Using smaller quantities of hazardous chemicals or substituting a less hazardous chemical reduces the risk of serious exposure or spill. See Chapter VI, Hazardous Waste Directory for additional waste reduction information. When planning your work, consider the following possibilities:
- Substituting less hazardous chemicals;
- Using less;
- Ordering only what is needed; and
- Sharing chemicals when possible.
Use the safety equipment and personal protective equipment (PPE) recommended in Section 6, Personal Protective Equipment, or listed on container labels or MSDSs for a particular material or procedure.
3.2.1 Use Recommended Engineering Controls
If the release of a hazardous vapor, mist, gas, or dust is possible, perform the work using the appropriate engineering control, such as a chemical fume hood, glove box, or vented biosafety cabinet. Take the time to label temporary containers and inspect manufacturers' labels for thoroughness and accuracy. See Section 5.1 for details on labels.
3.2.2 Know the Hazard
Review the hazards of the chemicals before using them. Review the Material Safety Data Sheet and this chapter for safe handling procedures and PPE recommendations. Be prepared for a spill or an exposure involving the hazardous chemical. Know the location of the nearest eyewash and emergency shower.
3.2.3 Keep Halls Clear
Keep hallways outside the lab and routes of egress within the lab clear of furniture and equipment. Make sure access to the fire extinguisher and emergency eyewash and shower are unobstructed.
3.2.4 Unattended Operations
Experiments involving heat generating devices must never be left unattended. For other experiments left unattended, plan for interruption in utility services, such as electricity, cooling water, and gas. Place a sign near operations warning others of potential hazards and list emergency procedures to follow. Whenever possible, have someone check operations periodically.
3.2.5 Plan Carefully and Anticipate New Hazards
At the beginning of an extended project, formally analyze the procedures for possible hazards and consider the consequences. Ask a colleague to review the hazard analysis.
3.2.6 Do Not Work Alone
Do not work alone if your work requires the use of hazardous materials or hazardous processes. At a minimum, a second person should be aware of an individual working alone in the lab and arrangements should be made for periodic checks. Excessively long work hours increase the likelihood of mistakes and accidents due to fatigue.
3.2.7 Report Spills to EH&S
Report all spills, accidents and injuries to EH&S and complete an Accident/Illness Report (AIR) Form. See Chapter I, Section 3.1 for details on accident and illness reporting. After hours contact EH&S through Security.
Follow good housekeeping practices. Maintain work areas in an orderly fashion. Avoid accumulation of combustible materials. Cluttered areas increase the likelihood of accidents and injuries.
3.2.8 Do Not Rely on Odor as an Indicator of Exposure
The absence of odor is not a reliable guide to a safe concentration of airborne chemical in the lab. Concentrations detectable by odor vary according to the chemical and the ability of the individual to smell the chemical. Never rely on odor to determine exposure hazard.
3.2.9 Exposure Monitoring
If you are concerned about your chemical exposure or are experiencing symptoms associated with exposure to a chemical, contact EH&S for an exposure evaluation.
3.2.10 Food and Drink Policy
Never smell or taste chemicals to identify them. Wash your hands immediately after using any hazardous material and before leaving the lab. Never pipette or siphon liquids by mouth.
Do not eat, drink, or apply cosmetics in the lab. Do not store food or drinks in a lab refrigerator or in a cold room.
3.2.11 Prevent Chemical Releases in Cold Rooms
Do not store chemicals in cold rooms. Take all precautions to prevent material releases in cold rooms. Most cold rooms do not have ventilation; some have very little. Therefore, chemical vapors or fumes will not be diluted, which could cause an exposure hazard. Do not place dry ice in cold rooms because, as a simple asphyxiant, it displaces oxygen. Avoid storage of cellulose materials such as paper and cardboard to prevent fungal growth. For example, use plastic tubs instead of cardboard boxes.
3.2.12 Explosion Shielding
Use an explosion shield or other protective enclosure if there is a possibility of a violent reaction. Do not overlook the possibility that scaling up or heating a process will change the safety parameters.
3.2.13 Vent Apparatus
Vent equipment or containers which discharge vapors (vacuum pumps, distillation columns) into chemical fume hoods or through appropriate filters.
3.3 Centrifuge Safety
Centrifuges come in three general classes: low speed (up to about 5000 rpm), high speed (up to about 25,000 rpm), and ultracentrifuges (up to 100,000 rpm). Rotors on centrifuge and ultracentrifuge units are subjected to powerful mechanical stresses that may cause metal fatigue over time. This can lead to rotor failure presenting two serious hazards: mechanical failure and dispersion of aerosols.
Approximately 90% of rotor incidents are due to user error, primarily from failure to put the lid on the rotor, failure to secure the lid, and failure to properly secure the rotor to the drive.
3.3.1 Safety Procedures for Centrifugation
- The PI, lab supervisor, and LSC must train each operator on proper operating procedures. Each user must review the manufacturer's operating manual.
- Use only rotors compatible with the centrifuge. Check the expiration date for ultracentrifuge rotors.
- Check tubes, bottles, and rotors for cracks and deformities before each use.
- Make sure that the rotor, tubes, and spindle are dry and clean.
- Examine O-rings and replace if worn, cracked, or missing.
- Never overfill centrifuge tubes (don't exceed 3/4 full).
- Always cap tubes before centrifugation.
- Always balance buckets, tubes, and rotors properly.
- Check that the rotor is seated on the drive correctly, close the lid on the centrifuge, and secure it.
