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Laboratory Safety Manual

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Section 7: Safe Work Practices and Procedures

7I: Laboratory Equipment

Refrigerators and Freezers (top)

The potential hazards posed by laboratory refrigerators and freezers involve vapors from the contents, the possible presence of incompatible chemicals and spillage.

Only refrigerators and freezers specified for laboratory use should be utilized for the storage of chemicals. These refrigerators have been constructed with special design factors, such as heavy-duty cords and corrosion resistant interiors to help reduce the risk of fire or explosions in the lab.

Standard refrigerators have electrical fans and motors that make them potential ignition sources for flammable vapors. Do not store flammable liquids in a refrigerator unless it is approved for such storage. Flammable liquid-approved refrigerators are designed with spark-producing parts on the outside to avoid accidental ignition. If refrigeration is needed inside a flammable-storage room, you should use an explosion-proof refrigerator. 

Frost-free refrigerators should also be avoided. Many of them have a drain or tube or hole that carries water and possibly any spilled materials to an area near the compression, which may spark. Electric heaters used to defrost the freezing coils can also spark. 

Only chemicals should be stored in chemical storage refrigerators; lab refrigerators should not be used for food storage or preparation. Refrigerators should be labeled for their intended purpose; labels reading “No Food or Drink to be Stored in this Refrigerator” or “Refrigerator For Food Only” are available from EHS by calling 8-5294. 

All materials in refrigerators or freezers should be labeled with the contents, owner, date of acquisition or preparation and nature of any potential hazard. Since refrigerators are often used for storage of large quantities of small vials and test tubes, a reference to a list outside of the refrigerator could be used. Labels and ink used to identify materials in the refrigerators should be water-resistant. 

All containers should be sealed, preferably with a cap. Containers should be placed in secondary containers, or catch pans should be used. 

Loss of electrical power can produce extremely hazardous situations. Flammable or toxic vapors may be released from refrigerators and freezers as chemicals warm up and/or certain reactive materials may decompose energetically upon warming.  Proactive planning can avoid product loss and hazardous situations in event of an extended power outage. Dry ice or alternate power sources can be used to prevent refrigerator and freezer contents from warming.

Stirring and Mixing Devices (top)

The stirring and mixing devices commonly found in laboratories include stirring motors, magnetic stirrers, shakers, small pumps for fluids and rotary evaporators for solvent removal. These devices are typically used in laboratory operations that are performed in a hood, and it is important that they be operated in a way that precludes the generation of electrical sparks.

Only spark-free induction motors should be used in power stirring and mixing devices or any other rotating equipment used for laboratory operations. While the motors in most of the currently marketed stirring and mixing devices meet this criterion, their on-off switches and rheostat-type speed controls can produce an electrical spark because they have exposed electrical conductors. The speed of an induction motor operating under a load should not be controlled by a variable autotransformer. 

Because stirring and mixing devices, especially stirring motors and magnetic stirrers, are often operated for fairly long periods without constant attention, the consequences of stirrer failure, electrical overload or blockage of the motion of the stirring impeller should be considered.

Heating Devices (top)

Most labs use at least one type of heating device, such as ovens, hot plates, heating mantles and tapes, oil baths, salt baths, sand baths, air baths, hot-tube furnaces, hot-air guns and microwave ovens. Steam-heated devices are generally preferred whenever temperatures of 100o C or less are required because they do not present shock or spark risks and can be left unattended with assurance that their temperature will never exceed 100o C. Ensure the supply of water for steam generation is sufficient prior to leaving the reaction for any extended period of time.

A number of general precautions need to be taken when working with heating devices in the laboratory. When working with heating devices, consider the following: 

  • The actual heating element in any laboratory heating device should be enclosed in such a fashion as to prevent a laboratory worker or any metallic conductor from accidentally touching the wire carrying the electric current. 
  • Heating device becomes so worn or damaged that its heating element is exposed, the device should be either discarded or repaired before it is used again. 
  • Laboratory heating devices should be used with a variable autotransformer to control the input voltage by supplying some fraction of the total line voltage, typically 110 V.
  • The external cases of all variable autotransformers have perforations for cooling by ventilation and, therefore, should be located where water and other chemicals cannot be spilled onto them and where they will not be exposed to flammable liquids or vapors. 

