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LABORATORY SAFETY Lisa Bognar Phillips

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1 LABORATORY SAFETY Lisa Bognar Phillips
Risk Management & Safety Including Emergency Response, Personal Protective Equipment and Hazardous Waste Management Training

2 Chemical Hazards There are two general hazard classes that must be evaluated when assessing the safety of a chemical: Physical Hazards Health Hazards

3 Chemical Hazards: Physical Hazards
Compressed Gas - cylinders of compressed gas are under intense pressure. Corrosivity – strong acids and bases can cause visible destruction of metals at the site of contact. Flammability - both solid and liquid chemicals can serve as fuel sources to support a fire. Reactivity - many substances will react violently if allowed contact with air, water or other chemicals. Friction, shock, light or heat can be enough to trigger some reactions. Oxidizability – compounds that release oxygen as they decompose. React with flammables

4 Chemical Hazards: Health Hazards
Irritants - cause reversible inflammation of skin, eyes or nasal passages. Sensitizers - cause allergic reaction after repeated exposures. Individual sensitivities vary. Carcinogens - alter DNA or cause cells with altered DNA to multiply. Systemic Toxins - typically target a specific organ or system (liver, kidney, blood, nervous system, reproductive system, etc.). Toxins – based on dose. Anything can be poisonous. Corrosives – cause burns to the skin, eyes, mucous membranes and respiratory tract. HF is not like typical mineral acids. It is extremely toxic and penetrates to the bone. Calcium gluconate is the only antidote that should be used on an HF burn.

5 Routes of Entry Inhalation: The major route of entry when working with solvents. Great rapidity of absorption

6 Routes of Entry Absorption: May produce systemic poisoning. Condition of skin determines rate of entry. Examples: organic lead, solvents, (xylene, methylene chloride) organo phosphate pesticides, cyanides

7 Ingestion: Usually accidental or due to poor hygiene practices
Routes of Entry Ingestion: Usually accidental or due to poor hygiene practices

8 Chemical Right to Know Product Information Fire and Explosion
.MSDS Includes the following: Product Information Fire and Explosion Toxicology Health Effects PPE Storage Leaks and Spills Waste Disposal First Aid )

9 Product Information

10 Finding an MSDS Chemical manufacturers and distributors CANNOT refuse a request for an MSDS. There are excellent on-line MSDS databases:

11 University of Notre Dame Chemical Hygiene Plan (CHP)
Contains information and procedures that laboratory personnel can use to protect themselves from the chemicals they work with First place to look for answers to questions you might have regarding chemical or laboratory issues Located on Risk Management and Safety website Topics include: - Emergency Response - Personal Protective Eq. - Chemical compatibility -Waste Management - Standard Operating Procedures (SOPs)

12 Personal Protective Equipment
Gloves are required anytime you are handling chemicals, biologicals and radioactive materials Eye protection Open toed shoes do not protect your feet and ARE NOT allowed in the laboratory. (NO FLIP FLOPS, SANDALS, etc)

13 Wear Eye Protection When Using:
Glassware under reduced pressure Cryogenic materials Glassware under elevated pressure Explosives Caustics, Irritants or Corrosives Biohazards Radioactive Materials UV Light Toxic Chemicals Carcinogens Flammable Materials Lasers

14 Gloves Be sure you know which type of glove is appropriate for the chemical you are using Example of wrong glove having been used

15 Gloves Remove soiled gloves. If gloves become soiled or contaminated
during experiment, stop, and remove carefully. Wash your hands and put on new pair of gloves. If you double-glove, remove soiled glove, replace with new one, and continue your work.

16 Safety Equipment: Chemical Fume Hood
Airflow into the hood prevents chemicals inside from migrating out into your breathing zone. If air velocity into the hood is impeded or slowed, the hoods ability to capture chemicals is compromised. Factors that affect airflow: Sash Height Drafts Bulky objects inside

17 Gas Cylinder Safety Storage and Handling
Gas cylinders should not be stored in exits or egress routes or blocking any safety equipment Gas cylinders (excluding lecture bottles) should be stored in an upright position and with safety caps in place unless in use. Use only the appropriate regulator for the gas Gas cylinders must be secured with a chain or appropriate belt above the midpoint but below the shoulder of the cylinder

18 What’s Wrong in this Picture?
Cylinder Safety What’s Wrong in this Picture? Cylinder must be secured by chain or other steadying device


20 Electrical Safety Eliminate frayed or worn wiring
Never stretch wires across floor or other equipment. Staff should know location of circuit breakers

21 Electrical Safety Match size of extension cord to appliance power cord to prevent cord overheating. Extension cords are not intended for "permanent" installations -- appliances shall be connected to permanently wired receptacles.

