Safe Laboratory Practices
and Procedures
Chemistry Department
University of Cincinnati
Outline
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Emergency Procedures
Required Procedures
Safe Laboratory Practices
Chemical Hazard and Risk Information
Gas Cylinders
Horror Stories
Future Safety Plans
Emergency Response Plan
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All personnel must begin evacuation at once. Pull the manual
fire alarm, if building is not already in alarm to alert all
occupants. Call 9-1-1 once outside the building.
Take any valuable personal items.
As you leave the laboratory or office, quickly check to determine
if all co-workers have begun to evacuate. The last person out
should close, but not lock the door to the laboratory.
All personnel must familiarize themselves with the most rapid
means of escape from their laboratory or office. Evacuation
maps are posted in hallways near the elevators.
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An alternative means of escape should also be noted for those
emergencies where the first exit is blocked.
Never use the elevators if a fire alarm activates; take the stairs
instead.
Once you have evacuated the building, assemble at designated
areas to enable safety monitors to ensure everybody has
evacuated.
Assemblage areas are outside Crosley Tower on the second
floor, outside Reveischl Hall on the fourth floor or the sixth floor.
Designated Safety Monitors (faculty) will ensure their assigned
areas are evacuated.
You cannot enter the building until given permission by
Emergency Response Personnel.
A. Fire Emergency: Controllable Fire
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Pull the manual fire alarm, if building is not already in alarm to
alert all occupants. Call 9-1-1 once outside the building.
Notify everyone in the immediate area of the danger.
Non-essential personnel should evacuate the area.
You may fight the fire yourself if:
a) You have been trained in the use of fire extinguishers,
b) the fire is small (size of a wastebasket) and confined to a
specific area (such as a fume hood),
c) you are reasonably certain that the fumes that are generated
are not toxic
d) you have a clear means of escape if the fire proves to be out
of your control.
Never turn your back on a fire even if you believe it to be
extinguished.
Report the fire to the floor’s Safety Monitor (faculty) and UC Fire
Prevention (556-4992) even if extinguished.
A. Fire Emergency: Uncontrollable Fire
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Notify everyone in the immediate area of the danger.
Pull the manual fire alarm, if building is not already in alarm to
alert all occupants.Call 9-1-1 once outside the building.
Evacuate the area
Close doors and windows if possible
Evacuate the building and go to an assembly area.
B. Persons with Disabilities (Fire)
Public Safety guidelines permit individuals with disabilities the
option of staying in the building during an emergency situation if
they are non-ambulatory, or where elevator assistance is essential
for their evacuation, and they are located either above or below the
ground floor. A non disabled person should stay with the disabled
to assist if necessary. Detailed instructions are available by visiting
University of Cincinnati Fire Prevention’s website at
http://www.uc.edu/techserv/fireprev.htm. The individual, or a person
designated to be with them should:
• If on ground floor, exit the building as usual.
• If above, or below ground, seek a safe place- preferably a
designated area of refuge (none are currently located in Logan
Hall), or a room with an exterior window, a telephone, and a solid
door.
• Telephone Emergency Dispatch (9-1-1) and advise them you are
a person with a physical disability. Give your exact location –
Building, Floor and Room number and the number of the phone
number you are calling from.
• The Fire Department will determine if evacuation of these
occupants is necessary.
C. Chemical Release: Controllable
Chemical Release without Injury or Illness
• Stop the release of chemicals if possible and safe to do so.
• For flammable material, disconnect all ignition sources.
• Notify all personnel in the immediate area of the danger.
• Evacuate all non-essential personnel.
• Once the chemicals are contained, perform appropriate clean-up
procedures.
• Clean-up procedures should follow instructions on appropriate kits
and in accordance with Material Safety Data Sheets.
a) Appropriate protective gear needs to be worn.
b) For flammable liquids, Flammable solvents may be absorbed with
dry absorbents, such as commercially available universal spill control
products, vermiculite or sand. Use spark-free utensils to pick-up the
waste.
c) For toxic liquids, spill control and clean-up, various dry absorbents
may be used. These include commercially available universal spill
control products, vermiculite and sand. Waste material must be
bagged and tagged for disposal. Store waste in a ventilated area.
d) For corrosive liquids:
1) Small acid spills can be neutralized by applying sodium
bicarbonate or a similar weak base, and allowing reaction to
occur. The neutralized material can then be swept up and
disposed via regular waste disposal or through the sewage
system, if the quantity is not large.
2) Small caustic spills can be neutralized by the addition of citric
acid, or other similar weak acid. The reacted material can then
be swept up and disposed of via regular waste collection or the
sewage system, if the quantities are not large.