Fail-safe devices can prevent fires or explosions that may arise if the temperature of a reaction increases significantly because of a change in line voltage, the accidental loss of reaction solvent or loss of cooling. Some devices will turn off the electric power if the temperature of the heating device exceeds some preset limit or if the flow of cooling water through a condenser is stopped owing to the loss of water pressure or loosening of the water supply hose to a condenser. 

Ovens

Electrically heated ovens are commonly used in the laboratory to remove water or other solvents from chemical samples and to dry laboratory glassware. Never use laboratory ovens for human food preparation.

  • Laboratory ovens should be constructed such that their heating elements and their temperature controls are physically separated from their interior atmospheres.
  • Laboratory ovens rarely have a provision for preventing the discharge of the substances volatilized in them. Connecting the oven vent directly to an exhaust system can reduce the possibility of substances escaping into the lab or an explosive concentration developing within the oven. 
  • Ovens should not be used to dry any chemical sample that might pose a hazard because of acute or chronic toxicity unless special precautions have been taken to ensure continuous venting of the atmosphere inside the oven. 
  • To avoid explosion, glassware that has been rinsed with an organic solvent should be rinsed again with distilled water before being dried in an oven.
  • Bimetallic strip thermometers are preferred for monitoring oven temperatures. Mercury thermometers should not be mounted through holes in the top of ovens so that the bulb hangs into the oven. Should a mercury thermometer be broken in an oven of any type, the oven should be closed and turned off immediately, and it should remain closed until cool. All mercury should be removed from the cold oven with the use of appropriate cleaning equipment and procedures in order to avoid mercury exposure.

Hot Plates

Laboratory hot plates are normally used for heating solutions to 100o C or above when inherently safer steam baths cannot be used. Any newly purchased hot plates should be designed in a way that avoids electrical sparks. However, many older hot plates pose an electrical spark hazard arising from either the on-off switch located on the hot plate, the bimetallic thermostat used to regulate the temperature or both. Laboratory workers should be warned of the spark hazard associated with older hot plates.

In addition to the spark hazard, old and corroded bimetallic thermostats in these devices can eventually fuse shut and deliver full, continuous current to a hot plate. 

  • Do not store volatile flammable materials near a hot plate 
  • Limit use of older hot plates for flammable materials. 
  • Check for corrosion of thermostats. Corroded bimetallic thermostats can be repaired or reconfigured to avoid spark hazards. Contact EHS for more info. 

Heating Mantles

Heating mantles are commonly used for heating round-bottomed flasks, reaction kettles and related reaction vessels. These mantles enclose a heating element in a series of layers of fiberglass cloth. As long as the fiberglass coating is not worn or broken, and as long as no water or other chemicals are spilled into the mantle, heating mantles pose no shock hazard.

  • Always use a heating mantle with a variable autotransformer to control the input voltage. Never plug them directly into a 110-V line.
  • Be careful not to exceed the input voltage recommended by the mantle manufacturer. Higher voltages will cause it to overheat, melt the fiberglass insulation and expose the bare heating element. 
  • If the heating mantle has an outer metal case that provides physical protection against damage to the fiberglass, it is good practice to ground the outer metal case to protect against an electric shock if the heating element inside the mantle shorts against the metal case. 
  • Some older equipment might have asbestos insulation rather than fiberglass. Contact EHS to replace the insulation and for proper disposal of the asbestos. 

