Laboratory Safety

The following information has been collected and condensed from the EH&S website and other regulatory agencies and has been provided for reference and guidance to those working in ChBE research labs. The duties of various lab users are also provided below.

For general lab questions, please contact ChBE Safety Coordinator Ann Greco.

Principle Investigators (PIs)
  • Take a Hazardous Materials training class if you will be handling hazardous material/waste or managing people who do
  • Make sure lab users are up to date on their training
  • Delegate responsibility for safety to a lab Safety Proctor and hold him or her accountable
  • Apply appropriate penalties to protocol violators
  • Actually walk through the lab from time to time and make sure safety protocols are being followed!
  • A culture of safety starts from the top; emphasize it!
  • Make sure you know who is in the lab and what experiments they are running on a usual basis
  • Ensure that renovations and designs for new laboratory facilities incorporate required safety features
Lab Safety Proctors
  • Take a Hazardous Materials training class if you will be handling hazardous material/waste or managing people who do
  • Take the Gas Cylinder exam if you will be handling gas cylinders or liquid nitrogen dewars (please see Gas Cylinder tab for details)
  • Weekly: maintain SAAs, including packaging waste materials destined to be picked up by EHS
  • Bimonthly (every 2 months): check the status or operation of general safety equipment such as fire extinguishers, drench hoses, safety showers and eyewash stations
  • Post safety signs and instructions throughout the lab
  • Maintain a complete written current inventory of all chemicals, gases, biological, radioactive and other hazardous materials in the lab
  • Annually: conduct internal safety audit and recommend improvements to PI
  • Monitor storage, labeling and use of hazardous chemicals
  • Continuously monitor lab users for adherence to safety protocols
  • Exercise emergency authority to stop and prevent unsafe acts
  • Obtain statements of informed consent from all individuals where potentially hazardous materials are utilized
Graduate Students / Post Docs
  • Take a Hazardous Materials training class if you will be handling hazardous material/waste or managing people who do
  • Take the Gas Cylinder exam if you will be handling gas cylinders or liquid nitrogen dewars (please see Gas Cylinder tab for details)
  • Work in the hood when appropriate
  • Do not touch doorknobs or other inappropriate areas while wearing gloves
  • Familiarize yourself with the locations of all lab safety equipment including:
    • Routes of escape in case of fire or other emergencies
    • Fire extinguishers
    • First aid kits
    • Nearest available telephone
    • Emergency numbers (303-492-6666 for CU Police; 911 for emergencies)
  • Find the locations of SAAs and various disposal receptacles (for instance, sharps)
  • Familiarize yourself with the hazards and safety information of materials you are working with (MSDS)
  • Make sure you know any special safety information (monomer disposal, respiratory wear, etc.) that is associated with your lab or research
  • Always wear Personal Protective Equipment (PPE) in the lab (safety glasses, closed-toe shoes, gloves/lab coat if appropriate)
  • Make sure your undergrads have undergone the appropriate training for their work
  • Know what your undergrads are doing in the lab and supervise when necessary
Undergraduate Students
  • Take a Hazardous Materials training class if you will be handling hazardous material/waste or managing people who do
  • Take the Gas Cylinder exam if you will be handling gas cylinders or liquid nitrogen dewars (please see Gas Cylinder tab for details)
  • Know who’s who in the lab (Grad students, Post Docs, Safety Proctor)
  • Familiarize yourself with the locations of all lab safety equipment including:
    • Routes of escape in case of fire or other emergencies
    • Fire extinguishers
    • First aid kits
    • Nearest available telephone
    • Emergency numbers (303-492-6666 for CU Police; 911 for emergencies)
  • Find the locations of SAAs and various disposal receptacles (for instance, sharps)
  • Familiarize yourself with the hazards and safety information of materials you are working with (MSDS)
  • Make sure you know any special safety information (monomer disposal, respiratory wear, etc.) that is associated with your lab or research
  • Do not touch doorknobs or other inappropriate areas while wearing gloves
  • Always wear Personal Protective Equipment (PPE) in the lab (safety glasses, closed-toe shoes, gloves/lab coat if appropriate)
Hazardous Materials Training

If you handle hazardous materials, you must be formally trained on how to handle and appropriately dispose of these materials.