3) Dry absorbents may also be used on either acidic or basic spills.
Materials suitable for use as absorbents include commercially
available spill control products, vermiculite, and sand.
4) Hydrofluoric acid spills require special spill control products. Do
not use sand or vermiculite. Follow specific guidelines in the
Material Safety Data Sheet.
e) For solid chemicals:
1) Nontoxic solids can be swept up (avoiding breathing dust) and
disposed of in a waste container.
2) Oxidizers must not contact paper, e.g., trash cans.
3) Toxic or reactive solids must be handled carefully and in
accordance with instructions given in Safety Data Sheets.
4) Small mercury spills can be collected using a vacuum device or
gathered using an amalgamating powder, e.g., sulfur. The
waste needs to be tagged for disposal. Large mercury spills
require a Mercury Vacuum Cleaner.
f) The waste material needs to be disposed of properly.
C. Chemical Release: Uncontrollable
Chemical Release without Injury or Illness
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Get away from the area until proper provisions can be made.
Notify personnel in the immediate area of the chemical release.
Pull the manual fire alarm, if building is not already in alarm to
alert all occupants.Call 9-1-1 once outside the building.
If the chemical spill is large, in a poorly ventilated area or is
extremely toxic (that is, if it is beyond the capabilities of the
attending personnel or of the equipment at hand), call 9-1-1 for
the HazMat Team for assistance.
Close the door and windows, if possible.
Report the incidence to a Safety Monitor
C. Chemical Release: with Injury or
Illness
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Follow the Chemical Release Plan.
If you are already in the exposure area, remove the injured
person from the area of exposure without injuring yourself.
If you are not initially in the exposure area, do not attempt to
remove the victim or enter the exposure area. You may become
a victim yourself.
If the situation is life threatening, call 911 immediately.
Begin decontamination by moving the victim under a safety
shower or over an eyewash station. Flush contaminated areas
thoroughly.
If possible, determine which chemical was released.
Monitor the victim for an extended period of time especially prior
to the arrival of medical personnel. Symptoms may take a while
to appear.
All injured personnel need to be examined by a doctor before
returning to work.
D. In Case of Bomb Threats or
Suspicious Activities
When you receive a bomb threat or observe a suspicious person
or activity:
• Notify UC police. Do not use a cell phone to report bomb threats.
• Do not attempt to remove or open suspicious packages
• Suspicious individuals can be approached; however, do not
confront them in a threatening manner.
E. Severe Weather
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If severe weather is detected (severe thunderstorms, tornadoes
etc) the campus severe weather warning system will sound,
instructing all occupants to:
Evacuate your area using either stairwell. Elevators may only be
used by individuals with disabilities.
Severe weather safety locations for Crosley Tower are noted as
the basement corridor. The public restrooms may also be used
as a shelter.
Await further instruction from Public Safety or Cincinnati Fire
Department before leaving the assembly area.
Required Procedures
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YOU MUST:
Wear appropriate eye covering
Wear proper clothing (cotton lab coat and secure shoes)
Not eat or drink in the lab or apply cosmetics, and no food or
beverage containers in the lab, empty or otherwise
Always treat reagents as if they are poisonous, toxic, and
dangerous
Keep reagent containers sealed and clean (put the receiving
date on the container)
Understand the reactivity, toxicity, and hazards of the reagent
that you are using (check its MSDS sheet)
Dispose of materials properly and in a timely manner
Keep the lab clean and uncluttered
Not transfer chemicals in a public elevator; use the dumb waiter
Not become complacent
Safe Laboratory Practices
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All procedures involving harmful chemicals must be done in a
fume hood whenever possible. Check the hood with a kimwipe
before using it to make sure it is pulling air.
The sash of the fume hood should be positioned as low as
possible while maintaining secure handling of the materials.
Appropriate gloves should be worn when handling chemicals.
Open all chemical containers and the shipping boxes in a
ventilated hood with sash lowered and the container pointed
away from your face.
When removing a septum or stopper from your flask, first open
the side that is away from your face (it may be under pressure).
Only heat solutions in open systems (drying tube, Ar
environment etc.) unless the container is designed to handle
high pressures and a blast shield is used.
No open flames or ignition sources near volatile, flammable
materials (acetone, ether, etc.).
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Do not use chipped or cracked glassware – dispose of it.
Do not force anything e.g. when opening glassware, pushing
glassware through stoppers, removing valves from cylinders.
You can lubricate glass joints.
Use appropriately sized syringes to transfer reagents and
solvents.
Use a cannula for large volumes.
Syringe locking is likely for moisture sensitive materials.
Ask for help or guidance when doing a new procedure, using a
new reagent, or before you start forcing something.
