SAFETY MEETING LEIGHTON LABORATORY Prepared by Archana Sudame

SAFETY MEETING LEIGHTON LABORATORY -Prepared by Archana Sudame

OSHA REQUIREMENTS SAFETY TRAINING: You must receive adequate training in working with hazardous chemicals. CHP: You must have access to your laboratory’s Chemical Hygiene Plan. MSDS: Material Safety Data Sheets must be made available to you. CHEMICAL INVENTORY: You must be informed of hazardous chemicals present in your laboratory and of operations in which they are involved.

Injury and Illness Prevention Program (IIPP) An Injury and Illness Prevention Program (IIPP) is a department's ongoing program for preventing work-related injuries and illnesses and for creating a safe and healthful work environment Refer to the Yellow Flip Chart of CHP

IIPP specifies that • there will be initial training for new employees and when the IIPP was first established • there will be training upon changes in the working environment o with new job assignments o whenever new substances, processes, procedures, or equipment are introduced o whenever the employer is made aware of new or previously unrecognized hazards

Hazard Communication Do you know: • Where to find information concerning the hazards of the chemicals present in your work area? • When to use a laboratory chemical hood? • How to select appropriate eye protection? • What about gloves? • What to do in case you are involved in a fire or chemical emergency?

CHP • Cal/OSHA requires that all laboratories have a written Chemical Hygiene Plan • It is on the yellow flip chart- that we all are required to sign as soon as we complete a safety training • It contains recommended emergency procedures, information on chemical hazards, a list of precautions to control exposure to hazards, phone numbers for emergency contacts, and other legally required information

MSDS: Material Safety Data Sheets 1. Chemical product information: chemical name, trade name, chemical formula- manufacturers information/ address etc. 2. Hazardous ingredients: TLV (Threshold limit value) and PEL (Permissible exposure limit) will give you highest airborne concentration to which a person can safely be exposed to during a normal workday 3. Physical data: Color, appearance, boiling point vapor density etc.

MSDS continued… 4. Fire and explosion data: Flash point- will tell you the temperature at which the chemical will release enough flammable vapor to ignite. This section also gives chemicals upper and lower flammability limits, special fire fighting procedures, any unusual fire or explosion hazards 5. Health hazard data: Gives acute/ chronic- health effects associated with overexposure to the chemical through inhalation, skin contact or ingestion. Includes emergency first aid procedures.

MSDS continued… 8. Special protection information: lists special protection equipment/ clothing (respirator, mask, gloves, eye protection etc. ) 7. Special precautions: discusses special precautions to be taken during handling and storage of the chemicals. Any other health and safety concerns not mentioned elsewhere on the MSDS Search for MSDS at: www. ucop. edu/riskmgt/ohp/msds. html

MSDS continued… 6. Reactivity data: ability to react with and compatibility with other chemicals, Also mentions if it has tendency to explode or release toxic fumes. 7. Spill or leak procedure: Procedure to follow in case of accidental spill/ release. Gives protective equipment needed to contain or clean-up he spill. Information on proper disposal

Chemical Inventory • Required by the office of Environmental, Health and safety (EH&S) • Gives storage information/ location of chemicals- After using chemicals please put the chemicals back in their appropriate location • Assists emergency responders Regina is working on it- so please bring any new material/ chemical to her and Katie’s attention, please do not put it in storage cabinets/ shelf by yourself (Just put the date as soon as you receive the chemical)

Chemical storage Safe storage of chemicals is a necessity in every laboratory in order to: Provide for effective management of chemicals. Lessen the risk of fire. Prevent accidental mixing of incompatible chemicals in emergencies. Minimize exposure to corrosive and toxic chemicals.

Container labels Labels on containers of purchased chemicals include: The common name of the substance. An appropriate hazard warning. Other label information may include procedures for: Proper handling. Storage. Emergency response

NFPA labels Chemical containers/ entrances to labs / chemical storage areas. Blue indicates the health hazard. Red indicates the fire hazard. Yellow indicates the reactivity hazard. White gives special information such as water or oxidizer incompatibility. Degree of the hazard is rated from 0 to 4 4 being the greatest hazard and 0 indicating no significant hazard.

