Principal investigators (PIs), Laboratory Safety Officers (LSOs), and laboratory personnel are responsible for properly identifying and decontaminating regulated medical waste, including potentially infectious material and recombinant DNA (rDNA) or synthetic Nucleic Acid (sNA) waste, before disposal. Procedures to identify, package, and transport biohazardous waste are provided below. Refer to the poster guidance, Cell/Tissue Culture Waste Disposal Guide for Laboratories, for additional guidance.

Solid waste potentially contaminated with infectious agents or rDNA/sNA designated for BSL2 (or higher) containment require decontamination before disposal. Decontamination of solid waste can be achieved either via chemical disinfection or steam sterilization (i.e., autoclaving).

Due to regulatory requirements, biological waste must be managed as Regulated Medical Waste, which includes biohazardous waste decontaminated by laboratory staff. RMW can only be disposed of as regular trash after it has undergone treatment at a licensed facility. Stericycle is the approved Mount Sinai vendor for the compliant collection, transportation, treatment and disposal of biohazardous wastes. Our institutional policy requires decontamination for specific material before disposal in order to mitigate potential hazards when it’s handled by Building Services and transported offsite. It also helps ensure compliance with best practices, standards, and regulations related to biosafety and transportation safety.

If you have access to an autoclave and have completed the CITI training course, Autoclave Safety Training, then you can decontaminate and dispose of biohazardous waste yourself. Steam sterilization with an autoclave effectively inactivates infections microorganisms. All research and clinical laboratories must comply with New York State regulations that apply to the maintenance and use of steam autoclaves that are used to decontaminate biohazardous waste.

Regulated Medical Waste

Mount Sinai considers the following biohazardous waste materials as RMW:

• Sharps waste (e.g., needles, syringes, broken glassware)
• Human and nonhuman primate blood, tissue, body fluids, and cell lines
• Cultures or stocks of biohazardous agents (e.g., bacteria, rickettsia, fungi, viruses, protozoa, parasites, and prions)
• Recombinant DNA (rDNA) or synthetic Nucleic Acids (sNA), which includes waste products from laboratory procedures involving plasmids, viral vectors, Escherichia coli (E. coli), yeasts, and naked nucleic acids.
• Laboratory waste items that are potentially infectious materials (i.e., used PPE, culture dishes, tubes, etc., that have come into contact with a biohazard)
• Laboratory animal waste (e.g. bedding, carcasses, and body parts) that have been exposed to rDNA or any biohazardous agent
• Human pathological waste

The New York State Department of Environmental Conservation defines RMW as material generated in biomedical research, production and testing of biologicals or health care, including:

• Infectious animal waste
• Human pathological waste
• Human blood and blood products
• Needles and syringes (sharps)
• Cultures and stocks (microbiological materials)
• Other infectious waste (e.g. materials contaminated with infectious agents such as the hemorrhagic fever viruses

Disposal of Solid Biohazardous Waste

• Collect solid biohazardous waste in autoclave-safe biohazard waste bags. Biohazardous waste that is known to be infectious should be double-bagged with autoclave-safe biohazard waste bags
• Add water to the autoclave-safe biohazard waste bags before closure
• Loosely tie the autoclave-safe biohazard waste bags before autoclaving to allow steam to escape
• Secure the bag inside a rigid, leak-proof container (e.g., low-density polyethylene (LDPE) or stainless steel tray) that allows the bag’s biohazard symbol to be visible

Disposal of Intact Glass and Plastic Waste

• Collect glass and plastic waste in a puncture-resistant container (e.g., Stericycle red sharps bin or a sturdy cardboard box lined with a red Regulated Medical Waste bag)
• Laboratory waste items that are potentially contaminated with infectious materials or recombinant/synthetic nucleic acid molecules designated for BS2 containment should be decontaminated prior to disposal (e.g., by steam autoclave sterilization)

Disposal of Biohazardous Sharps Waste

• Collect sharps (e.g., broken glassware or pipet tips) in a puncture-resistant container (e.g., Stericycle red sharps container)
• Do not mix with any other type of waste

