Orientation to the Laboratory Environment

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Required for all new laboratorians or lab staff prior to beginning work in the lab environment

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Goals

• This training satisfies these initial training requirements
• Hazard Communication
• General Chemical Safety
• General Biosafety
• Fire and Life Safety
• Lockout/Tagout Awareness

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EHS Staff

• Dr. Kristin Long-Witter, Director
• Cassandra Freeman, Administrative Specialist
• Kathleen Ingram
• Chemical & Radiological Safety Specialist
• Thomas Verrault
• Fire & Life Safety and Emergency Management Specialist
• Dr. Zuzana Drobna
• Biological Safety Specialist

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EHS Mission Statement

• Facilitate University’s mission of research, teaching, and service by ensuring health, safety, and regulatory compliance
• Qualified, knowledgeable team members are recognized as trusted subject matter experts and are committed to providing excellent service and leadership to campus partners
• Enables and empowers NCCU to adopt a culture of safety in which innovation, discovery and positive change are fully supported by safety professionals 

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Regulatory Requirements

• EHS is responsible for ensuring compliance with many different regulations
• Environmental Protection Agency (EPA)
• NC Department of Environment and Natural Resources (DENR)
• NC Department of Health and Human Services Radiation Protection Section
• Office of State Human Resources
• NC Fire Prevention Codes
• National Fire Protection Act (NFPA)
• Occupational Safety and Health Administration (OSHA)
• General Duty Clause
• Hazard Communication standard (29 CFR 1910.1200)
• Personal Protective Equipment (PPE) standard (29 CFR 1910.132)
• Bloodborne Pathogens standard (29 CFR 1910.1030)
• Eye and Face Protection standard (29 CFR 1910.133)
• Occupational Exposure to Hazardous Chemicals in Laboratories standard (29 CFR 1910.1450)
• Respiratory Protection standard (29 CFR 1910.134)
• Hand Protection standard (29 CFR 1910.138)
• Control of Hazardous Energy standard (29 CFR 1910.147)

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Responsibilities

• EHS has administrative responsibility for the laboratory safety program
• Everyone involved in laboratory operations - from the highest administrative level to the individual laboratorians - has a responsibility to themselves and to colleagues to plan and execute laboratory operations in a safe manner.
• Specific responsibilities which will foster research and ensure safety and compliance are assigned to each group.
• A positive safety culture is built on empathy and compassion and strives to encourage high quality, safe research. A positive safety culture does not blame or reprimand others, rather laboratorians recognize that administrators and faculty place their well-being above all.

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�EHS

• Provide and document training for laboratory personnel
• Inspect laboratories at least annually for safety and health hazards and for compliance with state and federal regulations
• Investigate potential safety and health hazards identified by laboratory employees
• Monitor personnel as needed for chemical, biological, physical, and radioactive hazards
• Advise laboratory personnel on proper disposal of waste chemicals, biologicals and other hazardous materials
• Consult with faculty, staff, students, and University Safety Committee on safety matters

Principal Investigator/Lab Manager

• PI may delegate many duties to Lab Manager
• PI retains all responsibility for ensuring compliance and safety
• Prepare a Laboratory-Specific Safety Plan
• Ensure that laboratory personnel meet all laboratory safety training requirements
• Ensure that proper safety supplies and equipment, such as gloves, safety glasses and/or goggles, lab coats, etc. are available for all people in the laboratory.
• Notify EHS of accidents, spills, or conditions that may warrant further investigation and/or monitoring.
• Train staff on location of safety data sheets for hazardous chemicals used in the laboratory
• Keep accurate laboratory chemical inventory
• Post appropriate hazard information
• Conduct an “exit interview” with laboratory workers prior to their departure from the lab to ensure that they have properly labeled and prepared hazardous materials

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Laboratorians and Staff

• Submit a NCCU Laboratory Worker Registration Form to EHS at hire and whenever there is a change in work location or laboratory assignment.
• Familiarize yourself with Laboratory Safety Manual and your Laboratory-Specific Safety Plan
• Work with PI or designee to complete all required initial and annual training. Submit the NCCU Initial Laboratory Training Record to EHS for approval within 30 days of hire
• Follow safety guidelines when handling hazardous materials, including the proper use of personal protective equipment
• Notify PI/Lab Manager of all accidents, spills, or other safety incidents

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NCCU Laboratory Safety Manual

North Carolina Central University (NCCU) is committed to providing a safe and healthful environment for all persons including staff, students, visitors, and the surrounding community. Personnel conduct a vast array of research utilizing hazardous materials on the campus.

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This manual provides basic information about hazards that may be encountered in the laboratory and safety precautions to prevent laboratory accidents and minimize exposure to hazardous chemicals, radioactive materials, environmental issues, biological materials, and infectious agents.

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Lab-Specific Safety Plans

• Each principal investigator must prepare a Laboratory-Specific Safety Plan for which addresses the hazards and precautions specific to the laboratory. NCCU provides a template Plan for use by individual laboratories.
• The OSHA document, 29 CFR 1910.1450 Laboratory Standard, requires a written Chemical Hygiene Plan (CHP) to help protect people working in a laboratory setting
• At NCCU, each laboratory’ CHP consists of the Laboratory-Specific Safety Plan and the NCCU Chemical Safety Plan. P mandated by the Laboratory Standard are listed below along with the location.
• Each lab has designated a Chemical Hygiene Officer

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Documents Available to Lab Personnel

• NCCU Laboratory Safety Manual
• NCCU Chemical Safety Plan
• Laboratory-Specific Safety Plan (See PI)
• Access to online or printed Safety Data Sheets (SDS) for those chemicals used routinely (See PI)
• As needed
• Respiratory Protection Plan
• Radiation Safety Plan
• Exposure Control Plan

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Risk Assessment and Hazard Mitigation

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Risk Assessment

• Hazard Identification - identify hazards and risk factors that have the potential to cause harm or damage
• Analysis - evaluate the risk associated with that hazard
• Mitigation - Determine appropriate ways to eliminate the hazard or control the risk

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Hierarchy of Controls

The National Institute for Occupational Safety and Health (NIOSH) hierarchy of controls for safety measures