Environmental Health and Safety (EH&S) provides training courses each month. You can attend these courses for initial instruction or as a refresher course. The initial instruction must be completed within six months of beginning work with hazardous materials.

Gas Cylinders
Gas cylindersTraining for Transport of Compressed Gas Cylinders and Liquid Nitrogen Dewars

All faculty and students who intend to use compressed gas cylinders are required to pass a written exam.

First please review the presentation, "Safety Training for Transport of Compressed Gas Cylinders."

Then please download and complete the Gas Cylinder Safety Written Exam.

Once you have completed this exam, please return it to ChBE Safety Coordinator Ann Greco.

Fume Hoods

Use a chemical fume hood anytime your work involves:

  • Chemicals with a National Fire Protection Association (NFPA) Health rating of 3 or 4
  • Toxic volatile materials (chloroform, formaldehyde)
  • Flammable chemicals
  • Carcinogens or particularly hazardous substances
  • A procedure that may create an aerosol of a toxic substance
  • Reactive or explosive materials or chemicals that may spatter
  • Toxic gases (NH3, CO, F2, Cl2, H2S, NO2, etc.)
  • Odorous materials, both hazardous and non-hazardous

The sash is a blast shield, and provides primary containment in the event of fire. Your sash should be closed when you are not actively working at your hood. Moreover, you should be vigilant that your fellow lab occupants maintain closed sashes when they are not working at their hoods.

In cases where you are performing experiments that are suspected to present an explosion hazard, you should use a portable blast shield inside the hood in addition to a closed sash. You should also inform your fellow lab occupants when you are performing potentially hazardous procedures. Again, from the perspective of personal self interest, you should strive to maintain the sash at the lowest possible position at all times.

Chemical Handling
Chemical Properties

All chemicals are, to some degree, poisonous to the human body. Routes of entry include inhalation, skin and eye absorption, ingestion and injection. All personnel should be properly trained in use and application of MSDS - knowledge of properties, reactivities and compatibilities of chemical constituents, proper design and use of apparatus, engineering controls and correct PPE.

Dating Chemicals

All chemicals should be dated upon receipt and again upon opening. It is especially important that this procedure be done for all reactive and peroxide formers such as ethyl ether, tetrahydrofuran, etc. A first-in, first out (FIFO) inventory system should be adopted to control excess accumulation of chemicals and to prevent expired chemicals from automatically becoming regulated hazard wastes.

Reduce/Reuse/Recycle

All users of hazardous materials are required by EPA law to recycle chemicals, purchase less toxic materials, or use smaller quantities and design procedures that reduce the volume and concentration of hazardous materials used and waste generated.

Carrying/Transporting

Bottles, when carried, should always be supported on the bottom and never carried by the neck. Use a carrying device such as a rubber bucket which provides secondary spill containment and breakage protection. Whenever possible, protective coated chemical bottles and glassware should be used. A sturdy step stool or ladder should be used when obtaining chemicals from upper shelves that are out of reach. Transporting hazardous materials in vehicles involves extensive training for compliance with Federal, State, and local regulations. Contact EH&S for details before attempting transportation and to assure compliance with the law.

Toxics

Hydrofluoric acid, perchlorates, perchloric acid, radioactive materials, pyrophorics, gases and other extremely toxic, reactive, or potentially explosive materials should be handled under the direct supervision of the instructor or research staff and only after consultation with EH&S. Also, the use of perchloric acid may require a specially designed and designated fume hood. A special license must be obtained and a training course completed before any radioactive materials may be used.

Facility Closure

Areas where hazardous or radioactive materials are used or stored must be thoroughly decontaminated using EH&S approved methods prior to maintenance, renovation, reallocation of space, or closure. It is the responsibility of the supervisor, principal investigator and their department to arrange proper disposal of all hazardous materials prior to personnel relocations or facility closure.