Liquid Biohazardous Waste Disposal

• Liquid biohazardous waste must be decontaminated before disposal down the laboratory sink into the sanitary sewer system
• Collect liquid biohazardous waste in leak-proof, rigid containers labeled with a biohazard symbol
• Liquid biohazardous waste must be decontaminated with a chemical disinfectant following manufacturer’s instructions or by steam autoclave sterilization
• Sodium hypochlorite is generally the standard chemical disinfectant for tissue culture waste disposal since is acceptable for drain disposal. If another chemical disinfectant is used, then the chemical disinfectant would need to be evaluated by EnvH&S to determine if it is acceptable for drain disposal per EPA regulations and/or other local and state standards for sewage discharge (e.g., NYC Department of Environmental Protection)
• If sodium hypochlorite (i.e., bleach) is used as a disinfectant, then add hypochlorite solution to equal a final concentration of 10 percent. Allow an eight-hour contact time before disposal down the laboratory sink.

Human Clinical Pathology Waste and Nonhuman Primate Carcasses

Incineration is required for human clinical pathology waste and nonhuman primate carcasses. Make disposal arrangements before obtaining human or nonhuman primate pathological samples. Do not dispose of clinical pathology waste with other biohazardous wastes.

Laboratory Animal Waste

Incineration is required for recognizable tissue, body parts, and carcasses from vertebrate laboratory animals. The Center for Comparative Medicine and Surgery (CCMS) oversees the disposal of these types of laboratory animal wastes.

Transporting Biohazardous Waste

If transporting biohazardous waste from the point of collection to a centralized decontamination center, then secure the biohazardous waste in leak-proof secondary containers, which should be marked with the universal biohazard symbol.

Regulated Medical Waste Disposal

All Regulated Medical Waste must be discarded into Stericycle red bins or bagged in red Regulated Medical Waste bags that are preprinted with the Mount Sinai institutional address (see Figure 1). To request pickup of Regulated Medical Waste or request a supply of red Regulated Medical Waste bags, send an email to Building Services. Your email should indicate:

• Name of the person responsible for the RMW
• The location (building and room number)
• Amount of RMW to be picked up, and/or
• Amount of red RMW bags needed

Biohazardous/Sharps Waste Containers

To add additional Sharps Containers to an existing location, please contact Joseph G. Marinello and Jhonier Hernandez, who will create an order for delivery of additional red biohazardous waste containers. Please allow 7 to 14 days for delivery depending on the quantity of containers requested.

To add on a new laboratory or location that does not have sharp containers, please contact Joseph G. Marinello and Reshma Smith and provide the following information:

• Name of the Lab
• Building Name
• Floor
• Laboratory Room number and Name of Room (Tissue Culture, Micro etc.)
• Size of sharp container (8- or 17-gallon) requested and quantity needed for each size

If your laboratory will move to a new location on campus, then please contact Joseph G. Marinello and Reshma Smith, who will modify your existing account to reflect the change in location.

Autoclaving is the most effective and reliable means of sterilizing laboratory materials. The steam sterilizer (autoclave) is a steam-pressure vessel, which must be operated in a proper manner to ensure inactivation of microbial agents and the safety of the operator. A steam autoclave is a gravity flow sterilizer, which must reach a minimum temperature of 121°C (250°F) at 15 pounds per square inch (psi) of internal steam pressure. Depending on the volume and density, the biohazardous material may require 60-120 minutes for sterilization.

DSOs/LSOs must provide training to research personnel on the correct preparation of biohazardous materials for sterilization, and on the correct use of steam autoclaves. Autoclave users should understand the time, temperature, and pressure relationships required for proper decontamination of biohazardous materials. Users should complete the CITI training course, Autoclave Safety Training, prior to using an autoclave.

Personnel should wear proper personal protective equipment when operating the autoclave, particularly when unloading the autoclave, including:

• Lab coat
• Heat resistant gloves
• Closed-toe shoes
• Eye protection

Steam autoclave sterilizers should be certified during annual preventative maintenance. Autoclave door clamps and seals should be inspected for wear and damage over time. Prior to using a steam autoclave, inspect the autoclave components for proper operation and remove any debris from the strainer for the autoclave chamber floor drain.

After the steam autoclave completes a sterilization cycle, check the printout to confirm the cycle reached the correct temperature and pressure for the configured exposure time. If the printout indicates an aborted autoclave cycle, then assume that the contents are still potentially biohazardous.

