Permissible Select Toxin Amounts and Biologically Derived Toxins with LD50 ≤ 100µg/kg

Introduction

This document establishes guidelines and policies for the possession, use, and disposal of select agent toxins quantities that are exempt from registration requirements of the Select Agent regulations and biological toxins with a median lethal dose (LD50) of ≤ 100 µg/kg. Select Agent Toxins are biological agents or toxins deemed threatening to the public, animal or plant health, or animal and plant products. A laboratory facility that possesses more than the amount specified for each toxin must register with the CDC/USDA and fulfill all federal requirements in (1) 42 CFR Part 73, Final Rule for Possession, Use, and Transfer of Select Agents and Toxins, (2) 7 CFR Part 331, and (3) 9 CFR Part 121, USDA Final Rule for Possession, Use, and Transfer of Biological Agents and Toxins.1 Laboratory having less than the stated amount are exempt from most federal select agent requirements. However, they must still adhere to 42 CFR Part 73.3 Section 16 regarding due diligence and the policies outlined in this document.2 Due diligence encompasses a set of practices that would be executed by a reasonable person so as not to cause harm to other persons, the environment, or animals.2 The Federal Select Agent Program should be consulted for further information. 

Back to Top

Scope and Requirements

The procedures outlined in this document apply to all laboratories at Cornell University working with permissible amounts of select toxins and biologically derived toxins with LD50 ≤100 µg/kg. The purpose of this document is to ensure federal compliance when storing, handling, or transferring select toxins as described in:  

• 42 CFR Part 73, Final Rule for Possession, Use, and Transfer of Select Agents and Toxins

• 7 CFR Part 331, Possession, Use, And Transfer of Select Agents And Toxins

• 9 CFR Part 121, USDA Final Rule for Possession, Use, and Transfer of Biological Agents and Toxins (Select Agents Regulations webpage)1

• Title 6 NYCRR Parts 360-366 and 369, Solid Waste Management

Back to Top

Roles and Responsibilities

Biological Safety Office

It is the responsibility of the Biological Safety Officer and delegates to identify and communicate legal, regulatory, and policy requirements associated with the program.

Principal Investigator

The Principal Investigator’s (PI) responsibilities include:

• Complying with all state, federal and local laws regarding the toxin possessed/in use

• Register materials with Institutional Biosafety Committee, complete Biotoxin Standard Operating Procedures, and attach them to the IBC Memorandum of Understanding and Agreement (MUA) for review.

• Train all personnel, students, and postdocs on the appropriate use and recordkeeping for toxins used within the PI’s research area(s). Required records include:

  • Inventory, including information on toxins, and amounts received, transferred, or destroyed.

  • Maintain records of training.

• For Select Toxins Only: Perform due diligence regarding all transfers of Select Agent Toxin from the Principal Investigator’s laboratory – whether on campus or outside of Cornell University. The definition and requirements for Due Diligence regarding Select Agent Toxins are in 42 CFR Part 73.3 Section 16. A Frequently Asked Questions (FAQ) document is available through the federal select agent program website. Please note that compliance with practices of Due Diligence is required for any quantity of Select Agent Toxin.2

  • Report to the federal select agent program if the transferor detects a known or suspected violation of Federal law or becomes aware of suspicious activity related to the shipped select toxin. If reporting, include pertinent information of the person requesting and using the select toxins (e.g., name, institution name, address, telephone number, and e-mail address). A transferor can contact FSAP either by e-mailing CDC: LRSAT@cdc.gov or APHIS: DASAT@usda.gov or by calling FSAP (CDC: 404-718-2000 or APHIS: 301-851-2070).2

Research Area Personnel

• Responsible for completion of all training relevant to work with select agent toxins offered by the University through EHS programs and their laboratories.

• Responsible for using toxins in a manner consistent with active research in their laboratories as well as maintenance of accurate records of their use during research.

Back to Top

Procedures

Registration with Institutional Biosafety Committee

The laboratory will register ownership and use of the toxin(s) with the Institutional Biosafety Committee (IBC) before purchase or transfer and start of work. Please contact the IBC through the Office of Research Integrity and Assurance with any questions: cu_ibc@cornell.edu or 607-255-7219.

Back to Top

Training

The Principal Investigator (PI) or lab supervisor ensures that personnel understand the hazards associated with a toxin, and the appropriate controls and precautions to take in the event of an emergency. The experimental protocol, including all necessary precautions, is reviewed with all research personnel and students. Appropriate handling and knowledge of laboratory-specific procedures are the responsibility of the Principal Investigator.

EHS required training include:

• EHS 2655 – Cornell University Health and Safety Basics

• EHS 2555 - Laboratory Safety before beginning work with Select Agent Toxins. This course is a general laboratory safety course familiarizing lab personnel with safety procedures in the Cornell University laboratory environment.

• EHS 2716 – Chemical Waste Disposal. This course reviews the disposal of hazardous substances.

• EHS 2394 – Cleaning Up Small Lab Spills

EHS recommended training includes:

• CVM 1074 – Working Safely in Biosafety Cabinet and other Hoods

Back to Top

Storage and Security

PI or lab supervisor maintains the security of the areas containing select toxins and/or biologically derived toxins with LD50 ≤100 µg/kg. Security requirements are listed below.

1. Secure the toxins under lock and key. Lockboxes are securely affixed to surfaces or larger equipment (full-sized refrigerators, freezers) rendering removal from laboratories difficult or impossible. A lockbox used for the storage of controlled substances may be suitable for the storage of the toxin.

