Standard Operating Procedures Laboratory Specific Chemical: Bromo-Deoxyuridine (BrdU) Please fill out the form completely. Print a copy and insert into your Laboratory Safety Manual and Chemical Hygiene Plan. Refer to instructions for assistance. _____________________________________________________________________________ Department:________________________ Date when SOP was written:_______ Date when SOP was approved by the lab supervisor: ___________________ Principal Investigator:___________________________________________________ Internal Laboratory Safety Coordinator/Lab Manager:___________________________________ Laboratory Phone:____________________ Office Phone:_____________________ Emergency Contact:____________________________________________________ (Name and Phone Number) Location(s) covered by this SOP:__________________________________________ (Building/Room Number) _____________________________________________________________________________ Type of SOP: Process Hazardous Chemical Hazardous Class Purpose Bromo-Deoxyuridine (BrdU) is used to label cells undergoing S-phase synthesis. It is a thymidine analogue and gets incorporated in DNA. This SOP describes the safe usage and handling, and disposal of BrdU. Physical & Chemical Properties/Definition of Chemical Group Chemical Name: 5-Bromo-2’-Deoxyuridine (BrdU) CAS#: 59-14-3 Class: Reproductive toxin Molecular formula: Molecular weight: Form: C9H11BrN2O5 307.13 g solid; white needles when crystallized from absolute ethanol; no distinctive odor 1 Solubility: Volatility: Some solubility in water or saline Stable dark and heat/humidity for 3 mos. at temperatures below 60°C. Sunlight causes discoloration. Frozen aqueous solutions exposed to UV result in formation of debrominated dimmer. Photooxidation occurs in air/light. BrdU is hydrolyzed at the N-glycosyl bond, this rate increases at alkaline pH. Flammability restrictions do not apply. Other: Laboratory operations should be conducted in a fume hood BrdU is a probable combustible compound. BrdU is cytotoxic, teratogenic, and mutagenic. It is not carcinogenic; it is a topical antiviral agent and is useful in the treatment of neoplasms and sensitizes tissue to X-rays. Potential Hazards/Toxicity Toxic Effects: BrdU is a toxic substance. Acute Effects: Wheeze and cough, shortness of breath, burning in the mouth, throat, or chest. Chronic Effects: Reproductive disorders and genetic alterations. BrdU is incorporated into tissue DNA in place of thymidine in animals. BrdU-substituted DNA replaces normal DNA and chromosomal proteins are altered through: chromosome lengthening, chromatid breakage, and changed sister chromatid exchange frequencies. Meiosis and mitosis are affected. Carcinogenic Effects: Brdu is suspected to be a carcinogen resulting in heritable genetic damage. It is harmful to the unborn child. Mutagenic/Teratogenic Effects: BrdU is not mutagenic—Ames test; but BrdU is mutagenic— micronucleus and sperm abnormality assay. BrdU is a strong teratogen in rodents. Systemic effects: BrdU integrates into DNA and affects the colon, stomach, bone marrow, and spleen. Personal Protective Equipment (PPE) Wear double nitrile gloves, full-length lab coat, and safety glasses/face shield in addition to long pants and closed-toe shoes. Always wash hands after removing gloves following handling BrdU. Engineering Controls Always handle BrdU inside a certified chemical fume hood or a ducted biosafety cabinet. First Aid Procedures Skin & Eye Exposure: Skin: remove contaminated clothing and wash skin with soap and water; avoid rubbing and increases in temperature. Seek medical attention promptly. Eye: 2 rinse immediately with copious amounts of running water for at least 15 minutes. Seek attention of a licensed Ophthalmologist promptly. Ingestion: Drink lots of water/milk. Induce vomiting. Seek medical attention (refer gastric lavage). Inhalation: Remove rapidly to clean air. Administer rescue breathing if necessary and call emergency services. Seek medical attention if needed. Special Handling and Storage Requirements Whenever feasible, procedures with the potential for producing BrDu aerosols should be conducted with a certified chemical fume hood. Needles used for BrDu injection will be disposed of in approved sharps containers immediately following use. Needles used for BrDu injection should never be bent, sheared, or recapped. Spill and Accident Procedure In case of eye exposure: go to eyewash station and rinse eyes for 15 minutes, then seek