Standard Operating Procedure (SOP)

1.      Process

Ethidium bromide is commonly used in molecular biology laboratories as a nucleic acid material which fluoresces a red-orange color under UV light with increased fluorescence to double stranded DNA.

2.      Physical Hazards

q       In powder form, fire is possible at elevated temperatures.

q       Incompatibilities and reactivities: strong oxidizers.

3.      Health Hazards

q       Moderately toxic after acute exposure.

q       Irritating to the skin, eyes mouth and upper respiratory tract.

q       Can cause heritable genetic damage.

4.      Personal Protective Equipment

q       EYE PROTECTION

·          Safety glasses, goggles or face shields shall be worn during operations in which ethidium bromide might contact the eyes (e.g., through vapors or splashes of solution). 

·          Ordinary (street) prescription glasses do not provide adequate protection.  Adequate safety glasses must meet the requirements of the Practice for Occupational Education Eye and Face Protection (ANSI Z87.1-1989) and must be equipped with side shields. 

·          Fisherbrand® Chemical Splash Goggles, catalog # 19-041-739 are recommended.

·          Contact lenses shall not be worn if the potential exists for ethidium bromide exposure.

q       HAND PROTECTION

·          Use of Nitrile gloves are recommended for use against permeation by ethidium bromide and may provide protection for more than 4 hours but fewer than 8 hours.  Lab Safety Supply catalogue # 2BD-1414655.

q       LAB COATS, ETC.

·          Lab coats, close toed shoes and long sleeved clothing shall be worn when handling ethidium bromide. Protective clothing shall be worn to prevent any possibility of skin contact with ethidium bromide.

q       SAFETY SHOWER/EYEWASH

·          Where the eyes or body of any person may be exposed to ethidium bromide, suitable facilities for quick drench or flushing of the eyes and body shall be provided within the work area for immediate emergency use. 

·          Bottle type eyewash stations are not acceptable.

5.      Designated Area for Use and Containment Devices

q       All ethidium bromide work shall be done in the laboratory fume hood.

q       The fume hood’s sash must be in the position where a face velocity of 100 feet per minute is achieved.  

6.      Special Handling Procedures and Storage Requirements

q       Keep away from ignition sources.

q       Keep away from strong oxidizers.

q       Spill-X-S Kit, which is available through Lab Safety Supply, catalog #2BD-14628, must be available in the event of a liquid spill less than 300 cc.

7.      Waste Disposal

q       Excess ethidium bromide and all waste material containing ethidium bromide must be placed in an unbreakable secondary container labeled with the following "HAZARDOUS WASTE ETHIDIUM BROMIDE ". Full containers of ethidium bromide waste must be disposed of according to WFUHS EH&S Hazardous Waste Program.

q       Ethidium bromide gels must also be disposed of according to WFUHS EH&S Hazardous Waste Program.

q       To reduce ethidium bromide waste in solution, recommend use of the Greenbag kit manufactured by BIO 101. Their website address is http://www.bio101.com/. The green bag must be disposed of through the WFUHS EH&S Hazardous Waste Program.

8.      Decontamination of Equipment and Area

      Solutions shall be absorbed and area decontaminated with soap and water.

9.      Spill and Accident Procedures

q       Remove all sources of ignition if it involves the ethidium bromide powder form. Then use water to lessen dust formation and absorb.

q       Hold all solid absorbent material for disposal through WFUHS EH&S Hazardous Waste Program.

q       In the event of a liquid spill greater than 300 cc, immediately vacate and secure the area. Contact WFUBMC Security at 716-9111.