2. Restrict access to toxins to those authorized to perform work in the area. Restrict access of unauthorized personnel.

Back to Top

Laboratory Handling

• Wear appropriate PPE including, at minimum, eye protection, laboratory coat, and gloves. Respiratory protection and a face shield may also be recommended. Avoid contact with eyes, skin, and clothing. Cover all breaks in the skin (including chapped skin).

• If possible, order select toxins in liquid form rather than powder form. If only a powder form of the toxin is available, the toxin is ordered in vials with a rubber septum and crimped metal caps. 

• Any area where toxins are manipulated is labeled as an OSHA “Designated Area.” This includes all chemical fume hoods, benchtops, or the entire laboratory. 

• The lab where the material is being handled must have an approved and certified emergency eyewash and safety shower. 

• Consider class II, type B2 biosafety cabinets, or chemical fume hoods for all work requiring the manipulation of dry powder that has the potential for aerosolization. Sealed vials containing dry powder are reconstituted with an appropriate liquid under original sealed conditions. Concentrated stock solutions of the toxins may be used in a chemical fume hood. Consider the filtration of exhaust, location of exhaust, and concentration of potential discharge. Avoid the use of ductless hoods or ductless biosafety cabinets. 

• Wash hands thoroughly after handling, and avoid prolonged or repeated exposure to, toxins. 

Back to Top

Waste Handling and Disposal

Per New York State law, before disposal, destroy Select Agent Biological Toxins by using appropriate chemical or physical agents (i.e., sodium hypochlorite and/or sodium hydroxide solution or by autoclave). Table 1 demonstrates inactivation methods (Chemical and Physical) for select toxins. The following procedures are recommended when destroying toxins: 

1. Inform EHS through e-mailing askEHS@cornell.edu or call 607-255-8200.  

2. Use appropriate personal protective equipment.  

3. Perform the work inside a chemical fume hood. Ensure that the fume hood sash is at the appropriate working height.  

4. When chemical inactivation is appropriate, prepare a fresh solution of the chemical agent as shown in Table 1.  

5. Line the work area with plastic-backed absorbent pads.  

6. Toxins in powder form contained in vials sealed with a rubber septum are dissolved in solution, through the septum, without opening the container.  

7. For a toxin already in solution, open the container and place it in a second plastic container with a sealable lid.  

8. Carefully add the appropriate amount of the chemical deactivating agent to the toxin.  

9. Allow at least 30 minutes for the chemical agent to inactivate the toxin.  

10. Replace the cap on the primary toxin container.  

11. Place and secure the lid on the secondary container.  

12. Label and dispose of the container as hazardous chemical waste. 

Back to Top

Inactivation/Decontamination

Inactivation procedures for the toxins can be found in Table 1. 

• Exposure for 30 minutes to 1.0% sodium hypochlorite solution is an effective procedure for laboratory (utensils, glassware, working solutions, instruments, animal cages, working area, and equipment) inactivation of saxitoxin, tetrodotoxin, ricin, botulinum toxin, or staphylococcal enterotoxins (SEB).3 

• For complete inactivation of T2 mycotoxin, all liquid samples, accidental spills, and non-burnable waste it is recommended that all materials be soaked in 2.5% sodium hypochlorite with 0.25N sodium hydroxide for 4 hours; expose cages and bedding from animals exposed to T-2 mycotoxin to 0.25% sodium hypochlorite and 0.25N sodium hydroxide for 4 hours.3

Back to Top

Disposal

Disposal of Liquid waste

The residual toxin in the solution is inactivated by the laboratory before disposal. Inactivated toxin effluent is placed within a clearly labeled closed container in a chemical fume hood and disposed of as hazardous chemical waste.  

Disposal of Solid waste 

All solid waste must be transferred to the College of Veterinary Medicine Waste Management Facility for final disposition.

For heat-labile toxins that can be inactivated by autoclaving (see Table 1 or consult EHS) 

Discard used gloves, paper towels, and other disposable supplies, containing the residual toxin, in a red biohazard bag. Seal and place the biohazard bag in a regulated medical waste container. Used needles, pipet tips, and other sharps are discarded in a red biohazard sharps container.  

For heat-stable toxins that require chemical inactivation/incineration (see Table 1 or consult EHS)  

Discard used gloves, paper towels, and other disposable supplies, containing the residual toxin, in a yellow biohazard bag. Seal and place the biohazard bag in a regulated medical waste container. Used needles, pipet tips, and other sharps are discarded in a yellow biohazard sharps container. 

Disposal of any Select Agent must follow the final rules on Select Agents and Toxins.

Table 1. Chemical and Physical Inactivation of Toxins3

*The minimal effective concentration of NaOCl was dependent on toxin and contact time; all LMW toxins tested were inactivated at least 99% by treatment with 2.5% NaOCl or with a combination of 0.25% NaOCl and 0.25 N NaOH.

Back to Top

Recordkeeping, Reporting, and Monitoring

The following records are maintained at the laboratory’s location:

• Maintain an accurate and current inventory (Provided Below) of each toxin in your possession and a list of the individuals approved for access to the toxins.

  • Destruction of the toxins is properly documented and recorded by the laboratory. The following information is retained by the laboratory upon the destruction of the toxin and kept in the PI’s records for five years.

    • Name of toxin

    • Copy of SDS & Manufacturer information, if available

    • Quantity

    • The procedure followed for destruction

    • Name of the person who destroyed the toxin

    • Date and time of destruction

    • Location of destruction

• Record of laboratory-specific training.

• Copy of the laboratory-specific SOP (Docx)

• Select Toxins Only: Due diligence record following any transfers.

Back to Top