These safety training resources, prepared solely for the use of the Regents of the University of California, were provided by a variety of sources. It is your responsibility to customize the information to match your specific operations. Neither the University of California nor any of its employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise, does not necessarily constitute or imply its endorsement, recommendation, or favoring by the University of California. The views and opinions of authors expressed herein do not necessarily state or reflect those of the University of California, and shall not be used for advertising or product endorsement purposes.
Chemical Use in Animal Research ● Standard Operating Procedures
(ENU) CAS# 759-73-9
X LAB
Date SOP Created: Draft 7/15/10
Instructions: Insert specific details pertaining to your research; contact EH&S or OLAC as needed for assistance.
Hazard information ENU is toxic, carcinogenic, mutagenic, and teratogenic. ENU may cause cancer and heritable genetic damage. ENU may cause harm to the unborn child. ENU is harmful if swallowed, inhaled, or absorbed through the skin. Hazardous waste No. U176; NIOSH
No. YT 3150000; SAFDS* 0001; IMEMDT 0016
Preparation Purchased from Sigma-Aldrich (N-3385-1G) in 1 gram isopacs. Order ENU isopacs only as needed and use as soon as possible. Store sealed and labeled hazardous, toxic, potent mutagen”. Never handle ENU powder. ENU should be dissolved within the isopac bottle by injecting solvent into the isopac. All procedures must be performed in a chemical fume hood in Room: X Laboratory personnel must wear a laboratory coat, safety goggles, and gloves. Wear gloves at all times and change them whenever you suspect contact with mutagen or whenever you leave the fumehood.
Transportation
Use
ENU is shipped by Sigma-Aldrich in a sealed container as a 1-gram isopac. On the day of the mutagenesis, the container is carried to the chemical fume hood and opened so that the sealed isopac can be retrieved. Whenever transporting between rooms or buildings, the outer package should clearly state the chemical name and the PI name.
Injections will be performed in a chemical fume hood in Room: X over absorptive paper .
Signage must be posted as described below. ENU is solubilized within the isopac bottle by injecting solvent into the isopac with a syringe through the rubber seal. The required amount of solubilized ENU is withdrawn using a plastic syringe and injected into the appropriate mice. The final concentration of ENU used in the procedure is X . Cages containing animals injected with ENU must be clearly labeled and indicate that animal technicians should not change them.
Disposal
If unsure, contact EH&S at 642-3073 to determine if chemical disposal is necessary for bedding, unused drinking water, and/or carcasses following treatment.
ENU is unstable and should disappear rapidly in the aquatic environment. Because ENU is an alkylating agent, it can be inactivated by treatment with sodium thiosulfate and high pH (10% sodium thiosulfate, 1% sodium hydroxide (~pH 10). The half-life of ENU in
10% sodium thiosulfate in basic solution is about 10 minutes at 37°C. We inactivate almost all objects (gloves, pipets, containers) in 10% sodium thiosulfate, 1% sodium hydroxide for 24 hours in the fume hood where the chemical mutagenesis procedure is performed.
In the hood, inactivate the mutagen by pouring the treatment and rinse solutions into an equal volume of 20% sodium thiosulfate, 2% NaOH and let sit for 24 hours. Hardware that comes in contact with mutagen (mouse cages, pipets, pipet tips, etc.) should be placed in a separate, pre-prepared 10% sodium thiosulfate, 1% NaOH bath for 24 hours.
After 24 hours, the solution can be poured down the sink. Some materials (absorptive paper, for example) must be bagged and collected for disposal by EH&S. Dispose of animals that die during the first 24 hours by placing them into the sodium thiosulfate
Office of Environment, Health & Safety ● http://ehs.berkeley.edu/ ● 642-3073
OLAC Manager Initials/Date: _______________ Effective Dates: ____________ to ____________

These safety training resources, prepared solely for the use of the Regents of the University of California, were provided by a variety of sources. It is your responsibility to customize the information to match your specific operations. Neither the University of California nor any of its employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise, does not necessarily constitute or imply its endorsement, recommendation, or favoring by the University of California. The views and opinions of authors expressed herein do not necessarily state or reflect those of the University of California, and shall not be used for advertising or product endorsement purposes. inactivation bath, then bag and dispose in the biohazardous waste barrel in cold room.