- When using swinging bucket rotors, make sure that all buckets are hooked correctly and move freely.
- Maintain a log to keep track of the number of runs or hours of use (see log info below).
- Close lids at all times during operation. Never open a centrifuge until the rotor has stopped.
- Do not exceed safe rotor speed.
- The operator should not leave the centrifuge until full operating speed is attained and the machine appears to be running safely without vibration.
- Stop the centrifuge immediately if an unusual condition (noise or vibration) begins, and check load balances.
- Allow the centrifuge to come to a complete stop before opening.
- Wear a new pair of outer gloves to remove rotor and samples.
- Check inside of centrifuge for possible spills and leaks, and clean centrifuge and rotor thoroughly, if necessary.
- Wash hands after removing gloves.
3.3.2 Centrifuging Infectious Materials or Human Samples
Follow safety procedures above, plus:
- Avoid the use of celluloid tubes with biohazards. If celluloid tubes must be used, an appropriate chemical disinfectant must be used to decontaminate them.
- Always use sealed safety cups, safety buckets, or sealed rotors with O-rings as secondary containment.
- Always wear gloves when handling tubes or rotors.
- Place a biohazard label on the centrifuge.
- Fill centrifuge tubes, load into rotors, remove from rotors, and open tubes within a biological safety cabinet whenever possible.
- Wipe exterior of tubes or bottles with disinfectant prior to loading into rotor or bucket. Seal rotor or bucket, remove outer gloves, and transport to the centrifuge.
- Wait at least 10 minutes after the run to allow aerosols to settle before opening the centrifuge. Check for possible spills or leaks. For spills of infectious materials transmitted by inhalation, see emergency procedures below.
- If tube breakage occurs, decontaminate centrifuge interior, safety cups or buckets, and rotors. See emergency procedures below.
- Include centrifugation procedure and decontamination plan in lab SOPs.
3.3.3 Emergency Procedures
For spills of infectious materials transmittedby inhalation:
- If a spill has occurred in the centrifuge outside of a biosafety cabinet, hold your breath, close the centrifuge lid, turn centrifuge off, and immediately leave the lab, inform supervisor and call EH&S.
- Notify others to evacuate the lab, close the door, post a biohazard spill sign at the lab door.
- Remove any contaminated protective clothing and place in a biohazard bag. Wash hands and any exposed skin surfaces with soap and water. Seek medical attention as necessary.
- Report spills to EH&S.
For malfunction, rotor failure, or tube breakage of materials not transmitted by inhalation:
- If a centrifuge malfunctions while in operation, turn it off immediately and unplug.
- If tube breakage occurs, turn centrifuge off immediately. Leave for 30 minutes to reduce the risk of aerosols. The operator should wear proper gloves, remove debris, clean and disinfect centrifuge interior, rotors, and safety cups or buckets following the manufacturer's instructions.
3.3.4 Centrifuge Maintenance
Moisture, chemicals, strong cleaning agents, and other substances can promote corrosion of centrifuge parts and cause centrifuge failure. The following are general maintenance recommendations:
- Follow manufacturer's instructions for maintenance and cleaning.
- Keep the centrifuge clean and dry.
- Clean all spills immediately and decontaminate the rotor after use with biological or radioactive materials.
- Clean rotors and cups with non-corrosive detergents (mild detergent and distilled water are recommended), then dry the surface thoroughly. Do not use bleach, as it tends to corrode parts.
- Never clean rotors and associated parts with abrasive wire brushes.
- Store the rotor upside down in a dry place, with lids or plugs removed to prevent condensation.
- Remove adapters after use and inspect for corrosion.
- Inspect rotor regularly. Remove rotors that show any signs of defect from use, and report them to a manufacturer's representative for inspection.
- Know your rotors and maintain a log book:
a. Rotors (high speed and ultra) of unknown history may not be used.
b. A complete and comprehensive rotor log is to be kept for every high-speed and ultracentrifuge rotor, and should include rotor serial number, user names, run dates, durations, speeds, total rotor revolutions, and any notes on rotor condition.
c. Rotor manufacture date and the date of stress testing (magnaflux or other professionally recognized analysis), if applicable, shall be recorded and kept with the rotor log.
d. Rotors are derated (their speed is lowered permanently or they are retired) after the manufacturers' recommended revolutions or years of service, whichever comes first, except where an annual stress test (magnaflux or other professionally recognized analysis) proves an absence of structural flaws.
e. Track and discard rotors according to the manufacturer's recommended schedule.
3.4 Chemical Disposal and Storage
3.4.1 Chemical Disposal
Do not mix hazardous wastes. Waste streams must be segregated for safe and cost-effective disposal. Correct disposal instructions are indicated for each chemical in Chapter VI, Hazardous Waste Directory. If you do not find the chemical listing in the directory, contact EH&S for disposal information.
3.4.2 Transporting Chemicals within the Fred Hutch Properties
Use safety carriers or secondary containers to transport dangerous chemicals (e.g., strong corrosives, solvents), even for short distances. To move several bottles at once, use a low cart with a substantial rim and segregate hazard classes with tubs. Do not carry hazardous chemicals in stairwells.
3.4.3 Emergency Equipment Checklist
The following emergency equipment must be located within or near the lab. Know the location and operation of the following:
- Dry chemical fire extinguisher;
- Emergency shower;
- Stocked first aid kit;
- Evacuation route map (posted); and
- Emergency response instructions (Fred Hutch Emergency Guide).
Contact EH&S if any of these items is missing.