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Department of Biochemistry
School of Molecular Sciences Safe Work Procedure
ETHIDIUM BROMIDE
Use
Ethidium bromide is added to electrophoresis gels for visualisation of
nucleic acids.
Hazards
Class 6 – Toxic. Potent mutagen.
Risk Control Measures
Only use Ethidium bromide (EtBr) after receiving safety training
(Laboratory Induction & Authorisation). Wear safety glasses when using
ethidium bromide. Avoid skin contact, ethidium bromide may be absorbed
through the skin. Wear latex gloves and laboratory coat. Always dispose
of gloves after use. Don't touch equipment, door handles, phone,
keyboard, etc.
Weighing solid - Powder may cause irritation when inhaled - wear dust
mask and use in ventilated area. Use designated micropipette only when
dispensing the liquid.
Engineering / Ventilation Controls
Ensure access to an eyewash facility in areas where ethidium bromide
is used. Preferably weigh the solid in a fume hood.
Storage Requirements
Store in a cool, dry place away from strong oxidizing agents. Keep
containers tightly closed. Use with adequate ventilation.
First Aid / Spill Control Procedures
Wash off immediately with copious amounts of cold water (at least 10
minutes). Ethidium bromide is absorbed through the skin so follow the
cold water washing with a thorough washing with warm water and soap.
Contaminated clothing should be removed as soon as possible and
thoroughly washed.
In case of contact with eyes, immediately flush eyes with copious
amounts of water for at least 15 minutes (eyewash).
Seek medical attention.
Poisons Information Centre (0 13 1126).
If the spill is on equipment, use ultraviolet light (wear appropriate
eye protection) to locate spill, then use the decontamination procedure
outlined below.
Wear protective clothing.
Small spill:
If in solution, absorb freestanding liquid using vermiculite or Polyzorb
from Spill Kit. Use ultraviolet light to locate spill. Follow
instructions on Spill Kit.
Large Spill: Notify others in the area of spill. Evacuate area. Call
the Molecular Sciences Office (9479 2160) or Zisis Pouros (9479 1294) for clean up.
Barricade area with tape to prevent entry until arrival of response
personnel. Provide assistance and information to spill clean up crew.
Waste
Store waste: Liquid - In proper waste container
Solid - In proper waste container
Contact the Molecular Sciences Office (9479 2160) for current waste disposal
instructions.
Ethidium Bromide Liquid Disposal
1. Add 10g activated charcoal per 2.5L waste.
2. Leave for 1 hour, with occasional shaking.
3. Filter contents through Whatman Number 1 filter paper.
4. Filtrate may be disposed of down the sink.
5. Charcoal & paper is treated as solid hazardous waste and disposed in
the EtBr Solid Waste Bucket.
(Adapted from Bensaude (1988) Trends In Genetics 4(4), 89 – 90)
Staining gels
During electrophoresis – After boiling up the agarose, let it cool
down before adding EtBr
Afterwards -Soak gel in a well marked plastic container - Put name and
date on container as it is possible to re-use the staining solution.
Applicable Standards and Regulations
Occupational Health and Safety (Hazardous Substances) Regulations
1999
Australian Standard AS 2243.10 (Safety in laboratories - Storage of
chemicals)
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