Standard Operating Procedure Section 1: Laboratory Information Principal Investigator Name: Position Title: Department: Email Address: Primary After Hours Phone: WENBIN LIN PROFESSOR CHEMISTRY wenbinlin@uchicago.edu 919-357-0969 Safety Supervisor (person responsible for safety in absence of Principal Investigator) Name: CHRISTOPHER POON Lab Phone: 773-702-3605 Email Address: chrisp22@uchicago.edu Primary After Hours Phone: 206-852-1657 Name: Lab Phone: Email Address: Primary After Hours Phone: MAREK PIECHOWICZ 773-702-3605 mpiechowicz@uchicago.edu 847-363-1393 Locations Building: Gordon Center for Integrated Science Room E506 E513 E547B E547C E547D E547E E547H Special Notes Hazardous Chemicals & Equipment, Biohazards, BSL-2 Hazardous Chemicals & Equipment Hazardous Chemicals & Equipment, Shipping Hazardous Chemicals & Equipment Hazardous Chemicals & Equipment Hazardous Chemicals & Equipment Hazardous Chemicals & Equipment Section 2: Safety Training Every lab personnel is required by Environmental Health and Safety to have Laboratory Safety Training before being allowed to work in the laboratory. This can be done here: https://safety.uchicago.edu/training/index.shtml. Click “Determine What Training You Need” to find out the required trainings. Note: You should only register as a laboratory worker (not a radiation worker). If you are doing x-ray crystallography, do x-ray safety and awareness training. If you are in the nanomedicine group, please do the biological safety cabinets training. The fire safety and evacuation training must be done annually. The chemical hygiene plan must be done every three years. Location and availability of the chemical hygiene plan (Lab safety manual and lab safety plan) can be found online: http://safety.uchicago.edu/pp/labsafety/hygiene.shtml Location and availability of known reference material, including MSDS, on the hazards, safe handling, storage, and disposal of hazardous chemicals can be found online: http://safety.uchicago.edu/pp/labsafety/hazmat.shtml Physical hazards and health hazards of chemicals in the workplace are located in the online inventory located on the group webpage. Section 3: Personal Protective Equipment and Safety: All personnel must be trained in the procedures described here. PPE required: Goggles Gloves Lab coat: See specific material for details. ALWAYS have someone else present in the lab when using these materials! Laboratory Operation CHEMICALS: VOLUMES < 1 LITER PER CONTAINER Hazard SKIN CONTACT; EYE AND FACE EXPOSURE CHEMICALS: VOLUMES > 1 LITER PER CONTAINER SKIN CONTACT; EYE AND FACE EXPOSURE HIGHLY TOXIC OR CARCINOGENIC CHEMICALS SKIN EXPOSURE WORKING WITH GLASSWARE UNDER REDUCED or ELEVATED PRESSURES HANDLING HOT or COLD MATERIALS HANDLING BROKEN GLASS; INSERTING GLASS TUBES INTO GLASS STOPPERS AND HANDLING OTHER SHARP EDGED OBJECTS EXPLOSION HAZARDS CHEMICAL SPRAY TO FACE NANOMATERIALS SKIN AND INHALATION (CURRENTLY LIMITED HAZARD DATA IS KNOWN) SKIN BURNS CUTS INJURY FROM FLYING PROJECTILES PPE Required GLOVES (CONSULT GLOVE PERMEABILITY CHARTS)(SPECIFY TYPE), SAFETY GLASSES WITH SIDE SHIELDS, LAB COATS GLOVES (CONSULT GLOVE PERMEABILITY CHARTS)(SPECIFY TYPE), SAFETY GLASSES WITH SIDE SHIELDS, LAB COATS, RUBBER APRONS (CORROSIVES) GLOVES (DOUBLE GLOVING RECOMMENDED), LAB COATS, DISPOSABLE JUMPSUITS, SAFETY GLASSES WITH SIDE SHIELDS CHEMICAL SPLASH GOGGLES, GLOVES, LAB COATS INSULATED GLOVES, e.g. NOMEX or KEVLAR LEATHER GLOVES FACE SHIELD, CHEMICAL SPLASH GOGGLES, LAB COAT, GLOVES GLOVES (DOUBLE GLOVE OR CHANGE GLOVES FREQUENTLY), EYE PROTECTION, LABCOATS, DISPOSABLE JUMPSUITS, RESPIRATOR WITH HEPA CYTOTOXIC / ANTINEOPLASTIC AGENTS ADVERSE HEALTH EFFECTS DUE TO OCCUPATIONAL EXPOSURE FILTER IF EXPOSED TO NANOAEROSOLS NITRILE GLOVES (DOUBLE GLOVE), EYE PROTECTION, LAB COATS OR DISPOSABLE JUMPSUITS Either work in a fume hood or wear a face shield. While goggles are important, they do not protect your entire face from possible contact with pyrophoric materials. Care must be taken when working with pyrophoric materials in a fume hood. If a fire does occur, DO NOT shut the hood sash as this will intensify the fire by increasing the air flow and thus provide more oxygen for the fire. Wear appropriate clothing. Synthetic fibers should be avoided as they are highly