Transporting Chemicals • Know the hazards associated i. e. read MSDS before getting ready to transport chemical. • Wear protective clothing/ gloves etc. • Hazardous chemicals must be attended at all times while being transported. • Use secondary containment like plastic tubs, bottle carriers and liquid tight carts having lips in all four sides. Always use sturdy carts. • Never put non-compatible chemicals in the same secondary containers. • Do not use crowded walkways/ elevators while transporting hazardous chemicals.

Unwanted Hazardous Chemicals EH&S picks up for: • Reuse • Recycling • Disposal Contact Bill Byrne for disposing chemicals / chemical-wastes

Drain Disposal Do not dispose any hazardous material in the sinks • It interferes with the biological processes of sewage treatment • Could be released to San Francisco Bay where they are toxic to aquatic organisms or where they accumulate in Bay sediment

Drain disposal of chemicals • Create hazards of fire, explosion, or local air pollution or stench • React with other chemicals to form hazardous gases, • Corrode lab and building plumbing, • Leak out of old pipes as liquid to pollute campus grounds, • Escape from sewer pipes as air pollutants • Expose plumbers to contact or inhalation hazards.

Drain Disposal continued… • In very small quantities certain water soluble chemicals and organics could be disposed into the sinks {e. g. Class A-solute only (100 gm/lab/day) and Class B-solute only (1 gm/lab/day)} However, do not do so unless you read the document and check the list of Class A and Class B chemicals http: //www. ehs. berkeley. edu/pubs/guidelines/draindispgls. html

Exceptionally Hazardous Chemicals 1. Mercury 5. Trichloroacetic acid 2. Ether 6. Perchloric acid and perchlorates 3. Nitric Acid and Nitrogen oxides 4. Phenol 7. Acrylamide

Personal Safety • Know the nearest emergency eye-wash and shower • Never Pipette by mouth • Do not use any equipment unless you are familiar with its working • If you are leaving a lab unattended turn off all the burners and lock all the doors • Working alone in the lab is not recommended- have a lab-buddy • Smoking is not allowed inside buildings

Always protect your eyes Protection against splattering chemicals, airborne objects and dangerous light Safety glasses with side eye shields Splash goggles Full face shields: wear goggles underneath if you are handling strong corrosive chemical

Eyewear continued… The eyewear must meet standards put forth by ANSI (American National Standards Institute) Such eyewear has Z 87. 1 imprinted on the frame or lens

Most common Hazard to your Eyes Splashing liquid droplet while popping open an Eppendorf/ microfuge tube!!! 1. At the eye-wash station: Forcibly hold your eye open and rinse the inner part for 15 minutes 2. Obtain medical attention 3. Report to Katie, Archana or Dr. Leighton

Gloves Know the characteristics of the different glove types and understand the hazards of the chemicals involved in each procedure Always wash your hands thoroughly with soap and water: when changing into fresh gloves after working with any hazardous substances before eating or handling food/ coffee mugs after you finish all your work and are ready to leave for the day

Choosing the correct gloves NO GLOVE MATERIAL IS IMPERMEABLE TO ALL CHEMICALS!!! Consult the MSDS to choose the right gloves! Natural rubber/ latex Acetone, Alcohols, Alkalies and Caustics, Ammonium fluoride, Dimethyl sulphoxide (DMSO), Phenol, Plating solutions Neoprene Alcohols, Alkalies and caustics, Cellosolve, Degreasing solvents, Mineral acids, Oils, Plating solutions Nitrile rubber Alcohols, Ammonium fluoride, Freons Hexane, Hydrofluoric and hydrochloric acid Perchloroethylene Phosphoric acid, Potassium and sodium hydroxide

Gloves continued… • Always wear gloves for handling media or liquids containing Se- compounds • Always remove gloves before Answering telephone Walking in the hallway (unless you are carrying something that requires gloves). Touching anything like keyboards, door-knobs etc.

Allergies due to Latex Gloves Not uncommon- could appear after repeated use Rash that appears on hands and stops at wrists Increased risk to people with a history of asthama, allergic rhinitis, eczema Try nitrile gloves or wear cotton glove liners Dry hands thoroughly after washing and apply topical medicated ointment/ hand lotion

Protective clothing/ Lab coats Consult the MSDS for specific material (choice of disposable lab-coats): Lab coats as essential • to avoid burning hazard: while working with an open flame, heating block, autoclave or while pouring media • for protection against spills: of toxic/corrosive chemical and biological cultures • to avoid entanglement of clothes in machinery: especially if you are wearing loose clothing Do not wear them outside the lab- in public areas!!!