Oil, Salt and Sand Baths

Electrically heated oil baths are often used to heat small or irregularly shaped vessels or when a stable heat source that can be maintained at a constant temperature is desired. Molten salt baths, like hot oil baths, offer the advantages of good heat transfer, commonly have a higher operating range (e.g., 200 to 425oC) and may have a high thermal stability (e.g., 540oC).There are several precautions to take when working with these types of heating devices:

  • Take care with hot oil baths not to generate smoke or have the oil burst into flames from overheating.
  • Always monitor oil baths by using a thermometer or other thermal sensing devices to ensure that its temperature does not exceed the flash point of the oil being used. 
  • Fit oil baths left unattended with thermal sensing devices that will turn off the electric power if the bath overheats. 
  • Mix oil baths well to ensure that there are no “hot spots” around the elements that take the surrounding oil to unacceptable temperatures. 
  • Contain heated oil in a vessel that can withstand an accidental strike by a hard object. 
  • Mount baths carefully on a stable horizontal support such as a laboratory jack that can be raised or lowered without danger of the bath tipping over. Iron rings are not acceptable supports for hot baths. 
  • Clamp equipment high enough above a hot bath that if the reaction begins to overheat, the bath can be lowered immediately and replaced with a cooling bath without having to readjust the equipment setup. 
  • Provide secondary containment in the event of a spill of hot oil. 
  • Wear heat-resistant gloves when handling a hot bath. 
  • The reaction container used in a molten salt bath must be able to withstand a very rapid heat-up to a temperature above the melting point of salt. 
  • Take care to keep salt baths dry since they are hygroscopic, which can cause hazardous popping and splattering if the absorbed water vaporizes during heat-up. 

Hot Air Baths and Tube Furnaces

Hot air baths are used in the lab as heating devices. Nitrogen is preferred for reactions involving flammable materials. Electrically heated air baths are frequently used to heat small or irregularly shaped vessels. One drawback of the hot air bath is that they have a low heat capacity. As a result, these baths normally have to be heated to 100oC or more above the target temperature. Tube furnaces are often used for high-temperature reactions under pressure. Consider the following when working with either apparatus:

  • Ensure that the heating element is completely enclosed. 
  • For air baths constructed of glass, wrap the vessel with heat resistant tape to contain the glass if it should break. 
  • Sand baths are generally preferable to air baths. 
  • For tube furnaces, carefully select glassware and metal tubes and joints to ensure they are able to withstand the pressure. 
  • Follow safe practices outlined for both electrical safety and pressure and vacuum systems. 

Heat Guns

Laboratory heat guns are constructed with a motor-driven fan that blows air over an electrically heated filament. They are frequently used to dry glassware or to heat the upper parts of a distillation apparatus during distillation of high-boiling materials.

Read the Heat Gun Advisory for more information on proper selection and use of a heat gun for research operations.

Microwave Ovens

Microwave ovens used in the laboratory may pose several different types of hazards.

  • As with most electrical apparatus, there is the risk of generating sparks that can ignite flammable vapors.
  • Metals placed inside the microwave oven may produce an arc that can ignite flammable materials.
  • Materials placed inside the oven may overheat and ignite.
  • Sealed containers, even if loosely sealed, can build pressure upon expansion during heating, creating a risk of container rupture.

To minimize the risk of these hazards, 

  • Never operate microwave ovens with doors open in order to avoid exposure to microwaves.
  • Do not place wires and other objects between the sealing surface and the door on the oven’s front face. The sealing surfaces must be kept absolutely clean.
  • Never use a microwave oven for both laboratory use and food preparation.
  • Electrically ground the microwave. If use of an extension cord is necessary, only a three-wire cord with a rating equal to or greater than that for the oven should be used.
  • Do not use metal containers and metal-containing objects (e.g., stir bars) in the microwave. They can cause arcing.
  • Do not heat sealed containers in the microwave oven. Even heating a container with a loosened cap or lid poses a significant risk since microwave ovens can heat material so quickly that the lid can seat upward against the threads and containers can explode.
  • Remove screw caps from containers being microwaved. If the sterility of the contents must be preserved, use cotton or foam plugs. Otherwise plug the container with kimwipes to reduce splash potential.

Ultrasonicators (top)

Human exposure to ultrasound with frequencies between 16 and 100 kilohertz (kHz) can be divided into three distinct categories: airborne conduction, direct contact through a liquid coupling medium, and direct contact with a vibrating solid.