Steam Sterilization Process Validation

It really is important when autoclaving waste to have validated the process with biological indicators included inside surrogate loads that represent normal biohazardous waste (i.e., number, content, and position of autoclave-safe biohazard bags). Biological indicator systems based on Geobacillus stearothermophilus spores are designed to demonstrate that the programmed steam autoclave cycle effectively inactivates microorganisms. These systems assure that the right combination of Time, Temperature, and Steam Saturation occurred during the cycle to ensure sterilization.

Steam Sterilization Process Controls

Heat-sensitive autoclave tape is a type of chemical indicator that only confirms that the steam sterilizer reached the minimum processing temperature. A change in color of heat-sensitive tape does not indicate that the steam autoclave cycle has decontaminated the biohazardous materials.

The Biosafety Program recommends the use of 3M™ Comply™ Thermalog™ or 3M™ Comply™ SteriGage™ chemical integrators to confirm the sterilization process. Chemical integrators provide immediate, easy to read “Accept” or “Reject” sterilization results. You can purchase these chemical integrators from any major commercial laboratory supply vendor.

Consideration for Safe and Effective Autoclave Use 

• Be sure the autoclave is OFF, and the pressure is regulated before opening the doors. Autoclave doors should be opened slowly, keeping the head, face, and hands away from the opening to prevent steam contact 

• Check the autoclave for items left behind before loading 

• Waste should be autoclaved in specific autoclaving bags (that have the biohazard symbol) that are designed to withstand the extreme conditions of the autoclaving process. A chemical integrator should be taped to the outside of the bag being autoclaved (one integrator per load). Bags must be placed into a secondary, melt-proof container for autoclaving 

• Load each autoclave in accordance with the manufacturer’s recommendations to ensure complete sterilization. DO NOT OVERLOAD THE AUTOCLAVE 

• If you are placing bottles in the autoclave, be sure to loosen caps so pressure buildup doesn't cause bottles to shatter or rupture during the run 

• Do not overload the autoclave bag. The autoclave steam and heat cannot penetrate the interior of an overloaded bag. The outer contents of the bag will be sterilized, but the inner part of the bag will essentially be unaffected by the autoclave cycle 

• Since steam has to be in contact with the material being sterilized to be effective, bags and containers must be sealed loosely to allow steam penetration into the bag 

• Do not mix autoclave bags and other items to be autoclaved in the same autoclave cycle. Liquid media requires a shorter cycle, while autoclavable medical waste requires more time to be effectively sterilized 

• The Autoclave door should be locked before operation. Be sure not to over-tighten the door to prevent damaging the seal. Most autoclaves will not operate if the door is not completely closed and locked. 

• Use caution when opening the autoclave door. Allow superheated steam to exit 

• Use caution when handling a bag in case sharp objects have been inadvertently placed in the bag. Never lift a bag from the bottom of the bag to load it into the chamber. Handle the bag from the top. 

• For non-liquid loads, allow 15 minutes for cooling prior to touching with bare hands 

• For liquid loads, allow to stand at least one hour before touching with bare hands 

• Watch out for pressurized containers. Superheated liquids may spurt from sealed containers. Never seal a container of liquid with a cork that may cause a pressurized explosion inside the autoclave 

• Agar plates will melt, and the agar will become liquefied. Avoid coming in contact with this molten liquid. Use a secondary autoclavable tray to catch any potential leakage from the bag that would otherwise leak into the autoclave 

• Glassware may crack or shatter if cold liquid comes in contact with this superheated glassware. If glass breaks in the autoclave, use tongs, forceps, or other mechanical means to recover the fragments; make sure that the autoclave has been cooled down to avoid surface burns 

• Use an absorbent liner for glass vessels containing liquid. Never put autoclave bags or glassware directly in contact with the bottom of the autoclave 

• Do not store combustible materials near autoclaves 
• Use a tray with a solid bottom and sides to contain the material being autoclaved to catch any spills, should they occur. Before placing plastic materials into autoclaves, check to ensure they are compatible. All manufacturer's safety precautions should be observed in addition to these listed in this document 

Compatible/Incompatible Material 

Never autoclave: 

• Flammable, reactive, corrosive, toxic, or radioactive materials 

• Household bleach 

• Any liquid in a sealed container. 

• Any material contained in such a manner that it touches the interior surfaces of the autoclave. 

