Use of DMBA in Animals

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SOP #: 002 VERSION NO.: 1.0 PREPARED BY: Catherine Brennan (EHS) SUPERCEDES: n/a EFFECTIVE DATE: August 2010 LAST REVISED: April 5, 2018 Standard Operating Procedure For Use of DMBA in Animals PURPOSE: This document establishes procedures for the safe handling and use of 7,12- dimethylbenz(a)anthracene, commonly known as DMBA (CAS# 57-97-6) in university Division of Laboratory Animal Medicine (DLAM) facilities. DMBA is a highly potent carcinogen that causes cancer and heritable genetic damage and is used to induce cancer in animal models. Contact with eyes, skin and clothing should be avoided and inhalation of dust must be prevented. Based on the risk associated with the use of DMBA the safety procedures outlined below are required by all research staff and DLAM employees when working with DMBA in DLAM facilities. REQUIREMENTS: Animal Housing: All animal work associated with this chemical, will be conducted in the ABSL2 isolation cubicles. A Chemical Hazard sign should be posted on the entry door to indicate DMBA use. The IACUC Chemical Hazard form should be posted on cubicle or entry door to indicate DMBA use. Individual cages should be marked with a yellow chemical hazard card. Cages where DMBA is administered to the animals should not be changed for 48 hours after last dose. If cages are in need to be changed research personnel will change the animal bedding. Administrative Controls: The IACUC Use of Chemical Hazards in Animals form should be submitted and approved by EHS prior to working with DMBA in animals. Anyone who handles DMBA or works with animals that have been treated with DMBA is required to review this SOP and the attached Material Safety Data Sheet (MSDS) prior to work.

Engineering Controls: Procedures that involve handling DMBA must be conducted under exhaust ventilation in a chemical fume hood or ducted Biological Safety Cabinet (BSC). Cage changing occurs within a Biological Safety Cabinet (BSC). Personal Protective Equipment (PPE): Standard entry, gowning, and personal hygiene procedures of ABSL2 will be followed. Wear double nitrile gloves when preparing or dispensing DMBA solutions Safety glasses will be worn when handling DMBA, in the animal holding room and when cage changing as an additional precaution. Waste Disposal: All animal carcasses are to be incinerated. Cages are to be autoclaved as per standard ABSL2 procedures. Animal bedding is to be incinerated during dosing and for 48 hours after last dose. Unused solutions of DMBA and contaminated solid waste will be disposed of as hazardous material (https://ehs.cloudapps.unc.edu/hazmat_pickup/) Accidents or Injuries: Animal bites do not pose any increased risk. As per standard policy, report to supervisor. If DMBA solution is splashed on an individual or in eyes flush for 15 minutes with copious quantities of water and contact the University Employee Occupational Health Clinic (966-9119). Spill procedures: o Do not attempt to clean-up if you feel unsure of your ability to do so or if you perceive the risk to be greater than normal laboratory operations. o If a small spill occurs absorb with absorbent pads. Double bag contaminated solid waste and submit for hazardous material disposal (https://ehs.cloudapps.unc.edu/hazmat_pickup/). o If a large spill occurs notify others in the area and evacuate room immediately. Contact EHS (962-5507) during working hours and 911 if after hours. 2