flammable and can exacerbate the hazards. Wear closed-toe shoes and tie back loose hair to prevent ignition in the event of a flash fire. Section 4: Handling of Hazardous Chemicals: All toxic chemicals/laboratory gloves must not be brought into the designated food areas within the laboratory. In addition, PPE must be worn while doing bench work. All laboratory workers may handle carcinogens and are expected to educate themselves, or ask the laboratory safety manager for assistance in finding the MSDS. Handling air/water sensitive reagents: Reactions involving our air-sensitive reagents can be carried out in common ground-glass apparatus. Other equipment required are a source of inert gas, a septum inlet, a bubbler, and syringes fitted with suitable needles. Laboratory glassware contains a thin film of adsorbed moisture which can be easily removed by heating in an oven (125°/overnight or 140°/4 hrs). The hot glassware should be cooled in an inert atmosphere by assembling the glassware while hot and flushing with a stream of dry nitrogen or argon. A thin film of silicone or hydrocarbon grease must be used on all standard-taper joints to prevent seizure upon cooling. Alternatively, the apparatus may be assembled cold and then warmed with a heat gun while flushing with dry nitrogen. Only high-purity, dry nitrogen from a cylinder with a pressure regulator (adjusted to 3-5 psi) should be used for flushing. Plastic tubing can be used to connect the nitrogen line to a tube connector adapter (equipped with a stopcock) on the reaction apparatus. Nitrogen may also be introduced through a rubber septum via a hypodermic needle connected to the end of the flexible tubing on the nitrogen line. The needle-tubing connector provides a simple method for attaching the needle to the tubing. When not in use, this nitrogen-flushing needle should be closed by inserting the needle into a solid rubber stopper to prevent diffusion of air into the needle when the nitrogen is turned off. Small quantities (up to 50 mL) of air-sensitive reagents and dry solvents may be transferred with a syringe equipped with a 1-2ft long needle. These needles are used to avoid having to tip reagent bottles and storage flasks. Tipping often causes the liquid to come in contact with the septum causing swelling and deterioration of the septa, and should therefore be avoided. A rubber septum provides a positive seal for only a limited number of punctures depending on needle size. Therefore, always reinsert the needle through the existing hole. It is also advantageous to put a layer of silicone or hydrocarbon grease on a rubber septum to facilitate passage of the needle through the rubber and to minimize the size of the hole in the septum. Ideally, the syringe and needle should be dried in an oven prior to use. Naturally, the syringe body and plunger should not be assembled before being placed in the oven. The syringe should be flushed with nitrogen during the cooling. A syringe may also be flushed 10 or more times with dry nitrogen to remove the air and most of the water adsorbed on the glass. A dry syringe may be closed to the atmosphere by inserting the tip of the needle into a rubber stopper. To accomplish the double-tipped needle transfer, the needle is first flushed with nitrogen. The Sure/Seal bottle is pressurized with nitrogen using the nitrogen flushing needle. The double tipped needle is then inserted through the septum on the reagent bottle into the head space above the reagent. Nitrogen immediately passes through the needle. Finally, the other end of the double-tipped needle is inserted through the septum on the reaction apparatus, and the end of the needle in the reagent bottle is pushed down into the liquid. The volume of liquid reagent transferred is measured by using a calibrated flask or addition funnel. When the desired volume has been transferred, the needle is immediately withdrawn to the head space above the liquid, flushed slightly with nitrogen, and removed. The needle is first removed from the reaction apparatus and then from the reagent bottle. Handling of Distillation Stills and/or Solvent System: Prior to usage of distillation stills, laboratory workers must be trained in appropriate usage. It is the responsibility of the person who set the still