Disposal

All chemicals, including those for disposal, must be clearly and completely labeled with full chemical names in English. This will aid emergency personnel, lab users (especially where changes of personnel and lab renovations have occurred), waste program personnel, and other building occupants to identify hazards and handle or dispose of chemicals properly.

Chemical Storage

Chemicals should be stored in approved closed containers and cabinets with secondary containment to prevent releases, separated by compatible hazard class (flammable/oxidizers/acids/bases/reactives) to avoid unwanted reactions and unnecessary exposure to occupants.

  • Chemicals must be stored in secured areas, i.e., not accessible to the general public.
  • Highly toxic and reactive materials need additional means of security such as lockable cabinets.
  • Whenever possible, protective coated chemical bottles and glassware should be purchased and used to reduce hazardous spills due to breakage.
  • It is important to note that certain hazardous materials are not permitted in many of our campus buildings, since the facilities are not constructed for high hazard use. Such materials include highly toxic gases, pyrophorics and highly reactive or unstable compounds. EH&S should be consulted before any of these substances are brought onto Campus.
  • Heavier items should always be stored closer to the ground.
  • Bottles of chemicals should never be stored on the floor.
  • Secondary containment can be provided by the use of plastic tubs or storage cabinets with containment features to prevent the spread of spilled or leaking chemicals.
  • Containment materials used should not be reactive with the chemicals stored in them.
Flammables

EH&S approved flammable liquid storage cabinets should have proper exhaust ventilation. Shelves and cabinets should be anchored solidly to the wall and safety lips should be installed along the front edges of exposed shelves to keep materials from falling. Flammable, volatile chemicals should be kept in a cool place, away from sources of heat and ignition.

Flammable Solvents

The total volume of flammable solvents in the laboratory should be limited to the amount needed for approximately one week of operations or the limit prescribed by NFPA (National Fire Protection Association), UBC (Uniform Building Code), and UFC (Uniform Fire Code), whichever is more restrictive.

Flammable Refrigerators

If flammables are stored in refrigerators/freezers, the units should be designed, manufactured and UL-approved to have spark-free interiors. Any refrigerator or freezer not designed for the storage of flammables needs to have "EXPLOSION HAZARD: Do Not Store Flammables in This Refrigerator" marked on the outside of the door.

Biological Material
Definitions

BIOLOGICAL WASTE (Infectious & Non-Infectious): Cultures, plates, media and other liquid or solid materials generated by Campus research and teaching laboratories, that contain or come in contact with living cells, body fluids, viruses, clinical materials, and other microorganisms.

INFECTIOUS WASTE: Biological waste that involves the presence of organisms containing recombinant DNA or any other organisms hazardous to human or animal health, including pathogens of sufficient virulence and quantity that exposure to the waste by a susceptible host could result in an infectious disease.

NON-INFECTIOUS: Plates, slides, culture vessels, and other biological or biomedical appearing materials generated by Campus laboratories, that do not meet the criteria of “infectious” (as defined above) or have been rendered non-infectious by chemical disinfection or autoclaving.

Introduction

The Institutional Biosafety Committee (IBC) requires that the following standard and special microbiological practices, physical containment or laboratory design, containment equipment, and training be implemented when research or teaching activities involve the use of biohazards, recombinant DNA molecules (rDNA), select agents, or bloodborne pathogens. These requirements include hygienic and operational practices that are critical in providing for a safe work environment and assuring a viable research product is produced. These practices are also necessary for minimizing and/or eliminating the risk of occupational exposure to infectious and potentially infectious substances.

Standard Practices

The first principle of containment is strict adherence to good microbiological practices. Consequently, all personnel directly or indirectly involved in experiments using microbial material shall receive adequate instruction. At a minimum, these instructions include training in aseptic techniques and in the biology of the organisms used in the experiments so that the potential biohazards can be understood and appreciated.