• Paraffin-embedded tissue. 
• Glass 
• Only Pyrex^® or Type I borosilicate glass is autoclavable. When autoclaving liquids in Pyrex^® containers, do not fill more than two-thirds full and do not seal the container. 
• Plastic 
• Polypropylene is an inexpensive resin that can resist autoclave temperatures. Polypropylene containers are often used as secondary containers to hold materials that are autoclaved. Polycarbonate can also withstand high temperatures. Polystyrene (PS), polyvinyl chloride (PVC), nylon, acrylic, low-density polyethylene (LDPE), and high-density polyethylene (HDPE) lab ware and polyurethane tubing are not autoclavable under any conditions. 
• Gloves 
• Gloves must be placed inside an autoclavable biohazard bag and exposed to a steam setting; gloves will melt slightly but will not burn when autoclaved in this manner. 
•  Stainless steel 
• Most metals are designed for extreme conditions and are intended to be sterilized. Make sure to remove any plastics, liners, and other items that may melt or combust. 
• Paper 
• Paper is combustible and should not be placed directly inside an autoclave. It should be autoclaved in a waste bag in a biobag setting to prevent fire. 
• Pipette tips 
• Most pipette tips are autoclavable. Some of these tips are plastic; some are high-density polyethylene. In general, pipette tips should only enter the autoclave as waste inside of an approved biohazards bag and should always be sterilized in a steam-producing setting. 

As part of the commitment of the Icahn School of Medicine at Mount Sinai to establish and maintain a “Culture of Safety”, the Occupational Health and Safety Program (OHSP) is designed to prevent the occurrence of injuries and occupational diseases. Workforce Health and Environmental Health and Safety (EnvH&S) implement elements of the institution’s OHSP.

Medical Surveillance Program

Workforce Health provides services to all employees within the Mount Sinai Health System and the Icahn School of Medicine at Mount Sinai. Workforce Health offers evaluation and treatment for work-related illnesses and injuries, performs return-to-work evaluations, administers the Workplace Accommodations Program, and administers medical surveillance programs, including testing and immunizations mandated by OSHA, New York State, New York City, and Mount Sinai.

Medical surveillance testing may be required for Researchers who directly handle biohazardous materials or for laboratory visitors who are potentially exposed to biohazardous materials. Depending on the biohazardous material and the scope of research, medical surveillance testing may be required. The IBC, Biosafety, and Workforce Health will determine optimal surveillance testing protocols. Each laboratory member must complete an Annual Health Assessment, which is reviewed by Workforce Health to: i) assess personnel health fitness to work in the laboratory environment, ii) ensure that vaccination status against bloodborne pathogens is up-to-date, and iii) provide individual counseling as needed.

Workforce Health requires employees to schedule appointments for completing occupational health compliance (i.e., influenza immunization,  Purified Protein Derivatives/QuantiFERON-TB Gold Tests/Tuberculosis screening), Annual Health Assessments, return to work clearances, and work-related injuries and follow-ups. Each employee should email his/her completed Annual Health Assessment to Workforce Health.

Respiratory Protection Program

Environmental Health and Safety (EnvH&S) maintains programs related to compliance with federal, state, and local regulations involving worker health and safety issues, as well as exposure of the public, patients, and visitors to safety hazards. The Occupational Safety and Health Administration’s (OSHA) Standard (29 CFR 1910.134, Respiratory Protection, requires Mount Sinai to develop a written Respiratory Protection Program, which describes required worksite-specific procedures and elements for required respirator use. The Respiratory Protection Program mandated by OSHA requires Mount Sinai to provide respirators that protect employees from inhalation of hazardous materials that cannot be controlled by engineering or administrative controls. Mount Sinai must provide effective annual training to employees who are required to use respirators. Prior to working in areas requiring respiratory protection, employees must:

1. Complete the Respirator Medical Evaluation Questionnaire (for first-time fit-test applicants) or the Periodic Medical Questionnaire (for reevaluation of existing respirator users), which is then reviewed by Workforce Health;
2. Undergo medical evaluation by Workforce Health; and
3. Complete fit testing for tight-fitting N95 respirators on an annual basis.

Employees should visit the Environmental Health and Safety website to obtain information on the institutional Respiratory Protection Program, including Medical Evaluation Questionnaire and details on N95 respirator fit testing sessions.