up and the laboratory safety manager to make sure that the still is operating correctly and does not pose an explosion hazard. The PI/lab manager will designate a member of the lab to maintain the solvent stills. Handling of Base Bath: Prior to handling base bath, lab coat, safety glasses, and neoprene gloves (with latex or nitrile gloves underneath) must be worn at all time. Close-toed shoes are required as drops of base bath lead to holes in clothing. The base bath must be covered at all time with a secondary containment underneath to prevent spills. To make a new base bath, potassium hydroxide (500g) should be first dissolved in 1L water in a beaker. Beware that the solution will become very hot upon dissolution. This solution should be added to 12L of isopropanol in a large Nalgene container. Disposal of base bath should be handled by removing from its plastic container, either by siphoning into a new waste container, or removal 1 L at a time using a large beaker. The waste container should only contain caustic solution and should not be mixed with other waste. Waste containers should not be more than 95% full and cannot stay in the lab for more than 6 months at which the waste container is started. The waste containers must be marked to identify the contents, hazards, and accumulation start and end dates for EHS to pickup. In case of large exposure, the victim should be removed from the contaminated area, placed under a safety shower and/or eyewashes while emergency personal are contacted. If large spill occurred, evacuate the area and EHS should be contacted. For smaller spill, the liquid can be absorbed by dry sand. Handling of Hydrofluoric Acid (HF): When handling hydrofluoric acid, chemical splash goggles, face shield, chemical splash apron, medium or heavy weight viton, nitrole, or natural rubber gloves must be worn at all time. A second pair of nitrile exam gloves should be worn under the gloves for protection against leaks. If HF is spilled on glove, removed contaminated gloves, wash your hands thoroughly, and check hands for any sign of contamination. Since HF is corrosive and will rapidly damages tissues, an eyewash and shower must be nearby for accessibility. Use of HF must be done inside a fume hood and should never be use while working alone after hours. A designated are for HF with warning sign is located in the middle room closest to the safety shower. HF must be used in polyethylene, polypropylene, Teflon, wax, lead, or platinum containers. HF should never be use in glass, ceramic, and metal container. Disposal of HF should be placed in a polyethylene or Teflon container with a sealed lid and labels. Waste containers should not be more than 95% full and cannot stay in the lab for more than 6 months at which the waste container is started. The waste containers must be marked to identify the contents, hazards, and accumulation start and end dates for EHS to pickup. In the case of spill, if more than 1 L of HF is spilled outside of the chemical hood, evacuate the area, close the doors, and post the area with sign to prevent others from entering. Notify University Police at 123 or 773-702-8181. If spill is less than 1 L inside a chemical fume hood, neutralize the spill with Spill-X-C caustic neutralizer (in the hallway), caustic soda, powdered calcium carbonate, calcium hydroxide, or a commercial HF spill kit. Since HF can produce delayed effects and serious tissue damage without producing pain, any exposure must receive first aid and medical evaluation. Flush affected area for at least 15 minutes with an emergency shower. Remove all contaminated clothing. Apply calcium gluconate gel to the affected area as soon as possible and contact University Police. Those assisting the victims must wear proper PPE to prevent contamination themselves. If exposure occurs in eyes, flush eyes while holding eyelids opened for at least 15 minutes. If HF is ingested, dilute acid by giving large amount of water. Drink several glasses of milk or take 8-12 tablets of Tums or Rolaids. Do not induce vomiting and never give anything by mouth to an unconscious or convulsing person. Handling of N,N-Dimethylformamide (DMF): Safety glasses, gloves, lab coat, long pants, and close-toed shoes must be worn when handling DMF. Neither latex