Any research group working with agents that are known or potential biohazards shall have an emergency plan that describes the procedures to be followed if an accident contaminates personnel or the environment. The Principal Investigator shall ensure that everyone in the laboratory is familiar with both the potential hazards of the work and the emergency plan. If a research group is working with a known pathogen for which there is an effective vaccine, the vaccine should be made available to all workers.

Biological Waste Generators Responsibilities
  • Determine if the waste in infectious. Render all infectious waste non-infectious using effective chemical disinfection methods or by autoclaving. Non-infectious liquid waste may be drain-disposed if it contains no other regulated chemicals or radioactive materials.
  • Place infectious waste that has not been chemically disinfected into non-leaking, heat resistant autoclave bags which have built-in sterilization indicators. Complete applicable portions of a non-biohazardous waste certification tag, check the "Autoclave" box and attach all three tag copies to the bag. Bring the bag of biohazardous waste to your designated autoclave area.
  • Biomedically appearing sharps (needles, syringes, blades, scalpels) whether they are contaminated or not, must be placed in puncture-proof, sealed containers and tagged for hazardous material/waste pick-up. Infectious sharps must be rendered non-infectious prior to submittal of the hazardous material/waste tag. Chemical disinfection is not effective for needles and small syringes. These types of infectious sharps must be autoclaved using a puncture-proof container that contains a built-in sterilization indicator or place autoclave tape on the container. Once the container has been autoclaved, make sure the autoclave indicator has changed to show that the sharps have been rendered non-infectious.
  • Seal biologically appearing non-infectious waste (No Sharps) inside a non-leaking autoclave bag, and complete and sign a non0biohazardous waste tag as described below, checking the box that indicates the material inside the bag is non-biohazardous. Attach the 3-part biowaste tag to the bag, remove the top (white) copy and put it in the designated pocket posted in the area near the autoclave. Leave the other two copies of the tag attached to the bag and place the bag into a designated "Certified Non-biohazardous Materials" receptacle. There is no need to autoclave or chemically disinfect this waste.
  • Seal chemically disinfected solid biowaste (No Sharps) inside a non-leaking autoclave bag and complete and sign a non-biohazardous waste tag as described below, indicating that the material inside the bag has been redered non-infectious by chemical treatment,. Attach the tag to the bag and put the top (white) copy into the designated pocket. Place the bag into a designated "Certified Non-biohazardous Materials" receptacle.
Biological Safety Cabinets

Biological safety cabinets referred to in this section are classified as Class I or Class II cabinets. For additional information on biological safety cabinets, please visit: CDC-NIH Selection, Installation and Use of Biological Safety Cabinets

Class I - The Class I BSC provides personnel and environmental protection, but no product protection. It is similar in air movement to a chemical fume hood, but has a HEPA filter in the exhaust system to protect the environment. In the Class I BSC, unfiltered room air is drawn across the work surface. Personnel protection is provided by this inward airflow as long as a minimum velocity of 75 linear feet per minute (lfpm) is maintained through the front opening. Class I BSCs are used specifically to enclose equipment (e.g., centrifuges, harvesting equipment or small fermenters), or procedures with potential to generate aerosols (e.g. cage dumping, culture aeration or tissue homogenation).

Class II - Class II BSCs are partial barrier systems that rely on the laminar movement of air to provide containment. If the air curtain is disrupted (e.g., movement of materials in and out of a cabinet, rapid or sweeping movement of the arms) the potential for contaminant release into the laboratory work environment is increased as is the risk of product contamination. The Class II (Types A1, A2, B1 and B2) BSCs provide personnel, environmental and product protection. Airflow is drawn into the front grille of the cabinet, providing personnel protection. In addition, the downward laminar flow of HEPA-filtered air provides product protection by minimizing the chance of cross-contamination across the work surface of the cabinet. Because cabinet exhaust air is passed through a certified HEPA filter, it is particulate-free (environmental protection), and may be recirculated to the laboratory or discharged from the building via a canopy connection.