SIGMA-ALDRICH sigma-aldrich.com Material Safety Data Sheet Version 4.0 Revision Date 03/13/2010 Print Date 08/06/2010 1. PRODUCT AND COMPANY IDENTIFICATION Product name : Product Number : D3254 Brand : Sigma Company : Sigma-Aldrich 3050 Spruce Street SAINT LOUIS MO 63103 USA Telephone : +18003255832 Fax : +18003255052 Emergency Phone # : (314) 776-6555 2. HAZARDS IDENTIFICATION Emergency Overview OSHA Hazards Toxic by ingestion, Carcinogen, Teratogen, Mutagen GHS Label elements, including precautionary statements Pictogram Signal word Hazard statement(s) H302 H316 H350 Danger Harmful if swallowed. Causes mild skin irritation. May cause cancer. Precautionary statement(s) P201 Obtain special instructions before use. P308 + P313 IF exposed or concerned: Get medical advice/attention. HMIS Classification Health hazard: 2 Chronic Health Hazard: * Flammability: 0 Physical hazards: 0 NFPA Rating Health hazard: 2 Fire: 0 Reactivity Hazard: 0 Potential Health Effects Inhalation May be harmful if inhaled. May cause respiratory tract irritation. Skin May be harmful if absorbed through skin. May cause skin irritation. Eyes May cause eye irritation. Ingestion Toxic if swallowed. 3. COMPOSITION/INFORMATION ON INGREDIENTS Synonyms : 1,4-Dimethyl-2,3-benzophenanthrene Sigma - D3254 Page 1 of 6

Formula : C 20 H 16 Molecular Weight : 256.34 g/mol DMBA 9,10-Dimethyl-1,2-benzanthracene CAS-No. EC-No. Index-No. Concentration 57-97-6 200-359-5 - - 4. FIRST AID MEASURES General advice Consult a physician. Show this safety data sheet to the doctor in attendance.move out of dangerous area. If inhaled If breathed in, move person into fresh air. If not breathing give artificial respiration Consult a physician. In case of skin contact Wash off with soap and plenty of water. Consult a physician. In case of eye contact Rinse thoroughly with plenty of water for at least 15 minutes and consult a physician. If swallowed Never give anything by mouth to an unconscious person. Rinse mouth with water. Consult a physician. 5. FIRE-FIGHTING MEASURES Suitable extinguishing media Use water spray, alcohol-resistant foam, dry chemical or carbon dioxide. Special protective equipment for fire-fighters Wear self contained breathing apparatus for fire fighting if necessary. 6. ACCIDENTAL RELEASE MEASURES Personal precautions Use personal protective equipment. Avoid dust formation. Avoid breathing dust. Ensure adequate ventilation. Evacuate personnel to safe areas. Environmental precautions Prevent further leakage or spillage if safe to do so. Do not let product enter drains. Methods and materials for containment and cleaning up Pick up and arrange disposal without creating dust. Keep in suitable, closed containers for disposal. 7. HANDLING AND STORAGE Precautions for safe handling Avoid formation of dust and aerosols. Provide appropriate exhaust ventilation at places where dust is formed. Normal measures for preventive fire protection. Conditions for safe storage Keep container tightly closed in a dry and well-ventilated place. Keep in a dry place. Sigma - D3254 Page 2 of 6

8. EXPOSURE CONTROLS/PERSONAL PROTECTION Contains no substances with occupational exposure limit values. Personal protective equipment Respiratory protection Where risk assessment shows air-purifying respirators are appropriate use a full-face particle respirator type N100 (US) or type P3 (EN 143) respirator cartridges as a backup to engineering controls. If the respirator is the sole means of protection, use a full-face supplied air respirator. Use respirators and components tested and approved under appropriate government standards such as NIOSH (US) or CEN (EU). Hand protection Handle with gloves. Eye protection Face shield and safety glasses Skin and body protection Choose body protection according to the amount and concentration of the dangerous substance at the work place. Hygiene measures Avoid contact with skin, eyes and clothing. Wash hands before breaks and immediately after handling the product. 9. PHYSICAL AND CHEMICAL PROPERTIES Appearance Form powder Colour Safety data ph Melting point Boiling point Flash point Ignition temperature Lower explosion limit Upper explosion limit Water solubility light yellow 122-123 C (252-253 F) - lit. 10. STABILITY AND REACTIVITY Chemical stability Stable under recommended storage conditions. Conditions to avoid Materials to avoid Strong oxidizing agents Hazardous decomposition products Hazardous decomposition products formed under fire conditions. - Carbon oxides 11. TOXICOLOGICAL INFORMATION Acute toxicity LD50 Oral - rat - 327 mg/kg Skin corrosion/irritation Skin - mouse - Mild skin irritation Sigma - D3254 Page 3 of 6