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Elimination

• Complete elimination of hazards would often defeat the purpose of the research being done inside of a laboratory

• Laboratorians should work to eliminate chemicals, materials, processes, and equipment that are unnecessary to specific experiments

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Substitution

• Think about the hazards you are exposed to.
• Can you reduce the amount of chemical and still achieve the desired result?
• Can you substitute one biological hazard for a lesser hazard (i.e. is there a BSL-1 agent that could substitute in early experiments for a BSL-2 agent)?
• Switch out processes, equipment, material, or other components where possible

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Engineering Controls

• Physically separate the laboratorian from the hazard

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Engineering Control – Biosafety Cabinet

• Prevent exposure of laboratory personnel and contamination of the lab from hazardous biological aerosols using three mechanisms:
• air barrier
• physical separation
• high-efficiency particulate air (HEPA) filtration.
• BSCs can also provide a clean work environment to protect cell cultures or sterile materials (product)
• There are three classes of BSCs
• Class I and II cabinets protect personnel and environment
• Class II cabinets also provide a HEPA-filtered laminar flow air to the work surface to protect product
• Appendix A of the Biosafety in Microbiological and Biomedical Laboratories has further details
• OSHA BSC fact sheet

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Engineering Control- Biosafety Cabinet

• Class II, Type A2 BSC
• (A) front opening
• (B) sash
• (C) exhaust HEPA filter
• (D) supply HEPA filter
• (E) common plenum
• (F) exhaust blower

BMBL 6^th Edition

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Engineering Control- Biosafety Cabinet

• Air Barrier
• Directional movement of room air past the laboratorian, and into the BSC via the work opening.
• Class II BSCs provide protection of product using HEPA filtered air flowing over the work area
• Disruption of the airflow in the BSC can compromise the integrity of the containment for both personnel and product
• HEPA Filtration
• Filters that can capture 99.97% of particles ≥0.3 µm diameter
• Removes virtually all particulates, including hazardous microbiological and some chemical aerosols, in the air stream passing through the filter
• Not effective in capturing chemical vapors, and are not considered protective against gases or vapor-phase solids/liquids.

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Biosafety Cabinet Best-Practices

• Never work in a BSC that does not have current certification
• Plan work in advance
• Place everything needed for the procedure in the BSC prior to beginning hazardous or sterile work
• Arrange materials so that clean and contaminated materials are segregated
• Conduct research in a ‘clean to dirty’ work pattern
• Remove any materials or equipment not necessary for the particular procedure
• Ensure sash remains at correct height while working in BSC
• Avoid placing materials on the air intake grille at the front of cabinet as this disrupts the protective air barrier
• Perform manipulations of hazardous materials at least 4 inches inside the cabinet and as far back in the work area as possible
• Decontaminate all items with appropriate disinfectant before removing from BSC if hazardous work is being performed
• Disinfect interior of cabinet with approved disinfectant after each use
• Limit access to BSC to one person at a time – excessive movement in and out can disrupt the protective air barrier
• Do not use an open flame in a BSC as it disrupts airflow and could damage the HEPA filter

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Engineering Control - Laminar flow bench/cabinet

• Provide product protection but no personnel protection
• Exhaust air passes over the working bench exits the unit towards the operator

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Engineering Controls – Fume Hood

• Designed to minimize exposure to hazardous chemical vapors by drawing harmful vapors away for filtration to remove dangerous vapors, and then either exhausted outside of the building or recirculated back into the lab. 
• The fume hood works by using a glass shield (sash) that opens or closes to contain vapors
• Keeps vapors away from the user’s face and the rest of the laboratory
• Blowers draw in air from the room, through a filter within the fume hood and towards an exhaust area
• OSHA Chemical Fume Hood QuickFact Sheet

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Working Safely in a Fume Hood

• Reduce obstructions to allow adequate air flow across the working surface with minimum turbulence.
• Keep at least 50% of the working surface clear, if possible
• Place containers and equipment toward the sides of the hood
• Elevate equipment and containers two to three inches above the working surface using perforated or slotted shelving
• Always work with chemicals at least six inches into the hood from the sash
• Check the airflow indicator prior to use to ensure the fume hood is drawing air. If an airflow monitor is not installed, test the hood airflow with a piece of tissue or chemwipe at the sash.
• Keep the sash at 18 inches or less from the working surface while working in the hood
• Keep laboratory doors and windows closed and limit movement in front of the hood. 
• If your hood is not working properly, do not use until the problem has been fixed. Place signage on the hood indicating it is out of service.
• Never place your head inside the fume hood when working with chemicals.

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Administrative Controls

• Safety rules and protocols for workers in the lab to follow
• Standard operating procedures and checklists
• Safety Plans and Manuals
• Training
• Warning signage

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Administrative Controls

• Contamination of food, drink, tobacco products, and cosmetics is a potential route for ingestion of a hazardous substance.
• University policy prohibits smoking inside or within 25 feet of any University building
• Designate non-laboratory areas, such as break rooms, lounges or conference rooms, as food storage and eating areas for laboratory personnel.
• Designated food item areas must be free from all research-related items, including PPE
• Wash food containers, dishes, and utensils only in sinks exclusively designated for food utensils.
• Do not use glassware or utensils used for laboratory operations for food or beverages.
• Do not use laboratory refrigerators, ice chests, and cold rooms for food storage. All lab cold storage should be posted with proper signage.

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Administrative Control – �Hazard Communication Signage

• Hazard Communication Sign must be present on all laboratory doors and other room doors where hazardous materials may be present
• The CDC/NIH Biosafety in the Microbiology and Biomedical Laboratory, 6th edition (BMBL) calls for a biohazard sign to be posted at the laboratory entry where biohazardous materials are present.
• NFPA 704 Fire Diamond communicates hazard of chemicals that could occur as the result of a fire, spill, or similar emergency.
• The radiation trefoil symbol needed when radioactive materials are stored or used in the laboratory.
• The laser symbol is incorporated when Class 3B or Class 4 lasers are present. Do not enter unless accompanied by lab personnel.
• Equipment with strong magnetic fields in laboratory. Do not enter unless accompanied by lab personnel, and limitations of entry are understood.