Ultrasound through airborne conduction does not appear to pose a significant health hazard to humans. However, exposure to the associated high volumes of audible sound can produce a variety of effects, including fatigue, headaches, nausea and tinnitus. When ultrasonic equipment is operated in the laboratory, the apparatus must be enclosed in a 2-cm thick wooden box or in a box lined with acoustically absorbing foam or tiles to substantially reduce acoustic emissions (most of which are inaudible). 

Direct contact of the body with liquids or solids subjected to high-intensity ultrasound of the sort used to promote chemical reactions should be avoided. Under sonochemical conditions, cavitation is created in liquids, and it can induce high-energy chemistry in liquids and tissues. Cell death from membrane disruption can occur even at relatively low acoustic intensities. 

Exposure to ultrasonically vibrating solids, such as an acoustic horn, can lead to rapid frictional heating and potentially severe burns. 

Centrifuges (top)

Centrifuges should be properly installed and must be operated only by trained personnel. It is important that the load is balanced each time the centrifuge is used and that the lid be closed while the rotor is in motion. The disconnect switch must be working properly to shut off the equipment when the top is opened, and the manufacturer’s instructions for safe operating speeds must be followed.

For flammable and/or hazardous materials, the centrifuge should be under negative pressure to a suitable exhaust system. 

Rotary Evaporators (top)

Glass components of the rotary evaporator should be made of Pyrex or similar glass. Glass vessels should be completely enclosed in a shield to guard against flying glass should the components implode. Increase in rotation speed and application of vacuum to the flask whose solvent is to be evaporated should be gradual.

Autoclaves (top)

The use of an autoclave is a very effective way to decontaminate infectious waste. Autoclaves work by killing microbes with superheated steam. The following are recommended guidelines when using an autoclave: 

  • Do not put sharp or pointed contaminated objects into an autoclave bag. Place them in an appropriate rigid sharps disposal container.
  • Use caution when handling an infectious waste autoclave bag, in case sharp objects were inadvertently placed in the bag. Never lift a bag from the bottom to load it into the chamber. Handle the bag from the top. 
  • Do not overfill an autoclave bag. Steam and heat cannot penetrate as easily to the interior of a densely packed autoclave bag. Frequently the outer contents of the bag will be treated but the innermost part will be unaffected. 
  • Do not overload an autoclave. An overpacked autoclave chamber does not allow efficient steam distribution. Considerably longer sterilization times may be required to achieve decontamination if an autoclave is tightly packed. 
  • Conduct autoclave sterility testing on a regular basis using appropriate biological indicators (B. stearothermophilus spore strips) to monitor efficacy. Use indicator tape with each load to verify it has been autoclaved. 
  • Do not mix contaminated and clean items together during the same autoclave cycle. Clean items generally require shorter decontamination times (15-20 minutes) while a bag of infectious waste (24" x 36") typically requires 45 minutes to an hour to be effectively decontaminated throughout. 
  • Always wear personal protective equipment, including heat-resistant gloves, safety glasses and a lab coat when operating an autoclave. Use caution when opening the autoclave door. Allow superheated steam to exit before attempting to remove autoclave contents. 
  • Be on the alert when handling pressurized containers. Superheated liquids may spurt from closed containers. Never seal a liquid container with a cork or stopper. This could cause an explosion inside the autoclave. 
  • Agar plates will melt and the agar will become liquefied when autoclaved. Avoid contact with molten agar. Use a secondary tray to catch any potential leakage from an autoclave bag rather than allowing it to leak onto the floor of the autoclave chamber. 
  • If there is a spill inside the autoclave chamber, allow the unit to cool before attempting to clean up the spill. If glass breaks in the autoclave, use tongs, forceps or other mechanical means to recover fragments. Do not use bare or gloved hands to pick up broken glassware. 
  • Do not to leave an autoclave operating unattended for a long period of time. Always be sure someone is in the vicinity while an autoclave is cycling in case there is a problem. 
  • Autoclaves should be placed under preventive maintenance contracts to ensure they are operating properly. 