Serious eye damage/eye irritation Respiratory or skin sensitization Germ cell mutagenicity Laboratory experiments have shown mutagenic effects. Carcinogenicity This product is or contains a component that has been reported to be probably carcinogenic based on its IARC, OSHA, ACGIH, NTP, or EPA classification. Possible human carcinogen IARC: No component of this product present at levels greater than or equal to 0.1% is identified as probable, possible or confirmed human carcinogen by IARC. ACGIH: No component of this product present at levels greater than or equal to 0.1% is identified as a carcinogen or potential carcinogen by ACGIH. NTP: No component of this product present at levels greater than or equal to 0.1% is identified as a known or anticipated carcinogen by NTP. OSHA: No component of this product present at levels greater than or equal to 0.1% is identified as a carcinogen or potential carcinogen by OSHA. Reproductive toxicity Laboratory experiments have shown teratogenic effects. Specific target organ toxicity - single exposure (GHS) Specific target organ toxicity - repeated exposure (GHS) Aspiration hazard Potential health effects Inhalation May be harmful if inhaled. May cause respiratory tract irritation. Ingestion Toxic if swallowed. Skin May be harmful if absorbed through skin. May cause skin irritation. Eyes May cause eye irritation. Additional Information RTECS: CW3850000 12. ECOLOGICAL INFORMATION Toxicity Persistence and degradability Bioaccumulative potential Mobility in soil PBT and vpvb assessment Other adverse effects Sigma - D3254 Page 4 of 6

13. DISPOSAL CONSIDERATIONS Product Observe all federal, state, and local environmental regulations. Contact a licensed professional waste disposal service to dispose of this material. Dissolve or mix the material with a combustible solvent and burn in a chemical incinerator equipped with an afterburner and scrubber. Contaminated packaging Dispose of as unused product. 14. TRANSPORT INFORMATION DOT (US) UN-Number: 3077 Class: 9 Packing group: III Proper shipping name: Environmentally hazardous substances, solid, n.o.s. Reportable Quantity (RQ): 1 lbs Marine pollutant: No Poison Inhalation Hazard: No IMDG Not dangerous goods IATA Not dangerous goods 15. REGULATORY INFORMATION OSHA Hazards Toxic by ingestion, Carcinogen, Teratogen, Mutagen DSL Status All components of this product are on the Canadian DSL list. SARA 302 Components SARA 302: No chemicals in this material are subject to the reporting requirements of SARA Title III, Section 302. SARA 313 Components SARA 313: This material does not contain any chemical components with known CAS numbers that exceed the threshold (De Minimis) reporting levels established by SARA Title III, Section 313. SARA 311/312 Hazards Acute Health Hazard, Chronic Health Hazard Massachusetts Right To Know Components Pennsylvania Right To Know Components New Jersey Right To Know Components California Prop. 65 Components WARNING! This product contains a chemical known to the State of California to cause cancer. CAS-No. 57-97-6 CAS-No. 57-97-6 CAS-No. 57-97-6 CAS-No. 57-97-6 Revision Date 1993-04-24 Revision Date 1993-04-24 Revision Date 1993-04-24 Revision Date 2007-09-28 16. OTHER INFORMATION Further information Copyright 2010 Sigma-Aldrich Co. License granted to make unlimited paper copies for internal use only. Sigma - D3254 Page 5 of 6

The above information is believed to be correct but does not purport to be all inclusive and shall be used only as a guide. The information in this document is based on the present state of our knowledge and is applicable to the product with regard to appropriate safety precautions. It does not represent any guarantee of the properties of the product. Sigma-Aldrich Co., shall not be held liable for any damage resulting from handling or from contact with the above product. See reverse side of invoice or packing slip for additional terms and conditions of sale. Sigma - D3254 Page 6 of 6