22 Examples of Poor Storage
Never store a chemical with a missing or obscured label )

23 Container Labels All chemicals (including solutions and chemicals transferred from their original containers) should be labeled with their common names, concentrations and hazards. Additional information such as: date received, date opened, date of expiration should also be recorded. Label may also include: Proper handling, Storage, Emergency response

24 Refrigerators and Freezers
Ordinary household refrigerators and freezers constitute a hazard when used for storage of flammable or unstable chemicals. These units produce conditions that can lead to explosions. Domestic refrigerators should not be used for flammable chemical storage. "Lab-safe" refrigerators and freezers (designed for storage of flammable liquids) must be used for flammable chemicals.

25 Definition Biosafety The application of combinations of laboratory practice and procedure, laboratory facilities, and safety equipment when working with potentially infectious microorganisms. We use these practices to protect the workers and those around, the product and the environment. Four biosafety levels (BSLs) are described which consist of combinations of laboratory practices and techniques, safety equipment, and laboratory facilities. Each combination is specifically appropriate for the operations performed, the documented or suspected routes of transmission of the infectious agents, and the laboratory function or activity. Descriptions of required biosafety practices and procedures are presented in both the CDC/NIH manual, “Biosafety in Microbiological and Biomedical Laboratories,” and the WHO manual “Laboratory Biosafety Manual.” Laboratories should have copies of both manuals available for workers.

26 Laboratory Requirements
Knowledgeable supervisor Knowledgeable personnel Aware of potential hazards Proficient in lab practices & techniques Availability of a lab specific biosafety manual Contact information Copies of IBC,IRB,RCC, and/or IACUC protocols Standard Operating Procedures For unique procedures Laboratory inspection checklist Emergency response plan Special procedures where needed Relevant reference materials The most important element of containment is strict adherence to standard microbiological practices and techniques. Persons working with infectious agents or potentially infected materials must be aware of potential hazards, and must be trained and proficient in the practices and techniques required to handle such material safely. The director or person in charge of the laboratory is responsible for providing or arranging the appropriate training of personnel. Each laboratory should develop or adopt a biosafety or operations manual that identifies the hazards that will or may be encountered, and that specifies practices and procedures designed to minimize or eliminate exposures to these hazards. Personnel should be advised of special hazards and should be required to read and follow the required practices and procedures. A scientist trained and knowledgeable in appropriate laboratory techniques, safet procedures, and hazards associated with handling infectious agents must be responsible for the conduct of work with any infectious agents or material. This individual should consult with biosafety or other health and safety professionals with regard to risk assessment.

27 Classification of Infectious Agents
Found in the NIH Guidelines. Classified into risk groups on the basis of risk to the individual and to the community. Currently, 4 risk group levels have been designated. the least risk (RG-1) to the most risk (RG-4).

28 RG-1 Agents RG-2 Agents Not associated with disease in healthy adult humans. Generally only require a laboratory with minimal containment Biosafety Level 1 laboratory “Declaration of Dangerous Goods” is not required for shipment. Bacillus subtilis, Saccharomyces cerevisiae, Staphylococcus epidermidis, Escherichia coli K-12 Associated with human diseases which are rarely serious. Preventative and therapeutic interventions are available. Generally require a laboratory with moderate containment. Biosafety Level 2 laboratory “Declaration of Dangerous Goods” required for shipment. Measles virus, Salmonella spp., Toxoplasma spp., Hepatitis B virus, leshmainia, plasmodium falciparum

29 Biosafety Levels of Containment
Combination of laboratory practices, safety equipment, and laboratory design to achieve levels of physical containment. Currently, 4 biosafety levels have been designated. The least restrictive (BSL-1) to the most restrictive (BSL-4). The recommended biosafety level(s) for the organisms represent those conditions under which the agent ordinarily can be safely handled. The laboratory director is specifically and primarily responsible for assessing the risks and appropriately applying the recommended biosafety levels. When specific information is available to suggest that virulence, pathogenicity, antibiotic resistance patterns, vaccine and treatment availability, or other factors are significantly altered, more (or less) stringent practices may be specified.

30 Containment: BSL-1 BSL-2
RG-1 Agents Not known to cause disease in healthy adult humans Practices Standard microbiological practices Safety equipment None required Facilities Open bench top RG-2 Agents Associated with mild to moderate disease in humans Practices BSL-1 plus limited access. Safety equipment Biological Safety Cabinet and personal protective equipment as needed. Facilities BSL-1 plus the availability of a mechanism for decontamination.

