Purpose of this SOP:

To ensure that every member of the iGEM Concordia 2016 team has an understanding of the project and the safety information associated with it. This SOP contains information on the safe handling of nanoparticles, as well as the various chemicals being used in this project. Protocols for each part of the project involving nanoparticle synthesis and attachment to cells are also included in this document.

Responsibility:

It is the responsibility of each member of the iGEM Concordia 2016 team to read this document and confirm that they have understood it. Each member must provide their signature on the last page of this document upon reading and understanding this SOP.

The following is general information for safely handling of nanoparticles (Click on the boxes below for the specific SOP of each section of this project):

Personal Protective Equipment (PPE)

-Wear nitrile gloves, do not allow nanoparticles to come in direct contact with skin.

-Remove gloves when leaving the lab. DO NOT touch doorknobs with your gloves on.

-If gloves came into contact with nanoparticles, dispose in designated solid nanoparticle waste container.

-Otherwise, dispose of gloves in biohazardous waste bin.

-Do not touch personal items such as phones with gloves, as this may transfer nanoparticles or reagents to the item.

-Wear safety glasses to prevent eye exposure to harmful materials.

-Wear a lab coat while working in the lab; do not take the lab coat outside of the lab or bring it home to be washed. There are designated areas for cleaning lab coats.

-Wear closed-toe shoes (i.e. no sandals).

-People with long hair are to tie their hair back when working in the lab.

Preventative Measures

-Allocate a specific lab area for working with nanoparticles to limit exposure to the rest of the lab.

-Work in:

-A fume hood for nanoparticle synthesis and liquid nanoparticle waste disposal. The fume hood that our team is using is located in the Chemical Preparation room GE300.04.

-A biosafety cabinet for nanoparticle attachment to cells. The biosafety cabinet being used by our team is located in the Cell Culture room in GE330.00.

-Attend appropriate biosafety training sessions: WHMIS 1988, WHMIS 2015, Disposal of Hazardous Waste, Biosafety Training,  and Safe Handling of Nanomaterials.

-Work with nanoparticles in suspension instead of powder form to decrease risk of inhalation.

Keep work area clean and avoid spills. In case of a spill, contact security (514-848-3717 - dial 1 for urgent situations; dial 2 for non-urgent situations).

Keep separate containers for disposal of waste containing nanoparticles; these containers should be properly labeled.

When moving nanoparticles around the lab, keep the material in sealed containers (use aluminum foil if there is no appropriate cover for the container being used). If transporting the nanoparticles to a different room, place container inside a bin in case of spill.

All solutions containing nanoparticles should be properly labelled. Label should include: the type of nanoparticle (ie. gold), date of synthesis, synthesis method, concentration (if possible), size (range) of nanoparticles, initials of the team members who performed the synthesis or name of company it was purchased from.

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Precaution for safe handling of hazardous materials

When handling materials that are potentially hazardous, one needs to be well aware of the special handling procedures indicated in the Safety Data Sheet (SDS)  for that specific item. To do this, one should become familiarized with any pertinent SDS upon receiving a chemical, then once again look through the SDS before its usage. Also knowing where all safety equipment is stored and having it readily accessible if required, is crucial. The correct updated telephone numbers of emergency contacts should be posted in a visible location in the lab in the case of a chemical spill or accident.

While working, handling, and cleaning hazardous chemicals, ensure that you:

-Avoid physical contact with the material.

-Do not leave the material exposed and unattended.

-Avoid production of aerosols and dusts with the chemical to prevent inhalation of the material.

-Ensure that the proper exhaust ventilation systems are functional.

-Take precautions to avoid fire hazards with the material (ie. do not smoke in the laboratory, do not leave -light or ignition sources exposed within the vicinity of the chemical).

-Do not eat or drink in the laboratory.

-Do not physically damage chemical containers or use those that are damaged.

-Use of nitric acid should not be performed unless accompanied by another trained laboratory member; nitric acid is highly dangerous, therefore great attention should be made to preventing nitric acid spills.

Storage procedures

-When storing hazardous materials, ensure that the respective storage location is not obstructed, is placed away from the working bench and other commonly used materials, is clearly identified with warning signs, and is not placed near chemicals with which it may react.

-Before storing hazardous materials, verify that the container enclosing the materials is intact, not damaged, and properly labelled following WHMIS workplace label requirements.

-When storing toxic chemicals, make sure that the materials are not left open, unlabeled, or unattended when in use.

-Dispose of all expired chemicals by following the correct waste disposal procedures.

-Regularly verify that storage areas are up to par by making sure that containers and cabinets are clean and that there are no container leakages, damages, or deficiencies in storage equipment - if the storage area is not up to par, then fix any and all deficiencies as quickly as possible.