nor nitrile gloves would provide adequate protection against DMF. If DMF is spilled onto a glove, it should be removed immediately and placed into proper waste container, followed by washing of hands with soap and water. Storage of DMF should be placed in flammable-safe, well ventilated areas and be used in a chemical fume hood. DMF waste should be disposed in hazardous organic waste. If more than 1 L of DMF is spilled, call University Police. If the spill is less than 1 L and in a fume hood, DMF spill can be cleaned up. Section 5: Chemical Storage and Segregation: STORAGE GROUP * Compressed Gases - Flammable Compressed Gases - Oxidizing Compressed Gases – Poisonous Corrosives – Acids INORGANIC Corrosives – Acids ORGANIC Corrosives – Bases Explosives Flammable Liquids Flammable Solids RECOMMENDED STORAGE METHOD Store in a cool, dry area, away from oxidizing gases. Securely strap or chain cylinders to a wall or bench top. Store in a cool, dry area, away from flammable gases and liquids. Securely strap or chain cylinders to a wall or bench top. Store in a cool, dry area, away from flammable gases and liquids. Securely strap or chain cylinders to a wall or bench top. Store in a separate, lined/protected acid storage cabinet. *DO NOT store acids on metal shelves* Store in a separate, lined/protected acid storage cabinet. *DO NOT store acids on metal shelves* Store in a separate storage cabinet. Store in a secure location away from all other chemicals. Do not store in an area where they can fall. Store in a flammable storage cabinet. *Peroxide forming chemicals must be dated upon opening e.g. Ether, Tetrohydrofuran * Store in a separate dry cool area away from oxidizers, corrosives. CHEMICAL EXAMPLES Methane, Acetylene, Propane INCOMPATIBLES SEE MSDS IN ALL CASES Oxidizing and toxic compressed gases, oxidizing solids. Oxygen, Chlorine, Bromine Flammable gases. Carbon monoxide, Hydrogen sulfide Flammable and/or oxidizing gases. Inorganic (mineral) acids Hydrochloric acid, Sulfuric acid, Chromic acid, Nitric acid. Organic acids - Acetic acid, Trichloroacetic acid, Lactic acid Flammable liquids, flammable solids, bases, and oxidizers. Organic acids Flammable liquids, flammable solids, bases, and oxidizers. Inorganic acids Flammable liquids, oxidizers, poisons, and acids. All other chemicals. Ammonium hydroxide, Potassium hydroxide, Sodium hydroxide Ammonium Nitrate, Nitro Urea, Sodium azide, Trinitroaniline, Trinitroanisole, Trinitrobenzene, Trinitrophenol/Picric acid, Trinitrotoluene (TNT). Acetone, Benzene, Diethyl ether, Methanol, Ethanol, Hexanes, Toluene Phosphorus, Carbon, Charcoal Acids, bases, oxidizers, and poisons. Acids, bases, oxidizers, and poisons. Water Reactive Chemicals Oxidizers Poisons/Toxic General Chemicals -Non-Reactive Store in a dry, cool location. Protect from water and the fire sprinkler system, if applicable. Label location - WATER REACTIVE CHEMICALS Store in a spill tray inside a noncombustible cabinet, separate from flammable and combustible materials. Sodium metal, Potassium metal, Lithium metal, Lithium Aluminium hydride Separate from all aqueous solutions, and oxidizers. Sodium hypochlorite, Benzoyl peroxide, Potassium permanganate, Potassium chlorate, Potassium dichromate. The following are generally considered oxidizing substances: Peroxides, Perchlorates, Chlorates, Nitrates, Bromates, Superoxides Separate from reducing agents, flammables, and combustibles and organic materials. Store separately in a vented, cool, dry, area in chemically resistant secondary containers. Store on general laboratory benches or shelving. Cyanides, heavy metal compounds, i.e. Cadmium, Mercury, Osmium Agar, Sodium chloride, Sodium bicarbonate, and most nonreactive salts Flammable liquids, acids, bases, and oxidizers. See MSD Section 6: Hazardous Waste Disposal: Liquid and solid hazardous waste: Processes which create chemical waste include, but are not limited to centrifuging, running columns, microemulsions, reactions, and rotovapping. We expect to generate ignitable, corrosive, and toxic waste. Waste streams generated are broken down into non-halogenated solvents, halogenated solvents, acids, and bases. We have multiple waste bottles designated for specific waste types with the type labeled on the front of the