Hands-On Laboratory Safety Training and Biosafety Training

Transcription

1 Hands-On Laboratory Safety Training and Biosafety Training Name: NetID: PI/Group: Date:

2

3 Station 1: Hazard Information The following questions refer to the University of Nevada, Reno Chemical Hygiene Plan (CHP). The most recent version of the Chemical Hygiene Plan can be found online at You may also wish to keep a physical copy of the Chemical Hygiene Plan in your laboratory and update it annually. 1. Does this laboratory have a hard copy of the Chemical Hygiene Plan? If so, where is it? 2. Find the Chemical Hygiene Plan on the EH&S website ( On which webpage did you find it? 3. On what page(s) of the CHP can you find information on the selection of chemically resistant gloves? 4. According to the CHP, how often should peroxide formers be tested for organic peroxides? 5. How many flammable gas cylinders may be legally stored in a lab area without fire sprinklers? 6. How often should laboratory groups perform a self-assessment of their research space? (Hint: Chapter 2) The following questions refer to information that can commonly be found on Safety Data Sheets (SDS). You may find Safety Data Sheets on manufacturer or supplier websites, via subscription services, or by using the EH&S Safety Data Sheet (SDS) Retrieval System. Physical copies of Safety Data Sheets (SDS) for highly hazardous or frequently used chemicals should be kept in the laboratory where they are stored. 7. You have a 2L bottle of Anhydrous Benzene from Sigma-Aldrich (Prod. # ). Use the manufacturer s website ( to locate the SDS and list two Hazard Statements found in the HAZARDS IDENTIFICATION section. 8. You find an old 100mL plastic bottle with a label that has been worn off in your laboratory. You want to dispose of it, but you don t know the contents. You can make out the product number and can read the word Plant in the product name. Use the EH&S SDS Database ( to find the chemical contents and list them here. 9. While on the University network, you also have access to the Canadian Centre for Occupational Health and Safety s SDS web resource a link to which can be found on the EH&S website ( Use the CCINFOWeb service to find an SDS for Aniline. What is the melting point? What is the physical state at room temperature? 10. Imagine there is an earthquake that knocks out the power to campus. During the tremors, a bottle falls off a shelf and the lid comes off. While you are doing your best to Drop, Cover, and Hold On, you kneel in the resulting puddle. It soaks through your pants and covers a large portion of your leg in a colorless liquid. You turn the bottle over and read the label: HYDROFLUORIC ACID, 48%. Assuming your lab is properly equipped for such a situation, what first aid measures need to be taken? REMEMBER: the power is out! No computers! When you are finished with Station 1: Hazard Information, please close any browser windows you have opened and return any items you may have moved to their original locations. Station 1: Hazard Information Page 1 of 1

4 Station 2: Laboratory Ventilation There are two fume hoods, start this section in Hood #1 which is cluttered with objects. Perform the following operations and answer the questions. 1. When was the last test on the hood performed? 2. What was the average face velocity in feet per minute (FPM) at the last test? (Note: the average face velocity range that is recommended by EH&S is FPM.) Lower the glass sash almost all the way down, allowing for a couple of inches of a gap for air flow. Position the Dragon Puffer hand held smoke generator approximately inches away from the front of the hood but at the same level as the gap between the sash and surface of the hood. Squeeze and hold the handle on the Dragon puffer to generate smoke. Slowly and gently sweep the generator across the width of the hood once or twice. 3. What happens to the smoke? Lift the sash to match the height of the red arrow on the testing sticker, move the smoke generator to a height halfway between the benchtop and the sash, keeping it about 10 inches from the face of the hood. Gently pass the Dragon Puffer from side-to-side. 4. What happens to the smoke? While keeping the smoke generator in the same position as the previous step, lift up the sash as high as it will go. 5. What happens to the smoke with the sash all the way up vs. at the height of the test sticker? Keep the sash all the way up, move the smoke generator down near the bottom of the hood and gently sweep it back and forth. Move the smoke generator closer to the front of the hood, within an inch or so, and gently move the smoke generator in front of and around the objects in the front of the hood. 6. What happens to the smoke around objects? (Note: when using equipment in a fume hood, it should be placed 4 6 inches back from the opening.) Move to Hood #2, the cleaner, clutter-free fume hood. Perform the following operation while demonstrating good technique in working inside a lab fume hood while using an appropriate sash height and working distance from the face for the individual performing the manipulations. While one person is measuring out materials, another team member can generate smoke from about 10 inches away around the person s arms and all observe the behavior. Measure out 1 teaspoon (approximately 10 grams) of citric acid, monohydrate and transfer it to the 500 ml Erlenmeyer flask. Measure out 200 ml of the hexanes into a graduated cylinder and transfer it to a 500 ml Erlenmeyer flask. Stir for 15 seconds with the magnetic stir bar and stir plate. Station 2: Laboratory Ventilation Page 1 of 2