-Containers containing chemicals and lab materials should be stored at a reasonable height for handling so that each lab member can easily access the materials (reasonable height would be below eye level) - storing materials higher up increases the risk of spills, container damage, dropping of materials, physical injury and chemical exposure.

-Nanoparticles should be stored in the 4°C fridge or at room temperature. Should be in a sealed container and labelled with the date, method of synthesis, type of nanoparticle and names of the individuals who synthesized the nanoparticles.

Waste disposal

-Liquid waste is to be poured into ‘Liquid chemical waste containers’ (depending on the type of liquid) - do NOT fully fill the liquid chemical waste containers as this may cause spills (leave 10-20% of free space to allow solvent expansion). When a container is ~80% full, contact Environmental Health and -Safety (EHS) and provide information on the type of waste and the location of the waste. EHS will ensure proper disposal of the waste.

-Do not pour toxic chemicals and products down the sink/drain.

-Place any non-biohazardous solid waste that has come into contact with nanoparticles, such as used gloves and contaminated paper, in the ‘ nanoparticle solid waste chemical containers’. Make sure the type of waste is properly identified (ex. Nanoparticle contaminated solids).

-In the case of non-contaminated broken glass, such as beakers and flasks, immediately sweep up and dispose of the glass using a broom and dustpan, then place into designated ‘broken glass’ cardboard containers - do NOT touch the broken glass with hands even if wearing gloves.

-All sharps (such as syringes, needles, scalpels, pipettes, broken glass) which are contaminated with chemicals must be discarded in puncture-proof solid chemical waste containers - do NOT bend, shear, recap, or remove needles from syringes and do NOT place sharps in regular garbage.

-Nonhazardous, non-nanoparticle waste materials may be discarded into regular garbage containers.

-Discard materials that have been used and any waste produced, to prevent accumulation and clutter on working benches. Ensure that materials are disposed of properly.

-Expired chemicals and empty chemical bottles can be given to EHS for disposal, given that the bottles are in good condition and can be properly identified. Ensure that the expired chemicals remain in their original container. Do NOT combine expired chemicals with others in different waste containers.

-Empty solvent bottles that have been properly cleaned and rinsed with water can be placed next to regular garbage bins. These will be picked up by janitors. Bottles and glass containers used to contain chemicals cannot be recycled.

-Liquid nanoparticle waste must be disposed of in the chemical waste container labelled ‘liquid nanoparticle waste’. This container is found in the fume hood. Once this container is 80% full, contact EHS to pick up the waste container.

-Nanoparticle coated cells should be disposed of in biohazardous waste containers through a series of steps as follows: centrifuge the cell/nanoparticle solution and dispose of the supernatant in the ‘liquid nanoparticle waste’ container - the pellet should then be disposed of in the biohazardous waste containers.

Hazardous chemical spill

-In the case of a minor chemical spill, proper cleanup procedures must be followed using SDS instructions. If unsure or the spill cannot be managed using available tools, do not attempt to manage the spill on your own and seek help (see next point).

-Major accidents, incidents, and injuries are to be reported to security (514-848-3717 - dial 1 for urgent situations; dial 2 for non-urgent situations), then to EHS (514-848-2424 ext. 4877), followed by the lab supervisors and advisors, then finally the Biology department administrator (ext. 3400).

-During working hours, any minor incidents or accidents in the lab are to be reported to the safety officer (ext. 4877), the lab supervisors or the Biology department administrator (ext. 3400).

Cleanup procedure upon completion of experiment

-Remove any respirators, face shields, or masks.

-Discard all waste and used chemicals in their appropriate bins.

-Any and all lab instruments used are to be properly placed in the sink for washing or placed in their appropriate bins for cleaning - other equipment that does not require cleaning is to be placed back to its respective storage location.

-Remove and properly arrange materials at the bench. Once the bench is clear, clean it with 70% ethanol.

-If working in a fume hood or biosafety cabinet, remove and properly discard of all waste and clear the area of lab instruments by placing them in their respective storage locations; clean the base of the fume hood/biosafety cabinet; close the light in the fume hood and the biosafety cabinet; close the fume hood/biosafety cabinet sash.

-Remove lab safety goggles.

-Take off gloves using the proper glove removal technique by folding gloves inside out to avoid skin contact with outside surface of the used glove.

-Remove labcoat and store properly.

-Wash hands with soap then dry, once all PPE has been removed (hand washing should be done whenever exiting the lab).

-Ensure that no machinery has been left ON that should not be, when the lab is not being supervised.

-Exit the laboratory.

-Work clothes should be laundered separately from other clothing.

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