Electrophoresis Devices (top)

Precautions to prevent electric shock must be followed when conducting procedures involving electrophoresis. Lethal electric shock can result when operating at high voltages such as in DNA sequencing or low voltages such as in agarose gel electrophoresis (e.g., 100 volts at 25 milliamps).These general guidelines should be followed:

  • Turn the power off before connecting the electrical leads 
  • Connect one lead at a time, using one hand only 
  • Ensure that hands are dry while connecting leads 
  • Keep the apparatus away from sinks or other water sources 
  • Turn off power before opening lid or reaching inside chamber 
  • Do not override safety devices 
  • Do not run electrophoresis equipment unattended. 
  • If using acrylamide, purchase premixed solutions or pre-weighed quantities whenever possible 
  • If using ethidium bromide, have a hand-held UV light source available in the laboratory. Check working surfaces after each use. 
  • Mix all stock solutions in a chemical fume hood. 
  • Provide spill containment by mixing gels on a plastic tray 
  • Decontaminate surfaces with ethanol. Dispose of all cleanup materials as hazardous waste. 

Glassware (top)

Although glass vessels are frequently used in low-vacuum operations, evacuated glass vessels may collapse violently, either spontaneously from strain or from an accidental blow. Therefore, pressure and vacuum operations in glass vessels should be conducted behind adequate shielding. It is advisable to check for flaws such as star cracks, scratches and etching marks each time a vacuum apparatus is used. Only round-bottomed or thick-walled (e.g., Pyrex) evacuated reaction vessels specifically designed for operations at reduced pressure should be used. Repaired glassware is subject to thermal shock and should be avoided. Thin-walled, Erlenmeyer or round-bottomed flasks larger than 1 L should never be evacuated.

Vacuums (top)

Vacuum pumps are used in the lab to remove air and other vapors from a vessel or manifold. The most common usages are on rotary evaporators, drying manifolds, centrifugal concentrators (“speedvacs”), acrylamide gel dryers, freeze dryers, vacuum ovens, tissue culture filter flasks and aspirators, desiccators, filtration apparatus and filter/degassing apparatus.

The critical factors in vacuum pump selection are:

  • Application the pump will be used on
  • Nature of the sample (air, chemical, moisture)
  • Size of the sample(s)

When using a vacuum pump on a rotary evaporator, a dry ice alcohol slurry cold trap or a refrigerated trap is recommended. A Cold Trap should be used in line with the pump when high vapor loads from drying samples will occur. Consult manufacturer for specific situations. These recommendations are based on keeping evaporating flask on rotary evaporator at 400 C. Operating at a higher temperature allows the Dry Vacuum System to strip boiling point solvents with acceptable evaporation rates.

Vacuum pumps can pump vapors from air, water to toxic and corrosive materials like TFA and methylene chloride. Oil seal pumps are susceptible to excessive amounts of solvent, corrosive acids and bases and excessive water vapors. Pump oil can be contaminated quite rapidly by solvent vapors and mists. Condensed solvents will thin the oil and diminish its lubricating poroperties, possibly seizing the pump motor. Corrosives can create sludge by breaking down the oil and cause overheating. Excess water can coagulate the oil and promotes corrosion within the pump. Proper trapping (cold trap, acid trap) and routine oil changes greatly extend the life of an oil seal vacuum. Pump oil should be changed when it begins to turn a dark brown color.

Diaphragm pumps are virtually impervious to attack from laboratory chemical vapors. They are susceptible to physical wearing of the membrane if excessive chemical vapors are allowed to condense and crystallize in the pumping chambers. A five minute air purge either as part of the procedure or at day’s end will drive off condensed water vapors and further prolong pump life.

Hazardous chemicals can escape from the vacuum pump and pump should be place in the hood. Cold traps and acid traps can be helpful, but if allowed to thaw or saturate, they can lose their effectiveness.

Section 7J: Particularly Hazardous Substances
Section 7H: Pressure and Vacuum Systems

       
       
     

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