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Example of Hazard Sign

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Administrative Control - Laboratory Clothing

• The clothing you wear in the laboratory affects your safety
• Do not wear loose or torn clothing that can get caught in equipment, flames or chemicals
• Avoid clothing which exposes skin (e.g. shorts, skirts) not
• Restrain long hair to keep it clear of fire or chemicals and equipment
• Sturdy, closed-toed shoes should be worn at all times
• Do not wear perforated shoes or sandals
• High heels can create a trip and fall hazard
• In some labs cloth shoes may be prohibited to protect against chemical spills and splashes

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Administrative Controls - Equipment

• Research requires specialized equipment
• Procurement and installation may require changes to electrical, HVAC, noise/vibration dampening, structural loading and technology
• May also trigger required inspections, safety protocols/training, PPE, specialized maintenance, etc.
• Signed copy of Laboratory Equipment & Instrumentation Pre-Purchase Checklist  must be attached to all requisitions
• If the acquisition requires structural changes complete and submit the Facility Modification Request Form

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Administrative Controls - Equipment

• Lab equipment leaving the laboratory for surplus or disposal should be completely disinfected and defaced
• Attach a completed Equipment Safety Clearance Form attached after laboratory equipment is decontaminated and prior to:
• Requesting service or repair of laboratory equipment by non-laboratory personnel
• Removing any equipment from the laboratory for surplus or disposal

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Occupational Health Considerations

• Hazard mitigations are based on risks to healthy adults
• Immune compromised laboratorians may be at increased risk of illness and/or more serious side effects of illness
• NCCU encourages immunosuppressed individuals to consult with their health care provider to determine if accommodations or restrictions are necessary
• Students can contact Student Health for a risk consultation

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Conceptus Protection Program

• Available to all laboratory personnel for concerns regarding biological, chemical or radiological risks to pregnancy or fetus
• Employee must declare actual, suspected, or planned pregnancy to Supervisor and EHS to be afforded extra protection
• Exposure monitoring
• Review of lab safety
• Contact ehs@nccu.edu with any questions

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Lab Injuries and Illnesses

• It is the responsibility of the PI to ensure that employees and students receive prompt treatment of any injuries and evaluation of any known or potential lab-acquired infections or exposures
• If treatment requires more than first aid, campus police or 911 should be called to request medical care and/or transport
• Never send an injured employee to seek medical attention for an injury on their own or transport them yourself
• If the injured is an NCCU student, the incident occurred during normal business hours and emergency care is not needed, the student will be triaged by Student Health Services.
• Outside business hours, students should contact University Police for transport to the closest hospital emergency room
• Students must report all lab-associated incidents to the Office of Student Affairs
• If the injured is an NCCU employee, the incident occurred during normal business hours and emergency care is not needed, the employee should seek treatment at Duke Occupational Health Medicine
• If the incident requires emergency care, contact University Police for transport to the closest hospital emergency room

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Reporting and Injury or Illness

• Any state employee who experiences an occupational illness or injury including exposure is required to complete the Workers’ Compensation Employee Statement Form
• Supervisors must complete the Supervisor’s Accident/Incident Investigation Report Form.
• Forms must be submitted to the Workers’ Compensation Administrator within 24 hours of a workplace incident.

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Personal Protective Equipment

• Although PPE is the least effective of control measure, it should absolutely be used, in case other control measures fail
• The success of PPE depends in large part on whether or not laboratorians comply and utilize it properly
• Eye protection and protective clothing (e.g., lab coats and gloves) are the most recognizable and most used PPE in the lab.

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PPE - Protective Apparel

• Lab coats
• Primarily a protection for clothing
• Cotton and synthetic materials are satisfactory, whereas rayon and polyesters are not
• Do not significantly resist penetration by organic liquids
• NCCU utilizes a vendor to launder and return all laboratory coats – never take them home.
• Plastic or rubber aprons
• Provide better protection from corrosive/irritating liquids
• Rubber aprons offer protection when handling hot liquids
• Plastic aprons accumulate a considerable charge of static electricity, so avoid use in areas with flammable solvents or other ignitable materials
• Know the appropriate techniques for removing protective apparel if contaminated.
• Chemical spills on leather clothing or accessories (watchbands, shoes, belts, etc.) may readily absorb into the leather and hold the chemical close to the skin for long periods.
• Remove such items promptly to prevent or minimize chemical burns. If possible, decontaminate these items and if that is not possible discard as hazardous waste.

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PPE- Eye & Face Protection

• Students, faculty, staff, and visitors in laboratories must wear eye protective devices while in the laboratory space (regardless of anticipated eye hazards) with the exception of within desk areas where no hazards are present.
• The type of safety device required is determined by risk assessment based on the nature of the hazard and the frequency with which the wearer encounters it. PIs must determine the appropriate level of eye protection for particular tasks, and enforce eye protection rules.
• OSHA requires that the employer provide eye and face protection devices without cost to students, employees and visitors. Each department is responsible for funding its eye and face protection program. If prescription safety glasses are necessary, the employee and/or student are responsible for scheduling and payment for eye examinations to obtain a current prescription. Prescription safety glasses are available for State Employees at a discounted rate from Correction Enterprises.
• Contact lenses do not provide adequate eye protection for hazardous operations and must be worn in conjunction with approved safety eyewear.

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PPE- Safety Glasses

• Must be hardened-glass or plastic safety spectacles with side shields that comply with the Standard for Occupational and Educational Eye and Face Protection (Z87.1) established by the American National Standards Institute (ANSI). This standard specifies a minimum lens thickness of 3 mm, impact resistance requirements, passage of a flammability test, and lens-retaining frames.
• Do not wear photogrey (transition) lenses indoors in laboratory environments, because the percentage of light transmitted under normal light conditions is below ANSI standards.

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PPE - Goggles

• Goggles provide a tighter face seal than safety glasses, and are not for general laboratory use
• Use when there is a hazard from splashing chemicals or flying particles.
• When using glassware under reduced or elevated pressure
• Using glass apparatus in combustion or other high temperature operations
• Impact-protection goggles have perforated sides to provide ventilation and reduce fogging of the lens, but do not offer full protection against chemical splashes
• Use chemical goggles with splash-proof sides for protection from harmful chemical splash.