Shoes should be worn at all times Not mandatory by recommended: Sandals, open-toed shoes, and shoes with woven uppers, increase the risk of exposure due to the danger of spillage of corrosive or irritating chemicals.

Respiratory protection/ Safety Masks • While working with hazardous chemicals always choose the fume hood • To avoid irritating (non-toxic) dust/powder from getting into your respiratory system choose the blue dust masks • Wear the mask first and then the gloves. • Always remove the gloves first, then wash your hand then remove the mask • Consult the MSDS to check if you require special kind of masks with specific filters etc.

General lab safety • Keep aisles clear in the lab • Maintain unobstructed access to all exits, emergency showers, eyewashes, fire extinguishers and electrical panels • Bottle, burners, equipment, heating blocks etc should be kept away from edges of counters • Never leave gas burners and hot plates unattended

Summary: Practicing safe science • Know your protocol, execute procedures meticulously • Know about all the chemicals involved- read MSDS • Use smaller quantities or choose safer chemicals if possible • Avoid splash hazards- do not flick open eppendorfs • Wear safety clothing, gloves, glasses • Use chemical hood to protect from fumes or for handling large quantities of hazardous chemicals • Be alert and maintain concentration

Using Chemical / Fume Hood Chemical Hood

Using laboratory chemical hoods • Plan your experiment: assess the risks of your experiment and place everything you need in the hood before starting. • Lower the sash. Always make sure the sash is pulled down to the marked level. The best protection is provided when the sash is brought to the lowest level for convenient operation. • Watch your activity. Use slow and deliberate motions. Place supplies so that they do not obstruct the airflow at the airfoil sill or the exhaust slots at the back of the hood. Do not work within four inches of the airfoil sill.

Performance of Chemical Hoods The performance of a chemical hood can get compromised due to: • Room air drafts/ open doors/ open windows • rapid hand movements into and out of the hood, • obstructions at the airfoil sill can affect. • someone closing a door / walking by can create drafts that cause vapor to escape from the hood. Suggestion: When planning to do a hazardous experiment, you should ask your colleagues to reduce activities such as walking past the hood or opening and closing the lab doors while you are working.

Emergencies • Always be prepared! • Emergency response: Know the locations of Eyewash fountains and emergency showers. Fire Extinguishers Emergency telephone numbers. Emergency exits and evacuation routes. Spill kits.

Fires Fire is the most potentially devastating emergency in the modern biology laboratory. It is imperative that you know how to prevent fires and be prepared to respond should a fire occur.

Preventing fires Use of flammable solvents is a primary cause of lab fires • Use the smallest quantities of flammable solvents practicable. • Store stock quantities in flammables storage cabinets • Separate flammable solvents from sources of ignition: never use a Bunsen burner in any area where flammable solvents are handled.

Fighting small fires • Always pull the fire alarm first • Turn of any electrical equipments/ burners in the area and stay on the exit side of the room (near a door) • Most of the extinguishers are labeled BC or ABC- direct the extinguisher at the base of the fire • If you are unable to put of the fire try to confine it • If you are able to put of the fire stay around to talk to UCPD and to make report

Types of fire extinguishers • Class A Extinguishers will put out fires in ordinary combustibles, such as wood and paper. • Class B Extinguishers should be used on fires involving flammable liquids, such as grease, gasoline, oil, etc. • Class C Extinguishers are suitable for use on electrically energized fires. • Class D Extinguishers are designed for use on flammable metals and are often specific for the type of metal in question.

Clothing Fire Your co-worker spilled ethanol and his/ her clothing caught fire: 1. Put out the fire: Use Blanket or make him/her roll on the floor to smother fire 2. Chill the burns with ice water 3. Seek medical attention (Tang or Alta Bates) and report the incident to your supervisor

Emergency procedures: Major lab fire 1. Alert people to evacuate the area 2. Activate nearest fire alarm or call for emergency response. 3. Close lab doors to confine fire. 4. Have a person who is knowledgeable about the lab and the incident ready to assist emergency personnel.