31 BSL-1 Standard Microbiological Practices
Restrict or limit access when working Prohibit eating, drinking and smoking Minimize splashes and aerosols Decontaminate work surfaces daily Decontaminate infectious waste Maintain an insect & rodent control program Prohibit mouth pipetting/ use mechanical pipetters Needles & sharps precautions Use sharps containers DON’T break, bend, re-sheath or reuse syringes or needles Use alternatives to needles when available DON’T place needles or sharps in regular trash containers. Access to the laboratory is limited or restricted at the discretion of the laboratory director when experiments or work with cultures and specimens are in progress. A biohazard sign can be posted at the entrance to the laboratory whenever infectious agents are present. The sign may include the name of the agent(s) in use and the name and phone number of the investigator. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human use are not permitted in the work areas. Persons who wear contact lenses in laboratories should also wear goggles or a face shield. Food is stored outside the work area in cabinets or refrigerators designated and used for this purpose only.

32 BSL-1 Standard Microbiological Practices
Wash hands Persons wash their hands after they handle viable materials, after removing gloves, and before leaving the laboratory.

33 BSL-1 Safety Equipment (Primary and Secondary Barriers)
Personal protective equipment Eye wear Gloves Lab coat Countertops and floors easily cleaned and decontaminated Laboratory Requirements: Lab does not need to be separated from other lab sections and can be of normal construction. No specific ventilation req. Protective eyewear should be worn for conduct of procedures in which splashes of microorganisms or other hazardous materials is anticipated.

34 Training Requirements BSL-1 BSL-2
Supervisor Trained in microbiological or related science practices. Lab personnel Specific training in lab procedures. Supervisor Is a competent scientist Restricts access Lab Personnel Aware of potential hazards Proficient in practices and techniques

35 BSL-2 Standard Microbiological Practices
Follow all practices and personal equipment used for BSL-1 containment.

36 BSL-2 Safety Equipment (Primary and Secondary Barriers)
Class II Biological Safety Cabinet recommended to protect product, personnel, and the environment. Requirements: Laboratories have lockable doors and separated from public areas Sink for hand washing Work surfaces easily cleaned Bench tops are impervious to water Sturdy furniture Adequate illumination Air flows into lab without re-circulation to non-lab areas Windows fitted with fly screens Protective eyewear should be worn for conduct of procedures in which splashes of microorganisms or other hazardous materials is anticipated.

37 Biological Safety Cabinets as Primary Barrier- Overview
Protection of Product Personnel Environment Class II Biological Safety Cabinet Equipment is laid out to not restrict airflow in the cabinet “High efficiency particulate air” (HEPA) filter Traps particulates 0.01 microns and larger including bacteria and viruses chemicals, fumes, and vapors pass through Properly maintained Class I and II BSCs, when used in conjunction with good microbiological techniques, provide an effective containment system for safe manipulation of moderate and high-risk microorganisms (Biosafety Level 2 and 3 agents). Both Class I and II BSCs have inward face velocities ( linear feet per minute) that provide comparable levels of containment to protect laboratory workers and the immediate environment from infectious aerosols generated within the cabinet. Class II BSCs also protect the research material itself through high-efficiency particulate air filtration (HEPA filtration) of the air flow down across the work surface (vertical laminar flow). Class III cabinets offer the maximum protection to laboratory personnel, the community, and the environment because all hazardous materials are contained in a totally enclosed, ventilated cabinet.

38 Biological Safety Cabinet - Operation
1. Load with needed supplies 2. Turn on and allow to run for minutes 3. Check inward airflow with a piece of tissue 4. Enter straight into cabinet and perform work in a slow, methodical manner 5. At end of work package or decontaminate items to be taken out of cabinet 6. Decontaminate interior of cabinet 7. Allow cabinet to run for minutes Shut off Annual certification is required.

39 What happens when an open flame comes in contact with ethanol in a Biosafety Cabinet

40 BSL-2 Facility Design (Secondary Barrier)
Signage on the door with restricted access when work in progress or agents are present. Autoclave available Eyewash station available

41 Decontamination Definitions
Sterilization physical or chemical procedure used to destroy all microbial life, including large numbers of highly resistant bacterial spores. Disinfection physical or chemical procedure used to eliminate all recognized pathogenic microorganisms but not all microbial forms (bacterial spores) on inanimate objects. Antiseptic germicide that is used on skin or living tissue for the purpose of inhibiting or destroying microorganisms.

42 Decontamination: Conclusion
Disinfectants do not replace adherence to good standard microbiological practices and proper hygiene!