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PPE – Face Shields

• When you need to protect the face and/or throat from flying particles and harmful liquids
• Consider using a face shield or mask when operating a vacuum system (which may implode), or conducting a reaction with the potential for mild explosions.
• Always use a UV-blocking face shield when working with transilluminators or other devices that produce ultraviolet radiation

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PPE- Gloves

• Wear proper protective gloves for potential contact with hazardous materials, sharps, and hot or cold materials
• It is the responsibility of the PI to provide the proper gloves for each task in the lab and to enforce the lab glove policy including glove types and proper disposal
• OSHA has guidance on glove selection and chemical resistance
• Always remove gloves before contacting “clean” areas such as food area surfaces, or common equipment such as telephones, computer keyboards, and photocopiers.

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Glove Best Practices

• Consider double gloving when handling highly hazardous, toxic, or carcinogenic materials
• Before each use, inspect gloves for discoloration, punctures, and tears
• Dispose single-use gloves after you remove them – never reuse single-use disposable gloves
• You can dispose gloves in the regular trash if they are not contaminated with biohazards, chemicals or radionucleotides. For gloves contaminated with these substances, dispose in the proper waste stream. Do not dispose of contaminated gloves in a manner that could expose other personnel
• Never wear gloves outside of the laboratory –remove gloves and wash hands before exiting the lab
• Use secondary containment for items transported outside of the lab that should not be handled with bare hands
• Remove gloves even if you believe they are non-contaminated, as others do not know if you might have handled hazardous materials with your gloved hand(s)
• Do not use gloves past the expiration date

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Latex Allergy

• One of the most common chemicals in a lab
• Allergy occurs after repeated direct contact
• Sensitivity reaction may range from localized dermatitis (skin irritation) to an imme­diate, possibly life-threatening reaction
•  OSHA PPE standard 29 CFR 1910.132
• Employer must ensure that appropriate PPE is accessible at the worksite or issued to workers.
• Latex-free gloves must be available to workers
• If you know you have a latex allergy – discuss with your PI and EHS to make sure that proper protections can be put in place

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Laboratory Safety Equipment

• Eye wash & shower
• Indoors, emergency eyewash and safety showers are required within a 10-seconds travel distance and not more than 75 feet from where corrosive chemicals are used.
• Must be on the same level as the chemical area; there can be no stairs/ramps or blockages between the hazard and the eyewash and/or safety shower.
• First Aid Kit
• Class A kits in labs provide a basic range of products to deal with common injuries including minor burns, wounds, and eye injuries.

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Chemical Safety

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Lab Specific Chemical Training

• Required by OSHA Laboratory standard
• Laboratory must train you in the following before you start work
• location of the Chemical Hygiene Plan;
• permissible exposure limits (PELs) for OSHA-regulated substances, or recommended exposure levels for other hazardous chemicals where there is no applicable standard;
• signs and symptoms associated with exposure to hazardous chemicals in the laboratory;
• location of SDS forms and other reference materials;
• how the presence or release of a hazardous chemical is detected;
• physical and health hazards of chemicals in the laboratory work area; and
• Measures taken to mitigate hazard exposure including protective equipment, appropriate work practices, and emergency procedures

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Hazard Communication Standard �29 CFR 1910.1200

• Requires that employees be informed of chemical hazards that they work with or are present in their work area
• See full regulation here

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Elements of Hazard Communication

• Ensuring chemicals are labeled
• Maintaining current and accurate chemical inventories
• Maintaining current and complete Safety Data Sheets (SDS)
• Training of personnel by Supervisor on the chemicals that are used or are in the workplace

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2012 Changes to Hazard Communication

• Hazard classification
• Provides specific criteria for classification of health and physical hazards, as well as classification of mixtures
• Labels
• Manufacturers must provide labels that include a harmonized signal word, pictogram, hazard statement(s), and precautionary statement(s)
• Safety Data Sheets (SDS)
• Standardized format in 16 required sections
• Employers are required to train workers on label elements and SDS format

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Hazard Classification

• Appendix A defines health and physical hazards
• Appendix B includes additional parameters to evaluate health hazard data
• Appendix F pertains to carcinogens

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Labels

• Chemical manufacturers and importer must include following on labeling:
• Supplier information
• Harmonized product identifier
• Pictogram
• Signal word
• Hazard statement
• Precautionary statements
• Any chemical transferred from a manufacture’s labeled chemical container to another container must have the same label information as the original container

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Example of Compliant Labeling

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Distinct Hazard

• Chemical in which there is scientific evidence that a health, physical and/or environmental hazard may occur
• Health – acute or chronic health affects may occur if exposed
• Physical – a combustible liquid, compressed gas, explosive, flammable, organic peroxide, oxidizer pyrophoric, unstable (reactive) or water-reactive
• Environmental – poses risk or danger to the environment

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Hazard Statement

• Describes the nature of the hazard(s) of a chemical including, where necessary, the degree of hazard

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Pictograms

• Alerts users to chemical hazards to which they may be exposed
• 9 pictograms
• Symbol on white background framed within red border

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Signal Words

• Used to indicate the relative level of severity of hazard and alert user to a potential hazard on the label
• Danger – used for more severe hazards
• Warning – used for less severe hazards

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Precautionary Statements

• Describes recommended measure that should be used to minimize or prevent adverse effects.

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Safety Data Sheets

1. Identification
2. Hazard(s) identification
3. Composition/information on ingredients
4. First-aid measures
5. Fire-fighting measures
6. Accidental release measures
7. Handling and Storage
8. Exposure controls/personal protection
9. Physical and chemical properties
10. Stability and reactivity
11. Toxicological information
12. Ecological information
13. Disposal considerations
14. Transport information
15. Regulatory information
16. Other information

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• Formerly known as Material Safety Data Sheets (MSDS)
• Standardized formatting and information to 16 sections

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Formaldehyde standard 29 CFR 1910.1048

• Any persons who work with or around formaldehyde must complete the NCCU Formaldehyde Training before working with the chemical
• Employers must ensure that no worker is exposed to an airborne concentration of formaldehyde which exceeds 0.75 parts formaldehyde per million parts of air (0.75 ppm) as an 8-hour time weighted average (TWA).