5 Station 2: Laboratory Ventilation Transfer the contents to the provided waste container. Use the acetone rinse bottle to rinse the residual chemicals from the Erlenmeyer flask into the waste bottle. Ensure the waste bottle is properly left for storage. Fill out the sample waste log below. (Note: be sure to include everything that was added to the waste container and the appropriate quantities.) Clean up any liquid or solid spills in the hood. Close the fume hood sash, leaving a small gap at the bottom. (Note: always close fume hood sashes when not actively working in the hood.) 7. What did the smoke do while you were measuring out materials? Sample Waste Log Sheet Container # CHEMICAL NAME AMOUNT Station 2: Laboratory Ventilation Page 2 of 2

6 Station 3: Laboratory Operations You will be conducting a self-assessment of this training laboratory. Please identify as many safety deficiencies as you can by marking the appropriate shaded box. Items that have been pre-marked as NA do not apply to this space, although they may apply to your actual lab. If you find a safety deficiency that is not listed here, you may record it on the bottom of the next page. Please do not attempt to correct any identified findings so that other groups may find them as well. LABORATORY MANAGEMENT 1. Laboratory personnel are familiar with, and know how to access, the chemical hygiene plan, biosafety manual, and radiation safety manual. Yes No N/A 2. Written SOPs have been developed to cover specific laboratory operations X 3. Laboratory-specific safety training has been provided and documented X 4. SDSs and other hazard information are available and personnel know how to access them. 5. A current inventory of chemical, biological, and radiological hazards is readily available. 6. An approval or review process is used prior to the introduction of significant new risks to the laboratory. GENERAL LABORATORY SAFETY X X 7. Lab emergency contact information is posted at lab entrance. 8. Warning signs are posted to designate specific hazards. 9. Aisles are clear and without tripping hazards. 10. All exits are free and unobstructed. 11. Appropriate safety eyewear, gloves, lab coats, and other personal protective equipment is available and used. CHEMICAL LABELING AND STORAGE 12. Chemical containers are labeled with the content identity and hazard warnings. 13. Chemicals are segregated by major hazard categories and incompatibles are separated. 14. Peroxide forming chemicals are dated and tested at least annually. 15. Flammable and combustible liquids storage volumes are minimized and stored in flammable storage cabinets or safety cans. 16. Compressed gas cylinders are properly secured. 17. Compressed fuel gases are separated from oxidizing gases. 18. Flammable and toxic gases are stored and a well-ventilated area with highly toxic gases stored in a ventilated enclosure X Station 3: Laboratory Operations Page 1 of 2

7 Station 3: Laboratory Operations SAFETY EQUIPMENT Yes No N/A 19. An operable eyewash and emergency shower is readily available. 20. Fire extinguishers are available, unobstructed, and operational. 21. Chemical fume hoods are available, operational, and annual inspection is current. 22. Biological safety cabinets are available, operational, and annual inspection is current. CHEMICAL WASTE 23. Chemical waste containers are labeled and lids are closed. 24. Chemical waste is segregated by hazard class. 25. Storage of waste is minimized and full waste containers are not allowed to accumulate. 26. Biohazardous waste containers display the universal biohazard warning label. 27. Sharps are collected in puncture-proof, leak-proof containers displaying the biohazard warning label. 28. Extension cords are not used as permanent wiring. 29. Electrical cords are not frayed or damaged. ELECTRICAL SAFETY 30. Electrical cords contain a ground wire (three-prong plug) X 31. High voltages are shielded and adequately marked and labeled X EMERGENCY RESPONSE 32. Personnel have been trained to properly respond to spill, fire, and other incident scenarios X 33. Chemical spill cleanup materials are available. 34. Biological disinfectant and spill cleanup materials are available. 35. First aid kit is available and appropriately stocked X 36. Emergency phone numbers are posted by phone X Additional deficiencies identified: (This self-assessment checklist can be found in the appendix of the UNR Chemical Hygiene Plan. You may use it as-is while conducting regular self-assessments of your lab, or you may adapt it as you see fit.) Station 3: Laboratory Operations Page 2 of 2

8 Station 4: Incident Response At this station, you will participate in a chemical spill cleanup drill in which you will be expected to isolate the spill area and take appropriate measures based on the type and quantity of material spilled. A low-risk spill, where laboratory personnel fully understand the hazards and risks involved, will be presented. In order to be classified as a low-risk spill, adequate spill control materials are at hand; proper personal protective equipment is available; and workers have been trained in spill cleanup procedures. As a guideline, a low-risk spill can be thought of as 1 L or less of flammable liquid, concentrated acid or base, or 100 ml or less of a particularly hazardous substance. A clearly marked container of flammable liquid has spilled and/or is leaking. Respond to the spill by demonstrating the following techniques: Isolate the area and communicate the spill to others Shut off nearby ignition sources (if applicable) Wear appropriate PPE Stop the flow of the leaking chemical (if safe to do so) Prevent the spill from reaching drains Quickly encapsulate and/or absorb the spill using available materials Transfer chemicals from damaged/leaking containers (if possible) Thoroughly clean the area with detergent and water Prepare all cleanup materials for disposal (via submission to EH&S for pick-up) 1. Describe your process of cleaning up the spill at this station. 2. How would the cleanup process change if the waste was corrosive instead of flammable? 3. How would the cleanup process change if the identity of the spilled chemical was unknown? Station 4: Incident Response Page 1 of 1

