance must any select toxin waste reach public sewage systems.