Blast shields should be used when manipulating explosive
materials outside the fume hood. Rotoevaporation is a
dangerous time for chemicals, which are often being dried and
heated. Use a high capacity vacuum pump if a solvent has a
high boiling point.
Monitor distillations; do not heat to dryness; do not have the hot
plate set to a temperature significantly higher than the boiling
point of the solvent.
Add reagents to solvent.
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Make sure weigh papers that contained reagents are quenched
prior to disposal.
Quench all reactive material prior to disposal (e.g., use tbutanol)
Do not mix reactive materials in waste cans (e.g. no strong
oxidizers with waste materials).
When finished with liquid nitrogen traps: remove trap from liquid
nitrogen, open the system, turn off pump (liquid O2—boom!).
Make sure you consider heats of mixing when quenching
reactions or doing extractions.
Carefully vent separation funnels, pointing them away from your
face.
Transfer materials safely (inside and outside the lab). Do not
wear gloves in the hallways or elevators, in the main or business
office, and so forth. Use the dumbwaiter to transfer harmful
materials between floors.
Label containers holding prepared solutions - contents, hazards
and concentration.
Do not transport or store NMR tubes in your pockets
Hazard and Risk Information
Internet Resources
MSDS Directory (http://www.ilpi.com/msds/index.html)
National Institute for Occupational Safety and Health (NIOSH)
Pocket Guide to Chemical Hazards and International Chemical
Safety Cards (http://www.cdc.gov/niosh/npg/npg.html)
SAFETY Listserv Archives (http://hazard.com)
Sigma Aldrich (http://www.sigmaaldrich.com/united-states.html)
MSDS sheets
From NIOSH Pocket Guide
Gas Cylinders
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Compressed gas cylinders have an enormous amount of stored
energy just waiting to be released (2200 lbs. p.s.i. stored
pressure)
Can be released if they are dropped, mishandled or abused.
If the regulator or valve connection is broken, exhaust vapor
under pressure creates a force that moves the cylinder like a jet
engine
Damage to property and injury or death to people could easily
result when this type of energy is released, not to mention the
gas is also released.
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Handling Cylinders
Assure all cylinders are properly restrained from tipping and falling
against objects. Never have more than 5 cylinders restrained
together in one place. Pedestal base, chain or belt restraints all work
well. A proper restraint is critical.
Don’t move a cylinder with a regulator attached; have its valve cap
in place when moving it.
Never transport cylinders horizontally. Use a cylinder cart with a
restraint belt or chain and keep them nearly upright.
Assure the contents of all your cylinders are properly labeled. Know
the hazards of the contents by reviewing the Material Safety Data
Sheet (MSDS) for each gas you use.
Remember, many gases can cause frostbite upon contact, and may
not be smelled or seen.
If you hear vapor escaping, ensure the valve is closed. If you still
hear vapor, you may have a leak. A soap bubble solution can help
you find a small leak.
Never use grease or oil near oxygen cylinder regulators or valves.
Spontaneous ignition could occur.
Horror Stories
Accident:
Transferring ~ 50 mL t-butyl lithium (in pentane) with a 50 mL
syringe
Barrel came out of the syringe
Flash fire ignited her clothes (sweater)
She ran the wrong direction from the safety shower
She eventually died from the accident
Better Lab Practice:
Use a syringe with a greater capacity than the liquid to be
transferred
Better to transfer large volumes via a cannula (double headed
needle)
Operate the syringe in the fume hood with sash as low as possible
(arms exposed only) and pull in a direction away from the body
Wear a cotton lab coat, which is less likely to catch fire (never wear
a lab coat made of synthetics – they melt and burn while stuck
to your skin)
Know where the safety shower, fire blanket and eye wash are
Accident:
Synthesis of an energetic material, most likely nickel hydrazine
perchlorate
Not supposed to make more than 100 mg of the material, the
student synthesized 10 g.
Because the product was lumpy, student placed his portion into a
mortar. He believed that the compound was safe when “wet,” so
he added some hexane and—wearing safety goggles but
working at a bench in the middle of the lab, with no blast
shield—“very gently, very, very gently” used a pestle to try to
break up the chunks,
He set down the mortar and took off his goggles. Then he decided
to give the compound one last stir. The mortar exploded in his
hands. He “lost three digits on his left hand, severely lacerated
his right hand,
perforated his left eye, scratched his right eye and had superficial
cuts to the parts of his body that were exposed
The student’s laboratory notebook provides no detail about what
exactly the two were doing in the lab on the day of the incident.