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Cryogens and Dry Ice

• Cryogen - substance used to produce temperatures below -153°C (-243°F)
• Liquid nitrogen (LN2) which has a boiling point of -196°C (-321°F),
• Solid carbon dioxide or dry ice
• Converts directly to carbon dioxide gas at -78°C(-109°F)

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Working with Cryogens/Dry Ice

• Avoid skin contact – never handle dry ice or LN₂ with bare hands
• Use cryogenic gloves which are designed for working below-80°C
• Gloves need to be loose-fitting for easy removal if splashed
• Always use appropriate eye/face protection
• Vapors of LN₂ and dry ice can reduce oxygen below 19.5% which may cause unconsciousness and asphyxiation
• Do not use or store dry ice or LN₂ in confined areas without ventilation (i.e. cold rooms, etc.)
• Never place on tile or laminated counters as the adhesive may be destroyed
• Never store a cryogen in a sealed, airtight container above the boiling point temperature

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Reproductive Toxins

• Reproductive toxins – have deleterious effects to DNA in the egg or sperm and/or are harmful or lethal to fetus
• Teratogenic – harmful or lethal to fetus
• Mutagenic – harmful to genetic materials (chromosomes)
• Developing fetus may be adversely affected by lower doses than adults
• Examples
• Lead, arsenic, benzene and mercury containing compounds

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Air Contaminants standard 29 CFR 1910.1000

• Rules for protecting workers from airborne exposure to over 400 chemicals – several of which are commonly used in labs include: toluene, xylene, and acrylamide

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Chemical Spills

• Labs must be equipped with protective clothing and spill cleanup materials to respond to small low-hazard chemical spills
• Laboratories with mercury or mercury containing items must have special spill kit and training

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Control Spill Area & Evaluate Hazard

• Evacuate personnel from the immediate spill area
• Remove any injured personnel from spill area
• Remove contaminated clothing, flush skin with water, use eyewash and/or safety shower, etc. and seek medical attention
• Block off immediate spill area to restrict access
• Post signage, “Spill Area – Keep Out”.
• Eliminate any fire hazard if spill is flammable or combustible
• Make preliminary evaluation of hazard and risks and decide whether you should call EHS.
• If you are comfortable with clean up and can do it safely, continue with clean up

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Clean Spill Area

• Contain the spill
• Use spill pads to absorb liquid
• Clean the spill surfaces
• Replace used materials in spill kit
• Place all contaminated disposable materials in large plastic bag
• Double bag if necessary
• Submit request for hazardous waste pick-up
• Report spill to PI/Supervisor

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Emergency Response to Chemical Spills

• Do not try to clean up any chemical incident that involves any of the following:
• Respiratory hazard
• Threat of fire or explosion
• More than 100 mL of an OSHA regulated chemical carcinogen or a highly toxic chemical
• More than 1 liter of a volatile or flammable solvent
• More than 1 liter of a corrosive (acid or base) liquid
• As best as possible, try and determine the contents and potential hazards and call EHS (919-530-7125) immediately to report during work hours and University Police after hours (919-530- 6106).
• The EHS Chemical Safety Specialist or another trained EHS Specialist will immediately respond, assess the situation, remove the container or spill or leave in place and sequester the area until it can be safely removed.

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Hazardous Waste Collection & Disposal

• It is against most State and Federal regulations to dispose of chemicals or radioactive materials down the drain
• Complete a Hazardous Waste Pickup Request for removal of laboratory chemical waste. Labs should take care to not accumulate more than 10 bottles of chemical waste
• Refer to the Radiation Safety Manual for more information on radioactive waste pick up

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Remember......

• Always ask to see the SDS for a chemical BEFORE you work with it the first time
• SDS sheets must be accessible to employees at all times

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Radiation Safety

• Contact EHS Radiation Safety Officer if you will be working with radioactive materials or radiation containing equipment

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UV Spectrum

• Humans can not perceive UV directly
• The lens of the eye blocks most radiation in the range of 300-400 nm
• The cornea blocks shorter wavelengths
• American Conference of Governmental Industrial Hygienists
• Recommends exposure not exceed 01. μWatts/cm²

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UV Sources in a Lab

• Transilluminator - actinic range 200-315 nm
• Can cause severe eye and skin burns
• Crosslinker - actinic range 200-315 nm
• Never disable or override the door safety interlocks
• Biosafety cabinet / PCR or clean workstation
• Turn off the UV light before using
• Close the sash when the UV light is on
• It is safe to work near BSC when the light is on if the sash is closed
• Wear PPE including eye and skin protection
• Fully buttoned lab coat and gloves
• For open UV sources
• Full face shield marked with the ANSI Z87.1-1989 UV certification
• Anyone near open UV source should wear the same PPE as user
• If you develop skin or eye irritation or pain after working with a UV source, notify your Supervisor and seek medical attention

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Biological Effects of UV Exposure - Eye

• Photokeratitis - not felt until several hours after the exposure
• Photokeratitis is very painful and produces the sensation of having sand in your eye
• Also causes an aversion to bright light
• Tearing
• The effects typically last up to 48 hours but will disappear as the cells of the cornea are replaced
• Long-term effects
• UVA absorption by the lens can alter proteins in the lens and result in cataract formation

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Biological Effects of UV Exposure - Skin

• Excessive UV exposure in the actinic range (200-315 nm) produces symptoms that are comparable to sunburn
• Redness, swelling, pain, blistering, and peeling of the skin.
• You will recover from short-term skin damage, but chronic exposure to UV may increase your risk of skin cancer.

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Chemical Exposure Terms

• Local toxicity – chemical active at the point of contact
• Systemic toxicity – chemical is absorbed into the bloodstream and distributed throughout the body, affecting one or more organs.
• Acute health effects - last for a relatively short time and then disappear
• Chronic health effects - not reversible
• Acute exposure - short period of exposure.
• Chronic exposure – repeated exposure to chemical over time

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Dermal Chemical Exposure

• Dermal
• Most common chemical exposure route in laboratory setting
• Commonly result in localized irritation or dermatitis
• Can lead to systemic exposure
• Main portals of entry through the skin are hair follicles, sebaceous glands, sweat glands, and cuts or abrasions
• Chemicals can also enter the body when contaminated hands touch the mouth, nose, eyes, sores or cuts.

