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Protection of Vacuum Systems Laboratory work involving culturing and preparation of cells and tissues generates liquid biohazardous waste that requires careful management of disinfection and disposal. Proper disinfection techniques and safe disposal of biohazardous materials is critical for safety both inside and outside the facility. Guidelines established by the Centers for Disease Control and Prevention (CDC) and the National Institute of Health (NIH) Guidelines for Biosafety Level 2 (BSL-2) state that vacuum lines should be protected with an in-line HEPA filter, or an equivalent, along with disinfectant traps to prevent the release of biohazardous aerosols into the laboratory or building vacuum system. This information is pertinent for all labs that work with cell and tissue culture. Adhering to the proper setup and usage of laboratory vacuum lines will help protect the building’s vacuum system and any personnel that service the equipment from biohazardous materials.

All principal investigators, laboratory supervisors and laboratory personnel are responsible for the proper decontamination and disposal of biohazardous liquids and materials in accordance with the information found on this fact sheet as well as the guidelines included in the EHS hazardous waste program. For more information, visit http://www.ehs.neu.edu/hazardous_waste/fact_sheets/hazardous_waste

Lab personnel must wear the appropriate PPE when handling biohazardous wastes and while working in the lab (e.g. proper gloves, eye protection, lab coats, etc). !

(A)!Collection!flask!–!collects! contaminated!fluids!and!allows! contact!with!disinfectant!(e.g.!10%! bleach)!(B)!Overflow!flask!–!collects! overflow!from!the!collection!flask! and!!minimizes!biological!aerosols! (C)!InCline!HEPA!Filter!–!prevents! contamination!of!vacuum!system! (D)!vacuum!port! !

The vacuum setup above consists of the following components:!

• 2 vacuum flasks (plastic preferred*) • Thick walled plastic tubing • Rubber stoppers • In-line HEPA filter, or equivalent**

* The use of plastic flasks is recommended to prevent breakage/spill. Fisher offers Nalgene plastic products as an alternative to glass flasks (more information below) ** Filter must be in-line, hydrophobic, made of PTFE and have a particle retention of 0.3 µm (more information below) ! Aspirator bottles or suction flasks should be connected to a vacuum flask (A) for the collection of contaminated liquids. Vacuum flask (A) is attached to a second vacuum flask (B). The second flask ensures that the capacity of the first flask does not exceed its limit and the tube should be submerged in liquid to minimize aerosol formation. Each flask must contain an appropriate disinfectant, such as bleach. An in-line HEPA filter, or equivalent, is attached after vacuum flask (B) before the vacuum port. The filter must be replaced when needed (visible blockage, notable resistance, etc). Replaced filters are considered hazardous waste and shall be disposed of as biohazardous waste.

The vacuum setup should be in a biosafety cabinet. If not, the setup must be placed at floor level in a high-walled, secondary container that can accommodate both flasks. Placement in a secondary container can prevent accidental breakage and the release of biohazardous contents.

Ensure that all flasks are labeled. Labels on the flasks must include the type of chemical disinfectant used, the concentration and any other contents. If the flask contains no disinfectant, the flask must be labeled as ‘biohazardous’. Do NOT use EHS hazardous waste labels to label flasks.

Disinfecting

For inactivating purposes, ensure that the biohazardous material is in contact with 10% bleach for 30 minutes, or an approved disinfectant for the appropriate contact time. As long as there are no chemical or radiological hazards present in the solution, the liquid can be disposed of in the sink after the contact time has been reached. Ensure that sufficient water is used to flush the liquid material through the pipes. If chemical/radiological hazards still exist in the waste solution, please refer to the hazardous waste management program for proper disposal (see http://www.ehs.neu.edu/hazardous_waste/ for more information).

Ordering Information

Materials mentioned in the setup are available for purchase at Fisher Scientific https://www.fishersci.com/us/en/home.html

Filters ! Name Description (from Dimensions Part # Fisher!Catalog! Fisher) No! Whatman mildly hydrophobic, Inlet/outlet: 1/4 to Whatman 09-744-79 HEPA-Vent 0.3µm particle 3/8 inch stepped 6723-5000 Filter retention, barb autoclavable

! Plastic Flasks Thermo Scientific™ Nalgene™ Polypropylene Vacuum Flask

Capacity! Stopper!Number! Accepted!Tubing! Autoclavable! Fisher!Catalog!No!

500!mL! 7! 1/4!B!3/8! Yes! 10B182B50A!

1000!mL! 8! 1/4!B!3/8! Yes! 10B182B50B!

1700!mL! 10! 1/4!B!3/8! Yes! 10B182B51!

Plastic Tubing Thermo Scientific™ Nalgene™ 180 Clear Plastic PVC Vacuum Tubing

Diameter!(inner)! Diameter!(outer)! Autoclavable! Fisher!Catalog!No!

1/4! 5/8! Yes! 14B176B24!

3/8! 7/8! Yes! 14B176B42!

Rubber Stoppers Fisherbrand™ One-Hole Rubber Stoppers

Stopper!#! Flask!Compatibility! Fisher!Catalog!!No! 7! 10B182B50A!(500!mL)! 14B135L! 8! 10B182B50B!(1000!mL)! 14B135M! 10! 10B182B51!(1700!mL)! 14B135P!