Emergency procedures: Evacuation after a Fire Alarm • Leave immediately if you hear fire alarm. • Always close your door, electrical equipments, burners • Do not use elevators • Go to the designated meeting areas • Do not re-enter unless the fire department declares that it is safe to do so

Emergency procedures: Earthquake • Duck under nearest sturdy object until shaking stops • If you are in a doorway- brace yourself against the frame • Avoid tall book cases/ shelves • Exit when shaking stops- always use staircases • Do not operate electrical appliances/ burners- due to possible gas leaks

Emergency procedures: Bomb Threat • Report any Bomb Threat ASAP to Carol or UCPD • Be calm and evacuate immediately as directed by emergency personnel- Take your belongings with yourself • Check work area for unfamiliar items as you leave

Hazardous Chemical spills Do not attempt to clean the spill • If you lack the knowledge to do it safely • You do not have the appropriate material required for clean-up • The spill is larger than 1 liter • You feel physical symptoms: eye irritation, breathing difficulty, dizziness, nausea etc.

Emergency procedures: Hazardous Chemical spills • Isolate the spill area • If the spill is very large- notify the people in labs directly below your lab • Refer to the yellow Flip chart • Call EH&S or 9 -911 after Hours/ holidays • Be ready with the MSDS information for that material

Emergency procedures: Medical • • Remain calm. Initiate lifesaving measures if required. Call for emergency response. Do not move the person unless there is danger of further harm. • Keep the injured person warm.

Emergency Response: Eye Exposure • First aid: Flush at the eye-wash station for minimum 5 to 15 minutes maximum by forcing the eye(s) open • Minor injuries- Tang Center • Major Injuries- Alta Bates: Have someone who knows about the incident accompany the victim Report to Dr. Leighton / Katie/ Carol

Emergency Response: Skin Exposure • First Aid: Wash exposed area (use Shower if required) for minimum 5 to 15 minutes maximum. Get help if possible. • Remove contaminated clothing to protect skin from further exposure • Wrap the person in a dry blanket after the shower • Tang center / Alta bates • Report the incident to Dr. Leighton/ Katie/ Carol

Emergency Response: Respiratory Exposure • Bring the victim to fresh air • Life-threatining injuries to be reported to 911 or 9911 • Non-life-threatening to Tang Center/ Alta bates • Inform the medical responder about the nature of the air-borne hazards so they may take precautions during resuscition procedures

Ethidium Bromide • EB is a potent mutagen: absorbed through skin, nose, eyes and mouth (Read MSDS) • Pure EB should only be handled in a fume hood: with the user wearing protective equipment that includes a lab coat, closed-toe shoes, chemically resistant gloves, and chemical safety goggles (not just safety glasses) • Nitrile is an effective barrier to short-term exposure to EB. Use 100% nitrile gloves! • Wash hands after removing their gloves, even if the gloves are not punctured

Substitutes for Ethidium Bromide? New fluorescent dyes have been developed that manufacturers, such as Molecular Probes Inc. and FMC Bioproducts, claim are less toxic and can detect nucleic acid components at lower concentrations than EB.

Exposure to Ethidium Bromide • Eyes: immediately go to the emergency eyewash flush eyes by forrcing them open for at least 15 • Skin contact: immediately wash the affected area with soap and cold or cool water for 15 minues. • inhalation of EB dust: move him to an area where he can breathe fresh air. • In all three cases the affected person should immediately seek a medical evaluation from Tang Center or from Alta Bates Hospital if the exposure occurs while Tang Center is closed

Laboratory Injuries and Physical hazards to Research workers

Avoiding Physical injuries 1. Good laboratory practice and common sense: Staying alert- take a break and relax if you are tired. Thinking about where the hazards are. Keep floors dry and uncluttered to prevent slips, trips, and falls Whenever possible, open flames should be replaced by electrical heating

Minor Cuts and Injuries. • First aid: Vigorously wash with soap and water for several minutes • Apply first aid antibiotic/ ointment • Obtain medical attention- ever if there is no exposure to hazardous substance • Report to Dr Leighton/ Katie/ Carol

Muscle and Bone Injuries • The second most common serious injuries: strains and fractures due to falls and improper lifting of heavy objects

Specific Hazards • Autoclave Hazards • Centrifugation Hazards • High Voltage equipment hazards

Autoclave Hazards Physical hazard due to steam, heat and pressure • Make sure seals on containers of liquids are loose so vapor expanding during heating will not cause an explosion • Never autoclave any flammable or volatile liquids because they could explode ( never autoclave anything that has bleach on it) • Never autoclave anything that says nonautoclavable or if you are not sure • For specific instructions: Contact Katie

Using autoclaves • Use loading racks: Never place any item directly on the bottom or floor of the autoclave • Starting the sterilization cycle: close the door of the autoclave fully choose correct cycle before starting the sterilization • Unloading the autoclave: ü wear heat-resistant gloves ü wait for the “complete” sign ü First, slightly crack open the door -wait a full five for autoclave loads with just glassware and ten minutes before unloading liquids. ü let the glassware stay outside for 15 minutes before handling the individual pieces.