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Inhalation Chemical Exposure

• Inhalation of toxic vapors, mists, gases, or dusts can produce poisoning by absorption through the mucous membrane of the mouth, throat and lungs
• Can cause serious damage locally and may pass rapidly through the capillaries of the lungs and enter the circulatory system
• Can result from
• Dusts and particulates becoming airborne when transferred from one container to another.
• Grinding and crushing procedures
• Splashes created from spills and vigorous shaking/mixing including centrifugation

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Injection Chemical Exposure

• Accidents involving needles and syringes can result in injection of toxic materials through the skin
• Containers of toxic chemicals may break, resulting in hazard from contact with contaminated broken glass

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Ingestion Chemical Exposure

• Can occur when contaminated hands come in contact with the mouth, or with food items
• The laboratory environment can contaminate food items and utensils.
• NEVER mouth pipette

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Ocular Chemical Exposure

• Can occur via splash, or rubbing eyes with contaminated hands
• Few substances are innocuous with eye contact, and several can cause burns and loss of vision
• The eyes have many blood vessels, and rapidly absorb many chemicals
• Any ocular exposure should immediately be treated by 15 minutes of the exposed eye to clean water using the nearest emergency eyewash

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Biological Safety

Environmental Health and Safety

Biosafety in Microbiological & Biomedical Laboratories (BMBL), 6^th ed.

• The BMBL is a code of practice for biosafety
• Agent summary statements are included for agents that meet one or more of the following three criteria:
• a proven hazard to laboratory personnel working with infectious materials;
• high potential for causing LAIs even though no documented cases exist; and
• causes grave disease or presents a significant public health hazard.
• Agent summary statement describe hazards, recommended precautions, and levels of containment appropriate for handling human and zoonotic pathogens in the laboratory and in facilities that house laboratory vertebrate animals
• Risk assessments/summary statements are included in the Lab Specific Safety Plan
• Must review the hazard statement or risk assessment to learn about the risks associated with the biological agents and have this review documented.
• Other sources of pathogen risk assessment
• Pathogen Safety Data Sheet
• Lab-specific agent-specific risk assessment performed by PI

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Bloodborne Pathogens Standard 29 CFR 1910.1030

• Designed to protect workers from the health hazards of exposure to bloodborne pathogens (blood or OPIM)
• If your work involves these material you must complete BBP training at hire and annually

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BLOODBORNE PATHOGEN RISKS

• Blood
• Human blood or blood components
• Products derived from or containing human blood
• Human-derived cell lines
• Unfixed tissues or organs

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• Other potentially infectious materials (OPIM)
• semen�
• vaginal secretions�
• cerebrospinal fluid�
• synovial fluid�
• pleural fluid�
• peritoneal fluid�
• amniotic fluid�
• saliva�
• Any body fluid visibly contaminated with blood

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Disinfection and Decontamination

• Contact time is the amount of time an EPA-registered disinfecting product needs to be present on a surface in order to be effective against the microorganisms listed on its label.
• Allow disinfectant to remain WET on surfaces for the appropriate contact time
• Disinfectants are detailed in the Lab Specific Safety Plans.

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Biological Spill Procedures

• Spills involving biological materials, including recombinant or synthetic nucleic acids
• All personnel who perform clean-up must be properly trained in the hazards present.
• If personnel do not feel comfortable cleaning up any spill, evacuate the area and call EHS.

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Biological Spill Outside Primary Containment

• EVACUATE all personnel from the room and close the door if aerosols are a concern – wait 30 minutes to allow aerosols to settle before attempting to clean up the spill.
• ALERT others to avoid contamination (post spill sign from spill kit)
• REMOVE any contaminated clothing or personal protective equipment and place in a biohazard bag for decontamination and/or disposal. Wash your hands!
• PUT ON clean gloves, lab coat, and eye/face protection.
• REMOVE any solid/sharp objects using tongs, dustpan and broom or other mechanical device contained in spill kit.
• CLEAN UP the spill
• Place absorbent material (paper towels) over spill
• Gently pour/spray disinfectant from the outer rim of the spill inwards until absorbent material is completely saturated.
• Allow disinfectant to stand for appropriate contact time.
• Clean up the spill working in a circular motion from the outer rim of the spill to the center.
• Rinse area with 70% ethanol or water if bleach is used to remove any corrosive residues.
• Dispose of absorbent materials as biohazard waste.
• Wash hands with soap and water.
• REPORT incident to supervisor

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Biological Spill Inside Biosafety Cabinet

• Leave the biological safety cabinet blower on and begin cleanup immediately.
• Do not place your head in the cabinet to clean the spill, keep your face behind the view screen.
• REMOVE any solid/sharp objects using tongs, dustpan and broom or other mechanical device contained in spill kit.
• CLEAN UP the spill
• Place absorbent material (paper towels) over spill
• Gently pour/spray disinfectant from the outer rim of the spill inwards until absorbent material is completely saturated.
• If necessary, flood the work surface as well as the drain pans and catch basins below the work surface, with disinfectant; be sure the drain valve is closed before flooding the area under the work surface.
• Wipe cabinet walls, work surfaces, and inside the view screen with disinfectant.
• Lift the front exhaust grill and work surface; wipe all surfaces with disinfectant.
• Allow disinfectant to stand for appropriate contact time.
• If necessary, place a container under the drain valve and drain the disinfectant under the work surface into the container.
• Rinse area with 70% ethanol or water if bleach is used to remove any corrosive residues.
• Dispose of absorbent materials as biohazard waste.
• Wash hands with soap and water.
• REPORT incident to supervisor

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Biological Spill Procedure - Centrifuge