bottle. Centralized waste containers are located under the rotovap in GCIS E547C and GCIS E513. Incompatible waste bottles are stored in different sections of the lab. When containers are full, they are submitted for pickup weekly. Any questions about chemical storage are referred to the lab safety manager. Disposal of concentrated nitric acid should be done by diluting the acid to 2% with water (adding acid to water, not the other way around). The diluted nitric acid can be disposed in the 2% nitric acid waste located under the ICP-MS. Do not tighten the lid as it could cause pressure buildup. If the nitric acid contains any mice tissue, please dispose the acid in Hood L inside the plastic container labeled “Concentrated nitric acid waste with mice tissue” with a vented cap. When container is more than 50% full, please replace the cap with the normal cap next to the waste container and do not completely tighten the lid. Notify the safety officer for disposal. Clean the vented cap thoroughly for reusable purpose. Sharps: Collect in one gallon red containers bearing biohazard symbol are located in each section of the lab. NEVER fill beyond 3/4 full. Filled containers are put in large red bin for pickup. Call (4-2761) for pickup. Broken Glass: Any glass wastes are placed in glass waste bin located in each section of the lab. When full, they are taped up tightly. Call (4-1414) for pickup. Section 7: Emergency Procedures: Procedures for Hazardous Chemicals: Chemical Spills: If any chemicals are spilled within the lab in large quantities, the spill kit, located in GCIS E547C, will be used to neutralize and contain the spill. Alert your neighbors. All nonessential lab workers will be cleared out of the laboratory, any fume hoods/heat sources will be turned off and closed. If necessary, first aid or decontamination procedures will be used. If severe, use safety shower or eyewash and remove affected clothing and rinse for a minimum of 15 minutes. Shut doors to the spill area and evacuate the area, if necessary. Report the incident to your lab director, principal investigator, or department administer. Contact EHS at (2-9999) for advice or help, if needed. Follow steps in Lab Safety Manual or in spill kit. Acid or base spills: If a strong acid or base is spilled, sodium bicarbonate will be used to neutralize the solution and absorb any liquids. Handling Hazardous Chemicals: PI/lab safety manager will be sure that workers are sufficiently trained when using overly hazardous chemicals and will supervise novice workers. Medical Incident: Assist the victim, if possible. Report the incident to your lab director, principal investigator, or department administrator. Fill out the Student Injury /Illness Assessment Form. Report incident to EHS at (2-9999). If emergency, call UC Police at 123 (on-campus phone) or 773-702-8181 (off-campus phone). Fire, Flammable gas, or Nature gas: Evacuate the area, if necessary. Shut doors to the area and alert others. Pull the nearest fire alarm and call UC Police at 911. If minor fire or non-life threatening situation, use extinguisher as appropriate. Report incident to your lab director, principal investigator, or department administrator. Report incident to EHS at (2-9999). Section 8: Biological Hazards: Human/primate source material, BSL-2, is designated based on the assumption each worker has normal, healthy adult immune system. Some BSL-2 materials stored in GCIS E506 are continuous human cell lines and HELA human cells. Universal/Standard precautions are taken to prevent transmission of human immunodeficiency virus(HIV), hepatitis B virus(HBV), and other bloodborne pathogens. All workers of this material will be enrolled in Bloodborne Pathogens program as mandated by OSHA. In addition, in order to work in the cell lab, a biological safety cabinet (BSC) training must be done. Trainings can be found at https://safety.uchicago.edu/training/index.shtml. Approved products for surface decontamination in BSC include sodium hypochlorite (bleach solution) at 6.15% for 2 minutes and 70 % ethanol 190 or 200 proof for 1 minute. Frequent disinfectant is done prior and after usage of BSC. Disinfection products are located near the BSC. Potentially hazardous laboratory procedures include centrifugation using proper aseptic technique, syringes needles, or other sharps in a biological safety cabinet, pipetting in a biological safety cabinet, opening culture plates, tubes, bottles, or ampoules in a biological safety cabinet. Biohazard waste generated includes sharps, plastic/glass pipettes; pipette tips, solids, liquid. Biohazard liquid waste generated includes cell medium, phosphate buffer saline and other biological assays. Chemically disinfected liquid wastes are not used for propagating microbes/viral vector/toxins. Emergency Procedure: Spill Inside BSC: 1. Contain spill with absorbent paper. 