Radiation
Radiation information

Radioactivity is defined as the spontaneous emission of radiation, generally alpha and beta particles, often accompanied by gamma rays, from the nucleus of an unstable atom. Radiation may be particles or photons emitted from an unstable radioactive atom as a result of radioactive decay. All of these types of radiation are represented at the University of Colorado. Each type of radiation has unique safety considerations and holding techniques that will be discussed.

Time, Distance, Shielding

Radiation may be reduced by taking advantage of time, distance, and shielding. By reducing the time spent working with radioactive materials and/or radiation producing machines, the dose received from the radiation is reduced.

Increasing the distance from a source also will reduce the dose because the intestate of radiation decreases as approximately 1/d2, where d is the distance from the source. For example, if the distance is doubled, the intensity is reduced to 1/4 of the original intensity. This is also known as the Inverse Square Law. Shielding can be very effective in reducing the dose received.

There are different types of shielding for different types of radiation. Use caution when selecting shielding to reduce the radiation dose. The dose may actually increase by selecting the wrong shielding. Verify radiation levels with a survey meter to ensure that appropriate and/or enough shielding has been used.

Storage

All freezers and other equipment used to store radioactive materials must have a Caution Radioactive Materials sign or label. Radioactive materials should be stored only in areas properly marked and approved for their use. Each laboratory must ensure security of radioactive materials and/or radiation-producing machines. This may require locking of laboratory doors or storage freezers/refrigerators depending on use and accessibility of the area.

Inventory

An inventory of all sealed or unsealed radioactive materials should be kept on the outside of the main storage freezer/refrigerator/area in each laboratory. Enter the date and the initials of the individual placing or disposing of an item being crossed off the inventory. The inventory sheet is collected to update the laboratory's possession levels.

It is a good practice to dispose of radioactive materials which are more than one or two years old, especially those bound to nuclide and proteins. Some bound radioactive materials and their chemical carriers have an effective "shelf-life" that may be exceeded.

Waste separation

Radioactive waste is separated into three types: solid, liquid, and scintillation vials. Each type has specifically designated waste containers. Containers are also provided for sharps, lead pigs, and any other unusual wastes. Empty lead pigs are stored separately and collected upon request for possible recycling. Unlike lead pigs, plastic pigs may be disposed in the appropriate solid waste container.

Radioactive waste is also segregated by half-life. There are three half-life categories designated by color.

The half-life categories are as follows:

  • Yellow: P-32, P-33, Rb-86 and other radionuclides with half-lives < 60 days
  • Orange: S-35, I-125 and other radionuclides with half-lives >60 days but < 90 days
  • Green: H-3 and C-14 and other radionuclides with half-lives > 90 days

Half-life categories are very important for waste minimization and decreasing disposal costs for the University.  Waste should be segregated by half-life category whenever possible and placed in the appropriately colored waste container. If waste is created containing two or more isotopes from different half-life categories, the waste should be disposed in the container for the longest lived isotope in the waste.

Mixed waste

There are three different types of radioactive waste created in a radiation laboratory: 1) purely radioactive, 2) mixed (radioactive and chemical), and 3) radioactive and biological. Proper handling of wastes is critical for appropriate transportation and disposal. When the containers are full, the laboratory submits a Radioactive Waste Pick-up Request Form.

Mixed waste (hazardous and radioactive) must comply with both radioactive and chemical regulations. Generation of mixed wastes should be avoided whenever possible. Disposal of this type of waste is very difficult and costly. Laboratories should actively seek ways to reduce the amount of mixed waste generated. Cost for mixed waste disposal may be re-charged to the laboratory.

Mixing biological wastes and radioactive material should be abided whenever possible. Any biological material must be rendered non-infectious using bleach or other disinfecting agent prior to disposal. When radioactive material is involved, use of an autoclave is NOT permitted. Once rendered non-infectious, this waste should be segregated from all other radioactive wastes. Do not use biohazard bags for radioactive materials.

EH&S Websites

Research Support 

EH&S Guidance Documents

Biological

Chemical

Disposal Requirements