• For BSL2 agents, always use sealed safety-caps, sealed buckets or sealed rotors with O-rings
• Wait 5 minutes before opening the centrifuge following the end of a run with potentially hazardous biological material. If a spill is identified after the centrifuge lid is opened, carefully close the lid. Let aerosols settle for 30 minutes.
• Remove any contaminated protective clothing and place into a biohazard bag. Wash hands and any exposed skin surfaces with soap and water.
• PUT ON clean gloves, lab coat, and eye/face protection.
• CLEAN UP SPILL
• Keep rotors and buckets closed and transfer them to a biological safety cabinet.
• Carefully retrieve any broken glass from inside the centrifuge and/or rotor or bucket using forceps and discard into a sharps container. Smaller pieces of glass may be collected with forceps.
• Immerse rotor/buckets in 70% ethanol or a non-corrosive disinfectant for appropriate contact time. Allow to completely air dry.
• Intact tubes may be wiped down with disinfectant and placed into a new container.
• Wipe the inside of the centrifuge with disinfectant.
• If bleach is used, follow with 70% ethanol to remove any corrosive residues.
• Dispose of absorbent materials as biohazard waste.
• Wash hands with soap and water.
• REPORT incident to supervisor

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Biohazard Waste

• All biological waste generated during laboratory research must be deactivated by autoclaving or chemically treated before disposal
• Biohazard waste includes:
• Materials contaminated or potentially contaminated during the manipulation or clean-up of material generated during research and/or teaching activities requiring biosafety level 1 or 2 or animal biosafety level 1 or 2.
• Human liquid blood and body fluids.
• Human tissue and anatomical remains.
• Materials contaminated with human tissue or tissue cultures (primary and established)
• Animal carcasses, body parts, blood, fluids and bedding from animals infected with BSL2 agents.

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Liquid Biohazard Waste

• Liquid wastes may be autoclaved for a minimum of 30 minutes at 121°C and 15psi
• Chemical disinfection using an approved, agent-specific disinfectant for the manufacturer’s recommended contact time.
• Most liquid wastes can be deactivated with a 1:10 final dilution (vol/vol) of household bleach – though its important to remember that bleach is a corrosive chemical and requires an eye wash and other chemical management and training

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Solid Biohazard Waste

• Solid biological waste, e.g., pipettes, tissue culture flasks, and multi-well plates, is typically deactivated by autoclaving.
• Collect solid biological waste directly into double autoclave bags inside solid container marked with biohazard symbol
• Do not seal the bag tightly so that steam is able to enter the bag during the sterilization process
• Use a secondary container for all autoclave bags in autoclave
• Ensure the autoclave operates for at least 30 minutes at 121°C and 15psi
• Once autoclaved dispose of waste in regular trash

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Autoclave Safety

• Autoclave use can pose physical hazards (e.g. heat, steam and pressure) and biological hazards (e.g. improperly disinfected hazardous waste.
• All persons who work in a lab that has an autoclave must complete the one-time NCCU Autoclave Safety Training and quiz and received training on the specific make and model used in the laboratory before operating an autoclave.

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Biological Exposures

• Known exposure
• Needle sticks or cuts with contaminated sharps
• Bite from infected animal
• Contact of infectious materials with mucous membranes (i.e. eye, nose, mouth)
• Contact of infectious materials with broken skin
• Aerosol generating event outside of primary containment with respiratory pathogens
• Potential exposure
• Failure of personal protective equipment or primary containment with no known aerosol risk
• Direct contact with infectious materials, spills, needle stick, or cut with object not known to be contaminated
• Employee experiencing symptoms associated with pathogen that they have worked with or near
• Ingestion – eating or drinking in lab areas, handling food or cosmetics with contaminated hands or gloves

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Between 2007-2012, 60% of reported lab-associated infections had an unknown cause

ALWAYS REPORT KNOWN OR POTENTIAL EXPOSURES IMMEDIATELY!

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Biological Exposure Response

• Percutaneous exposure (needle sticks, cuts, animal bites)

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• Mucous membrane exposure
• Flush eyes for at least 15 minutes using an emergency eye wash station
• Report to Supervisor and EHS immediately

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Fire and Life Safety

Environmental Health and Safety

Laboratory Fire Safety

• Flammable substances are among the most common hazardous materials found in laboratories
• Avoid accumulation of vapors and control sources of ignition
• Requires knowledge of flammability characteristics of materials encountered in the laboratory

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Properties of Flammable/Combustible Liquids

• Flash point - lowest temperature at which a liquid gives off vapor in sufficient concentration to form an ignitable mixture with air
• Many common laboratory solvents and chemicals have flash points below common room temperature
• Acetone, benzene, ethanol, methanol
• It is actually the vapor, not the liquid, which burns
• The rate at which different liquids produce flammable vapors depends on their vapor pressure

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Flammable/Combustible Liquids

• Flammable liquids
• Flash point below 100 °F (37.7 °C)
• Vapor pressure not exceeding 40 psi (276 kPa)
• Class IA: flash points below 73°F (22.8°C) and boiling point below 100°F (37.8°C)
• Class IB: flash points below 73°F (22.8°C) and boiling point at or above 100°F (37.8°C)
• Class IC: flash point at or above 73°F (22.8°C) and below 100°F (37.8°C)
• Combustible liquids
• Flash point ≥ 100°F (37.7 °C)
• Class II: flash points at or above 100°F (37.8°C) and below 140°F (60°C)
• Class III: flash points at or above 140°F (60°C)

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Properties of Flammable/Combustible Solids

• Flammable Solid (Class 4.1)
• Self-reactive materials that are thermally unstable and can undergo a strongly exothermic decomposition even without participation of oxygen; and desensitized explosives
• Spontaneously Combustible Material (Class 4.2)
• Pyrophoric (air-reactive) materials or self-heating materials, likely to self-heat when in contact with air and without energy supply
• Dangerous when wet material (Class 4.3)
• Liable to spontaneously combust or give off flammable/toxic gas when in contact with water

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Properties of Flammable/Combustible Gases

• Flammable gases (Class 2.1)
• Gases at 20° C (68° F) or less and 101.3 kPa (14.7 psi) of pressure, AND
• Are ignitable at 101.3 kPa (14.7 psi) when in a mixture of 13% or less by volume with air; OR
• Have a flammable range at 101.3 kPa (14.7 psi) with air of at least 12% regardless of the lower limit.