9 Station 5: Biosafety Treat the liquid labeled as broth culture as if it contains a pathogenic bacterium that requires biosafety level 2 (BSL-2) containment procedures. Your group will perform steps 1 10 listed below. All members of the group are expected to participate throughout the exercise. As much as possible, have different people perform the following steps: decontamination of biosafety cabinet before you begin work (step 2) set up of biosafety cabinet and perform manipulations (steps 3 4) decontamination and removal of an item from biosafety cabinet (step 5) cleanup of a small spill inside the biosafety cabinet (step 6) cleanup of biosafety cabinet after work is finished (steps 7 10) Prepare to Work in Biosafety Cabinet 1. Briefly discuss what personal protective equipment (PPE) you should wear to conduct work at BSL-2. Put on (don) all necessary PPE. 2. Turn on the biosafety cabinet blower and then decontaminate the interior surfaces of the cabinet by wiping surfaces with 70% ethanol. 3. Place the materials listed below in the biosafety cabinet in a manner that will allow you to work from clean to dirty. Tube rack with 50 ml conical tube labeled diluted culture broth and 14 ml conical tube labeled culture broth Micropipette (with tips) Spray bottle labeled 70% ethanol 50 ml conical tube labeled Glo-Germ solution Handheld UV light Small plastic bucket lined with biohazardous waste bag Tongs Approximately 6 inch strip of autoclave tape o (stick one end of tape to interior side of cabinet so that it can be easily removed) Perform the Following Manipulations 4. Using the micropipette, transfer 0.25 ml of liquid from the 14 ml conical tube labeled culture broth to the 50 ml conical tube labeled diluted culture broth. Place the used pipette tip in the biohazardous waste container located in the biosafety cabinet. Place the cap on the 50 ml conical tube and gently swirl to mix contents. 5. Using 70% ethanol, decontaminate the exterior surface of the 50 ml conical tube labeled diluted culture broth. Spray gloves with 70% ethanol and rub gloved hands together. Remove 50 ml conical tube from the cabinet as if it will be used for an additional procedure at another location. 6. Simulate a small spill of the bacterial culture by pouring 1 2 ml of Glo Germ solution from a height of ~12 inches onto the working surface of the biosafety cabinet. Use handheld UV light to visualize spill initially, and then as the cleanup progresses. Clean up the liquid as if it were an actual spill of the bacterial culture. Note: You do not need to allow contact time between the spill and the disinfectant as you normally would. a. Begin cleanup by gently placing absorbent towels on the spill and then saturating the towels with 70% ethanol. Using tongs, place absorbent towels in biohazardous waste. b. Spray entire affected area with 70% ethanol and wipe area with absorbent towels. Place used towels in biohazardous waste. c. Spray gloves with 70% ethanol and rub gloved hands together. Station 5: Biosafety Page 1 of 2

10 Station 5: Biosafety Clean Up the Cabinet 7. Assume all other items in the biosafety cabinet except for the biohazardous waste container have been decontaminated and remove all items except for the biohazardous waste container from the cabinet. Note: Normally each item would be individually surface decontaminated prior to removal. 8. Remove the biohazardous waste bag by gently unrolling the bag from around the top of the waste container. Close the top of bag, forming a loose neck near the top, and wrap the approximately 6 inch strip of autoclave tape loosely around the neck. Pull the bag out of the biohazardous waste container, decontaminate the exterior of the bag using 70% ethanol, and then remove it from the biosafety cabinet and place it into the floor-standing biohazardous waste container. Note: Normally gloves would be sprayed with 70% ethanol before removing them from the cabinet. 9. Assume that the biohazardous waste container has been decontaminated and remove the biohazardous waste container from the cabinet. 10. Close the sash on the biosafety cabinet and turn off fan and light. Remove your gloves by turning each glove inside out starting at the cuff and rolling them over your hand. Avoid touching the exterior of your gloves or your skin. Place the gloves in the floor-standing biohazardous waste container. Questions 1. At a minimum, when should routine decontamination of the biosafety cabinet be performed? 2. 70% ethanol is used as a chemical disinfectant in this exercise. Should 70% ethanol always be used as a disinfectant when working with pathogens? 3. What should you do if you know that you got some of the broth culture on your gloved hand while you are working in the biosafety cabinet? Station 5: Biosafety Page 2 of 2