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What Is the Scope of This Guideline?
This guideline applies to all lab personnel working with ethidium bromide (EB) (CAS# 1239-45-8). Ethidium bromide is a powerful mutagen and is a skin, eye and respiratory irritant. It can be readily absorbed through the skin. Avoid direct skin contact!
How Do I Protect Myself?
Review the material safety data sheet (MSDS) prior to using this material.
Personnel handling ethidium bromide must wear adequate eye protection. Adequate safety glasses must meet the requirements of the Practice for Occupational and Educational Eye and Face Protection (ANSI Z.87. 1 1989) and must be equipped with side shields. Ordinary prescription glasses do not provide adequate protection unless they meet this standard and are marked as such. Safety glasses with side shields do not provide adequate protection from splashes; therefore, when the potential for splash hazard exists other eye protection and/or face protection must be worn (i.e., splash goggles).
Ensure that UV protective eyewear is utilized when visualizing ethidium bromide.
Gloves should be worn when handling ethidium bromide. Disposable nitrile gloves provide adequate protection against accidental hand contact with small quantities of most laboratory chemicals. Double-glove if extended work or exposure is expected.
Appropriate protective clothing should be worn if the possibility of skin contact is likely. Open-toe shoes are not appropriate when handling chemicals in a laboratory or in other areas where there is a potential for exposure.
Always wash your hands after handling this material.
Engineering, Ventilation and Administrative Controls
Brandeis employees who work with hazardous chemicals must be apprised of the hazards of chemicals present in their work area. This training must be provided before initial assignment to a lab and distribution of keys or must work under supervision before training occurs. Lab personnel need to be trained to properly handle ethidium bromide, the location of this material in the lab, the proper measure to handle and store it and measures to be used to clean up spills.
Any procedure which could result in the production of ethidium-bromide aerosols should be done in a fume hood.
Proper safety precautions must also be taken when dealing with UV light.
Special Handling and/or Storage Requirements
Store ethidium bromide away from strong oxidizing agents in a cool, dry place, and ensure that the container is kept undamaged and tightly closed.
Ehidium bromide should be used in a specially designated area. All lab personnel should be made aware through training or appropriate hazard warnings (signs/labels) that EB is used in this area.
Handling Emergencies Involving Ethidium Bromide
Anticipate emergency situations, have proper handling equipment in the lab and readily available for spills. Check the MSDS to determine what is appropriate.
In the event of a spill or adverse reaction, notify lab personnel immediately that an incident has occurred. Do not attempt to handle a large spill/reaction/fire, or one in which you are not trained or equipped for. Turn off all ignition sources if this can be done safely; vacate the area and call for assistance.
Laboratory emergencies should be reported to the public safety office at ext. 6-3333. Public safety will also contact the Office of Environmental Health and Safety at ext. 6-4262. Communicate the following:
- Location of spill/incident
- Type of material involved and quantity
- Injuries involved
- Your location/contact information (or who to contact for further information)
Notify the principal investigator or designated safety officer as soon as possible.
If exposed to EB, skin/eye contact, immediately flush the eyes or skin with copious amounts of water for at least 15 minutes or until medical help arrives and you are instructed otherwise.
If EB dust is inhaled, move the victim to fresh air and contact public safety immediately for medical assistance.
Notify the Department of Environmental Health and Safety of significant ethidium-bromide spills.
Always wear full protective clothing, as described above, during any cleanup procedure. You should only clean up spills if you are aware of the hazards and have the proper safety and cleanup equipment.
A spill of EB powder can be carefully wiped up with wet paper towels the area decontaminated. Utilize UV light to locate the spill.
A spill of EB in liquid form should be absorbed with dry paper towels. Use a UV light source and illuminate the area to locate any remaining EB. All cleanup material should be placed in containers for proper disposal. DO NOT place EB-contaminated material in the regular trash.
Waste Disposal Requirements
All materials contaminated with ethidium bromide should be packaged similar to hazardous waste at Brandeis and disposed of through the Brandeis Hazardous Waste Program as a special waste. If you do not have a waste container, contact the hazardous waste hotline at ext. 6-2561 and leave a message, including your name, location and question.
Disposal requests should be called into the Waste Hotline at ext. 6-2561.
Wash hands and arms with soap and water immediately after handling any chemical.
Clean work areas carefully when done. Dispose of contaminated material in a hazardous waste storage container. Prepare and utilize the following solution to decontaminate lab surfaces.
Prepare the decontamination solution just prior to use. The solution is acidic, so wear full protective equipment (lab coat, gloves and goggles, not safety glasses) when preparing or using the decontamination solution. The solution consists of:
- 4.2 g of sodium nitrite (NaNO2, CAS # 7362-00-0)
- 20 ml of hypophosphorous acid (50%) (H3PO2, CAS # 6303-21-5)
- 300 ml of water
- Wash the area with a paper towel soaked in decontamination solution. Then rinse the area five times with paper towels soaked with tap water, using a fresh towel each time.
- Soak all the towels in decontamination solution for one hour. Then remove them, gently wring out excess solution, place in a sealed bag or container.
- Using a UV light, check the area to ensure that all the EB has been removed.
- Contact the Brandeis waste hotline for disposal of the decontamination solution, towels and gloves used during the cleanup.
- If the acid may damage the contaminated surface, use a few additional rinses. Again, soak all the towels in decontamination solution for at least one hour before disposal.