2. Dampen Paper with disinfectant. Allow to stand for 20 minutes. 3. If sharps/glass is present, use mechanical means to collect the waste (eg. forceps, cardboard flaps). 4. Remove gloves after area is decontaminated. 5. Wash hands. Large Spill Inside BSC: 1. If splash has occurred outside the cabinet resulting in personnel exposure to infectious material, the Principal Investigator and EHS should be notified and the need for prophylactic treatment or other medical attention determined. 2. Contaminated clothing should be removed and containerized for autoclaving. 3. Thoroughly wash hands and face, if exposure has occurred. 4. Remove gloves after area is decontaminated 5. Chemical decontamination procedures should be initiated at once while the cabinet continues to operate to prevent escape of contaminants from the cabinet. 6. Spray or wipe walls, work surfaces, and equipment with appropriate disinfectant. 7. Flood top tray, drain pans, and catch basin below work surfaces with disinfectant and allow to stand 20 minutes. 8. Dump excess disinfectant from tray and drain pans into cabinet base. 9. Lift out tray and removable exhaust grille work. Wipe off top and bottom (underside) surfaces with disinfectant sponge or cloth. Replace in position. 10. Gloves, cloth or sponge should be discarded in an autoclave pan and autoclaved. 11. Drain disinfectant from cabinet base into an appropriate container and autoclave. 12. Remove gloves and wash hands. 13. This procedure does not decontaminate the interior parts of the cabinet such as the filters, blowers, and air ducts. If the entire cabinet is to be decontaminated with formaldehyde gas, contact EHS (2-9999). Spill Outside BSC: 1. Remove contaminated outer gloves. 2. Notify others in lab of spill. 3. Decontaminate all personnel, clothing and exit laboratory 4. Notify PI and EHS. 5. Allow aerosols to settle for 30 minutes. 6. Re-enter wearing PPE (including respiratory protection). 7. Carefully dilute spill from outside edges using appropriate disinfectant. 8. Allow 20 minutes contact time. 9. Clean-up with paper towels. 10. Decontaminate clean-up materials. 11. Wash hands thoroughly. Biohazard Waste Disposal Procedures Sharps: Collect in one gallon red cans bearing biohazard symbol and 'x' with indicator tape. NEVER fill beyond 3/4 full. Plastic and/or glass pipettes: Puncture resistant outer container bearing the biohazard symbol (outer box 'x' with indicator tape) lined with orange or red autoclave bag ('x' the biohazard symbol on bag before loading the bag) Pipettes tips: Small bench top biohazard bag with indicator tape 'x' Solid: Hardwalled biohazard waste collection container lined with orange autoclave bag bearing indicator tape 'x' over biohazard symbol Filled biohazard wastes are put inside large red bins to be pickup for autoclave. Call (4-2761) for pickup. Section 9: Personnel Consent Name Carter Abney Levi Bennish Dean Boures Christina Chan Alex Chen Michaël Carboni Xiaopin Duan Chunbai He Connor Gilhula Frank Greene Nining Guo Pengfei Ji Zekai Lin Demin Liu Jianqin Lu Kuangda Lu Kristine Ma Kuntal Manna Lexie McIsaac Dan Micheroni Alexei Owen Marek Piechowicz Christopher Poon Takahiro Sawano Nathan Thacker Ania Urban Marcela Wanderley Youfu Wang Ruoyu Xu Teng Zhang Xin Zhou Linus Zhu Initial CWA LYB DB CC AC MC XD CH JCG Date 02May2014 7/13/2015 2/22/15 5/8/2014 5/2/14 5/2/14 05/05/2014 05-02-2014 5-4-14 F.G. 5/2/2014 2015/8/3 PF J. ZL DL JL KL KM KM LM DM AO MP CP TS NT August 3rd, 2014 5/8/2014 05/08/2014 05/08/2015 05/02/2014 5/8/14 07/23/2015 5/5/14 08/03/2015 04.12.15 06.16.2014 5/2/14 05/02/2014 5/3/2014 M.W. YFW R.X. T.Z. X LZ 05.02.2014 07/28/2015 01/02/2015 05.08.2014 May.2nd, 2014 06/25/2015