After 24 hours, carcasses can be disposed of in a normal manner. 24 hours after injection with ENU, the lab must transfer animals to a fresh cage within a biosafety cabinet in
Room X. The bedding in the old cage will be transferred to a plastic bag containing waste paper saturated with inactivating solution. After 24 hours the bag should be disposed in the biohazardous waste barrel in the cold room. The cage itself should also be treated with inactivating solution prior to being returned to OLAC.
. To arrange for waste pickup by EH&S, complete an electronic Material Packing List (eMPL) form available at http://mpl.ehs.berkeley.edu/ and submit it to EH&S. Make sure you follow the instructions when packaging the waste for pickup.
Spill response and emergency procedures
ACCIDENTAL RELEASE MEASURES: See MSDS
In case of a spill of ENU powder or ENU solution, notify other laboratory personnel immediately and ask them to leave the laboratory. The door to the room containing the fume hood should be closed. If a clean-up is necessary outside of a fume hood, call
EH&S. The powder will be carefully swept up, placed in a bag, and held in the chemical hood for waste disposal or inactivation. Avoid raising dust. Ventilate area and wash spill site after material pickup is complete. If ENU solution is spilled, it should be contained within the chemical hood. If any spills outside the chemical hood, it should be absorbed using absorbent paper. Paper should be bagged in a hazardous chemical waste bag and stored in the hood for EH&S pickup. Ventilate area and wash spill site.
For emergency clean up response, notify EH&S at 642-3073.
Hazard communication, signs, cage cards, etc.
Several days before the mutagenesis, notify facility personnel and members of the laboratory of the time and place of mutagenesis. On the day of the procedure, post signs on the fume hood and on the door to the room: "Restricted Entry -- Chemical
Mutagenesis in Progress." Have the OLAC manager review and initial below prior to posting this SOP when ENU is in use.
All researchers handling this material should read and sign this document. When animals have been treated with/administered this chemical, all cage cards should be labeled appropriately and this SOP should be posted in a plastic sleeve on the applicable cage rack or housing door to notify OLAC staff and other personnel.
Unique instructions FIRST AID MEASURES: If swallowed, wash out mouth with water if person is conscious and call a physician. If inhaled, remove to fresh air. If breathing becomes difficult, call a physician. In case of skin contact, flush with copious amounts of water for at least 15 minutes. Assure adequate flushing by separating the eyelids with fingers.
Wash contaminated skin with soap and water, taking care to aviod rubbing or raising the skin temperature. Call a physician. FIRE FIGHTING MEASURES: See MSDS,
For urgent care call 911 or go to the Tang Center or after hours Alta Bates hospital.
Additional information or references
*LABORATORY PERSONNEL MUST READ THE FOLLOWING
INFORMATION*(1) mutagenesis protocol,
MSDS (CAS # 759-73-9),
(3) .
(2) Scientific reference for
Refer to the MSDS available at
Office of Environment, Health & Safety ● http://ehs.berkeley.edu/ ● 642-3073
OLAC Manager Initials/Date: _______________ Effective Dates: ____________ to ____________

These safety training resources, prepared solely for the use of the Regents of the University of California, were provided by a variety of sources. It is your responsibility to customize the information to match your specific operations. Neither the University of California nor any of its employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise, does not necessarily constitute or imply its endorsement, recommendation, or favoring by the University of California. The views and opinions of authors expressed herein do not necessarily state or reflect those of the University of California, and shall not be used for advertising or product endorsement purposes. http://www.ehs.berkeley.edu/healthsafety/msds.html, your lab’s chemical hygiene plan, or contact your supervisor or EH&S at 642-3073 for further guidance.
Training documentation (name and date):
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Office of Environment, Health & Safety ● http://ehs.berkeley.edu/ ● 642-3073
OLAC Manager Initials/Date: _______________ Effective Dates: ____________ to ____________