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Ignitability

• Auto-ignition temperature for solid, liquid or gas
• Minimum temperature required to initiate self-sustained combustion independent of the heat source.
• A steam line or a glowing light bulb may ignite carbon disulfide (ignition temperature 80°C [176°F]). Diethyl ether (ignition temperature 160°C [320°F]) can be ignited by the surface of a hot plate. Silane gas (ignition temperature 21°C [69.8 °F]) can spontaneously ignite at or near room temperature
• Spontaneous ignition or combustion
• A substance reaches its ignition temperature without the application of external heat.
• Consider the possibility of spontaneous combustion, especially when materials are stored or disposed.
• Materials susceptible to spontaneous combustion include organic materials mixed with strong oxidizing agents (such as nitric acid, chlorates, permanganates, peroxides and persulfates), the alkali metals (lithium, sodium, potassium, rubidium, and cesium), finely divided metal powders, and phosphorus.

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Sources of Ignition

• Potential sources of spark, flame, or heat in laboratories can ignite flammable substances
• Consider open flames, static electricity, lit matches and hot surfaces.
• Vapors of flammable liquids are heavier than air, and can travel considerable distances.
• Flammable vapors from substantial sources such as spills can descend into stairwells and elevator shafts and ignite on a lower story. If the path of vapor is continuous, the flame can propagate itself from the point of ignition back to its source.
• Properly bond and ground all metal lines and vessels dispensing flammable substances to discharge static electricity. When nonmetallic containers (especially plastic) are used, the bonding can be made to the liquid rather than to the container

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Flammable Substance Handling�Best Practices

• Handle flammable substances only in areas free of ignition sources
• Do not heat flammable substances with an open flame
• When you transfer flammable liquids in metal equipment, avoid static-generated sparks by bonding, and the use of ground straps
• Ventilation is one of the most effective ways to prevent the formation of flammable mixtures. Use fume hood when you handle appreciable quantities of flammable substances
• Containers of flammable liquids shall not be drawn from or filled within buildings without provisions to prevent the accumulation of flammable vapors in hazardous concentrations

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Flammable Substance Storage�Best Practices

• Store flammable and combustible liquids only in approved containers
• Flammables stored in the open in the laboratory work area shall be kept to a minimum
• Safety refrigerators are recommended and should be labeled for flammable material storage
• Domestic refrigerators in labs must have signage which states that no flammable storage is permitted
• Do not locate flammable storage cabinets near an exit or in egress pathways
• Keep flammable liquids away from heat and direct sunlight

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Flammable/Combustible Storage

• Maximum allowable size of containers for flammable and combustible liquids are set by NFPA

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OSHA Fire Guidance

IF clothing catches fire……..

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Types of Fire Extinguishers

• Most chemical laboratory fire hazards require multipurpose dry chemical extinguishers (ABC) located in hallways.
• “Gas” extinguishers containing carbon dioxide (CO2) offer a first defense against flammable liquids or electrical fires without leaving a powder residue that could harm electronic equipment.
• Class D extinguishers are also located in select labs, they can be identified by label and color (yellow).
• EHS coordinates monthly checks of all fire extinguishers. Please report any problems or missing extinguishers to ehs@nccu.edu.

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Means of Egress Components

Both OSHA and Fire Code provide requirements regarding the maintenance of egress routes and access. The maintenance of these routes is essential as they provide the means by which occupants can safely exit in the event of a fire or other emergency

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Egress routes are divided into three sections, exit access, exit, and exit discharge.

- An exit access is a portion of a building (room, hallway, aisle, etc.) that leads to an exit route

- An exit is a protected (fire rated) pathway by which occupants may move to an exit discharge

- An exit discharge is a way by which occupants leave an exit, leading from the building to a public way

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All exit components (access, route, and discharge) must be maintained so as to permit their immediate use by building occupants in the event of an emergency.

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Mean of Egress Maintenance

Exits and the entries to exits will be marked by overhead, illuminated exit signs and in some cases emergency lighting

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Exit access (hallways, aisles, etc.) that lead to the protected exit are required to be kept clear at all times – their width is determined by both OSHA and NC Fire Code

• OSHA 1910.36(g)(2) indicates a minimum width of 28”
• NC Fire Code (Section 1018.3) sets additional requirements – if fewer than 50 occupants, width must be at least 36”

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Items must not be stored within either the exit access or exit as such storage may reduce the component to below the required width – combustible items may not be stored in any quantity within an exit

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Storage Height and Contents

Care must be taken when storing items on shelving within laboratories. Items stored at too great of a height are not only difficult to access but may also impede the operation of the automatic sprinkler system in the building

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Generally, items may not be stored within 18” of the sprinkler head (NFPA 13 10.2.8.2)

• One exception – items stored on shelving against walls may exceed the 18” clearance provided they do not impede sprinkler operation

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In a non-sprinklered building, storage must not be within 24” of the ceiling (NCFC Section 315)

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This form is completed and posted near laboratory exit doors for awareness of lab specific procedures.

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Electrical Safety

• Periodic inspections should check that
• All sources with electrical potential have proper grounding or circuit breaking
• Cords and plugs are in a safe condition
• Multi-outlet power strips may be used for computers but NOT for lab equipment
• Possible electrical hazards include
• Spliced or worn cords
• Cords with missing ground pins
• Cords near hot plates or flames
• Cords used near water that are not protected with ground fault circuit interrupter

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Control of Hazardous Energy standard 29 CFR 1910.147

• Also referred to as lockout/tagout
• Procedures to shut down equipment or isolate it from its energy source to prevent injury while maintenance and servicing activities are being performed.
• For awareness – never remove a lock, tag or other device placed on equipment to prevent energization that you did not place on the equipment

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Additional Training Opportunities

Environmental Health and Safety

NCCU Safety Materials/Trainings

• Compressed Gas Safety Training (required initial training)
• Ergonomics Information Sheet
• Electrical Safety Sheet
• Formaldehyde Safety Training (required initial training)
• Exposure Control (Bloodborne Pathogens) Training (required initial and annual training)
• NCCU Autoclave Safety Training (required initial training)

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• 2021 Biosafety and Chemical Safety Manuals & Training

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Safety Training at CDC TRAIN

• Requires free account
• Provides certificate of completion

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Questions

Department of Environmental Health and Safety

(919) 530-7125

ehs@nccu.edu

EHS Website

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To complete this training you are required to pass a short quiz with a score of ≥ 80%

Click here to begin

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