medical attention If inhaled: remove to fresh air, seek medical attention immediately If absorbed: through skin, immediately flush with plenty of water, remove contaminated shoes and clothing, seek medical attention immediately If ingested: Do NOT induce vomiting, loosen tight clothing, and seek medical attention immediately. Chemical Spill Dial 911 and x59797 Spill – Help contaminated or injured persons. Evacuate the spill area. Avoid breathing vapors. Eliminate sources of ignition if the chemical is flammable. If possible, confine the spill to a small area using a spill kit or absorbent material. Keep others from entering contaminated area (e.g., use caution tape, barriers, etc.). Small (<1 L) – If you have training, you may assist in the clean-up effort. Use appropriate personal protective equipment and clean-up material for chemical spilled. Double bag spill waste in clear plastic bags, label and take to the next chemical waste pick-up. Large (>1 L) – Dial 911 (or 310-825-1491 from cell phone) and EH&S at x59797 for assistance. Chemical Spill on Body or Clothes – Remove clothing and rinse body thoroughly in emergency shower for at least 15 minutes. Seek medical attention. Notify supervisor and EH&S at x59797 immediately. 3 Chemical Splash Into Eyes – Immediately rinse eyeball and inner surface of eyelid with water for 15 minutes by forcibly holding the eye open. Seek medical attention. Notify supervisor and EH&S at x59797 immediately. Medical Emergency Dial 911 or x52111 Life Threatening Emergency, After Hours, Weekends And Holidays – Dial 911 (or 310825-1491 from cell phone) or contact the Ronald Reagan UCLA Medical Center (emergency room) directly at x52111 (located at 757 Westwood Plaza, enter from Gayley Avenue). Note: All serious injuries must be reported to EH&S at x59797 within 8 hours. Non-Life Threatening Emergency– Go to the Occupational Health Facility (OHF), x56771, CHS room 67-120 (This is on the 6th floor, 7th corridor, room 120. Enter through the School of Dentistry on Tiverton Drive and proceed to the “O” elevator to the 6th floor.)Hours: M - F, 7:30 a.m. to 4:30 p.m. At all other times report to Ronald Regan UCLA Medical Center (emergency room) at x52111. Note: All serious injuries must be reported to EH&S at x59797 within 8 hours. Needle stick/puncture exposure (as applicable to chemical handling procedure)– Wash the affected area with antiseptic soap and warm water for 15 minutes. For mucous membrane exposure, flush the affected area for 15 minutes using an eyewash station. Page the needle stick nurse by dialing 231 from a campus phone, enter 93333 when prompted and then enter your extension. Hours: M – F, 8:00 a.m. to 4:00 p.m. At all other times report to Ronald Regan UCLA Medical Center (emergency room) at x52111. Note: All needle stick/puncture exposures must be reported to EH&S at x59797 within 8 hours. Decontamination/Waste Disposal Procedure Clean areas where BrdU has been handled by adding water, followed with a soap and water wash. No waste streams containing BrdU shall be disposed of in sinks or general refuse. Extra BrdU is considered as hazardous chemical waste and disposed of in accordance with UCLA hazardous waste protocol. Absorbent materials (e.g., associated with spill cleanup) grossly contaminated shall be handled in accordance with the chemical waste disposal system. Material Safety Data Sheet (MSDS) Location (State the location of MSDS) Hardcopy or electronic copy must be available. Online MSDS can be accessed at http://msds.ehs.ucla.edu. Protocol/Procedure (Add specific description of procedure) Note: Any deviation from this SOP requires written approval from PI. 4 Documentation of Training (signature of all users is required) Prior to conducting any work with Bromo-deoxyuridine, designated personnel must provide training to his/her laboratory personnel specific to the hazards involved in working with this substance, work area decontamination, and emergency procedures. The Principal Investigator must provide his/her laboratory personnel with a copy of this SOP and a copy of the Bromo-deoxyuridine MSDS provided by the manufacturer. The Principal Investigator must ensure that his/her laboratory personnel have attended appropriate laboratory safety training or refresher training within the last two years. I have read and understand the content of this SOP: Name Signature Date ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ ________________________________ ____________________________ ____________ 5