43 Biohazardous Waste stock cultures
materials containing or contaminated with blood blood and body fluidsSerum, Plasma Other blood components Semen Vaginal secretions Cerebrospinal fluid Pleural fluid Peritoneal fluid Pericardial fluid Amniotic fluid Any other body fluid visibly contaminated with blood sharps Pipette/pipette tips, Hypodermic needles, Scalpels, Breakable containers, Glass products (i.e., slides or cover skips) animal waste other contaminated lab material

44 Biohazardous Waste Containers
Biohazardous waste containers shall be clearly marked with the universal biohazard symbol. Puncture-proof, leak-proof, sealable receptacles avoid over-filling

45 Orange Biohazard Bags Orange Biohazard Bags contain previously contaminated waste that has been decontaminated (autoclaved) by lab personnel. May be taken to the dumpster if brown bagged

46 Emergency Response Fires/accidents 911 Spills 631-5037
(From a cell phone ) Spills (Risk Management and Safety during working hours) After Hours Spills 911 State the type of emergency and the exact location of emergency. If a chemical is involved spell the chemical name to the dispatcher.

47 Alert persons in area of fire Close doors to confine fire
Fire Safety Alert persons in area of fire Close doors to confine fire Activate fire alarm Evacuate through nearest exit Do not use elevators Move away from the building.

48 How to Use a Fire Extinguisher
pull pin aim at the base of the fire squeeze handle sweep from side to side PASS

49 Fire Extinguisher Training
Learn how to operate a portable fire extinguisher by participating in fire extinguisher training program sponsored by Notre Dame Fire Department. Contact them at if you would like to take the class. US&A (v 2/07)

50 Biological/Chemical Exposures
Needle Sticks: Ocular Exposures: Dermal Exposures: Wash thoroughly with soap and water Rinse at eyewash at least 15 minutes Rinse under drench shower at least 15 minutes Identify source (consent?) Hold eye open to ensure effective wash Remove all contaminated clothing Seek Medical Treatment Your supervisor must fill out an accident/exposure form

51 First Aid - Chemical Burns (Eyes)
Forcibly open eyelids to ensure effective washing behind eyelid Wash from nose out to ear to avoid washing chemicals back into eye or into an unaffected eye Flood eyes and eyelids with water/eye solution for minimum of 15 minutes

52 First Aid - Chemical Burns (Eyes)
Make sure you know the location of the nearest eyewash station.

53 Eye Wash Maintenance Eyewashes are periodically checked by Facility Services, but are not flushed. Lab personnel must test eyewashes monthly. Designate someone to test eye washes monthly. Let run for 1 minute

54 Spill Response Be prepared for emergencies

55 Biological Spills/Decontamination
Cover spill with paper towels Soak towels with disinfectant (10% Bleach is effective) Wait 10 minutes While wearing PPE, clean up the spill using paper towels Notify Risk Management if you need assistance Wipe spill area with disinfectant Dispose of all clean-up materials as biological waste (red bag)

56 Chemical Spills Identify the chemical
Assess your ability to safely contain and clean up the spill: (spill size, chemical identification, PPE) I CAN safely clean it up I CANNOT safely clean it up Notify coworkers and secure the area Notify coworkers and vacate/secure the area Use spill kit to contain and clean up the spill (MSDS helps) Call During working hours After hours 911 On cell call Call Risk Management to confirm clean up

57 Chemical Waste Disposal
Satellite Accumulation Container Compatible and in good condition Has “tight-fitting” closure and container is closed unless adding waste. Label (3 things): Must be labeled as soon as first drop of waste is put into the container. Words “waste or “hazardous waste” Description or Name of chemical(s) Secondary Containment Must segregate incompatibles One container per waste stream


59 Hazardous Waste Disposal Rules
Do not use chemical symbols or abbreviations when labeling your containers US&A (v 2/07)

60 Abbreviations and formulas NOT acceptable
341 Chemistry Methanol Acetone 4 Liters Generator Signs Faculty name printed RM&S use ONLY Abbreviations and formulas NOT acceptable Person who generates waste signs form

61 All other waste… Gloves, paper, etc:
disposed of in regular (clear) trash bags and bags are to be removed by custodians daily. Broken Glass: if clean, should be disposed of in a puncture proof box marked “Broken Glass”.

62 Laboratory Security There are extremists who vandalize workplaces where they suspect animal research is being conducted and others who identify sources for chemicals/ biologicals to be used in terrorism or illegal drug production Protect your lab and your research: NEVER prop open any card or key access corridor doors. Don’t let people ‘piggy-back’ to gain access to restricted areas. Keep your lab doors closed and locked when unattended. Question strangers, and report suspicious activity to campus security.

63 Emergency Phone Numbers
Security or from cell Risk Management & Safety ND Fire

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