Centrifugation Hazards • Personal injury is infrequent and usually associated with improper lifting techniques to move heavy rotors • 90 % of the problems occur due to user errors • Carelessness can result in sample loss or damaged equipment • It require careful use, maintenance and careful bookkeeping

High Voltage Equipment Electrophoresis equipment • Read all the instructions carefully- right next to the electrophoresis equipment or at EH&S website: http: //www. ehs. berkeley. edu/pubs/helpsheets/04 electro. html

Electrophoresis Equipment General safety with electrophoresis equipments: • Switch power off before making any connection • Handle power leads one at a time and with one hand only- make sure your hands are dry • Beware of loose connections- they lead to sparks • Keep equipment away from sink or wet area • If you feel a tingle when you touch the unit- stop using it or wait until it is repaired

Working with microbes • In our labs in Mulford hall (303/307 and 330) we work strictly with non- pathogens • Any known or suspected pathogens, their DNA’s, amplicons etc. should be handled in LL 45 in Koshland hall. This is a designated area with limited access and all people using this area should read “Leighton Laboratory Bio-safety Manual” and discuss specific issues with Dr. Leighton or Archana

Pipetting technique • Never pipette by mouth!!! • Discharge pipettes against the flask or tube wall to avoid splashes. • Use disposable pipettes only for microbial cultures and procedure (use glass pipettes for organic chemicals and corrosive agents) • Pipettes used for any microbial cultures should be treated with 10 % bleach solution before discarding them as biohazard waste • Leave them in brown box separately for trash pick-up (Ask Bill if you do not know the location of this box) • Dispose glass pipettes and broken glass in “Broken glass containers”

BIOHAZARDS • Stay away from designated refrigerators, freezers and room marked with this sign unless you are authorized to work in such areas (you are working with known/ suspected pathogens!)

Non- medical sharp waste • Sharps such as blades, needles, scalpels should be handled as “Biohazard sharp wastes” • Placed in puncture-resistant containers labeled “ Sharps waste” or “Biohazard” • Do not over fill these containers • Tape them close and call EH&S for a pick-up!

Autoclaving Biotechnology wastes (non pathogenic plates only) • Dispose in white designated bins lined with white autoclavable bags labeled non-medical wastes • Fill only to 2/3 rd for proper sterilization • Double-bag and make 2 small slits in the inner bag - close both bags without making them air-tight • Autoclave for 35 minutes on liquids after 5: 00 p. m on a weekday ( weekend: anytime) • Dispose in regular trash in the dumpsters on the North side of the building

Drain Disposal of Non. Pathogenic wastes • Liquid cultures and stocks of non-pathogenic bacteria could be treated with 10% bleach for 30 minutes before disposing in the drain. • Never dispose cultures or stocks of pathogenicbacteria (even after treating with bleach) • Never put bleach-containing solution in autoclave this could cause explosion!!!

Disposing Biohazard Wastes • Sharps: are contained in leakproof, rigid, puncture -resistant, durable plastic containers. These containers are red in color and equipped with a tight-fitting lid. EH&S to be notified for pick-up. • Free flowing liquid waste: is treated with chlorine bleach (10% final concentration) and contained in leakproof, rigid durable containers for EH&S pick -up • Solid biohazardous waste is collected in plastic waste bags (red or white) to be placed into the autoclave. White is disposed with trash and red should be disposed in designated bins for EH&S pick-up.

Important Contacts • • Dr. Leighton Carol Wehr Katie Wheeler Archana Sudame EH&S UCPD (emergency) UCPD (non-emergency)

References • • http: //www. ehs. berkeley. edu/ http: //www. practicingsafescience. org (Yale) http: //rehs. rutgers. edu/ (Rutgers) http: //www. pp. okstate. edu/ehs/index. htm (Oklahoma State)