During that time, the notebook includes seminar notes, other
notes seemingly unrelated to his research, and only vague
descriptions of experimental work: One apparent synthesis is
documented in an undated entry simply as “Ni(NO3)2 +
hydrazine / 10 g / purple ppt forms immediately / also an
exothermic rxn.” The other student’s “notebook” is a typed
summary of reaction protocols and observations, also without
dates.
Interviews with other researchers who worked in the lab indicate
that his labmates were disturbed by his conduct in the lab prior
to the incident. His space was disorganized, items were not
labeled, and “there had been conflicts over work space,
cleanliness of the lab and use of chemicals,”
Student started scaling up syntheses, first to 1 to 3 g and then to 5
g. The researcher told the student the scale-up was
inappropriate; student reportedly responded that things were
“just fine.” The researcher apparently did not report the scale-up
Student transported as much as “several grams of compounds” at a
time in glass vials in a backpack or coat pocket, a researcher
who helped the student told EH&S investigators. He “was told
that a metal container would be better for the transport, but he
continued to bring them in a glass vial,”
Several vials of material were found in the student’s home. The
containers were largely unlabeled, except for one marked
“TATP,” most likely for triacetone triperoxide. The bomb squad
destroyed the items. Student brought vials home from the lab
because he absent-mindedly left them in his pockets
Other powders and liquids found in the lab could not be identified
because of improper labeling.
Better Lab Practice:
Wear appropriate eye protection
Use blast shields when working with energetic materials
The first rule of handling energetic materials is to keep quantities
small
Do not become complacent because accidents haven’t happened –
yet...
Let advisor know before scaling up materials
Follow safety policies to the letter
Transport materials in appropriate containers
Write down and follow safe protocols
Accident:
Rotoevaporation of a peroxide
Student removed flask from bump trap and the flask disappeared in
an explosion.
Student was not hurt, luckily the glass returned to sand
Rotoevaporation of an azide
Gram scale reaction was evaporated to dryness and left heating for
a long time.
The flask exploded
Student suffered cuts on his face, chest, and hands
Better Lab Practice:
Use a blast shield when using a rotoevaporator with energetic
materials
Do not heat the solid excessively
If possible, just reduce the volume without reaching dryness
If possible, manipulate small scales of energetic/reactive materials
Future Safety Plans
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Research Group Chemical Hygiene Plan – describes safe
practices and procedures appropriate for that group.
Appendices – written by group members for particularly
harmful materials or procedures. These will be available for all
departmental members.
Self Safety Audits – Each group checks for safety violations.
Departmental Safety Audits – Members of the Safety Committee
will check for violations
Adding safety questions to research presentations
CHP Appendix Example
Piranha Solution
Standard Operating Procedure
Neil Ayres Research Group
Overview:
Piranha solution is a powerful oxidizing solution used to remove
organic residues from substrates. In this lab, we typically use a
7:3 mixture of concentrated sulfuric acid and 30% hydrogen
peroxide to etch silicon wafers. Piranha solutions are extremely
energetic and may result in explosion or skin burns if not
handled with extreme caution. Any researcher handling piranha
should wear a laboratory coat, closed toe shoes, long pants,
safety glasses, and neoprene or polyvinyl chloride gloves (nitrile
gloves do not supply sufficient protection). Piranha solution must
be prepared under the hood with the glass shield pulled down
as far as it can go within a comfortable working distance. The
solution should be prepared in a glass container-never in plastic.
NEVER allow any organic to come in contact with piranha (this
will cause an explosion).
Always add the acid SLOWLY to the hydrogen peroxide. After the
solution is prepared, begin heating the solution to ~ 70 oC. This
should be done SLOWLY in 5-10 degree increments to avoid
generating an extreme amount of heat (this reaction is exothermic).
Monitor the solution as it is heating, and only increase the
temperature when the solution is calm. Always use forceps to insert
and remove wafers from the solution. NEVER use your hands.
Emergency Procedures:
In case of skin contact: May cause skin burns. Remove
contaminated clothing and flush the skin with copious amounts of
water for at least 15 minutes. Seek medical attention.
In case of eye contact: Piranha is corrosive and irritating to the
eyes. Flush contaminated eye(s) immediately with copious
quantities of water for at least 15 minutes. Seek medical attention
immediately.
In case of inhalation: May irritate the respiratory tract. Conscious
persons should be assisted to an area with fresh, uncontaminated
air. Seek medical attention in the event of respiratory irritation,
cough, or tightness in the chest. Symptoms may be delayed.
In case of ingestion: Not a likely route of exposure.
Storage:
Do not store piranha. Mix fresh solution for each use.
Disposal:
Allow the solution to cool completely before removing it from the
hood. Turn on the water in the sink and, using forceps, carefully
transport the solution to the sink. With the forceps, dump the
solution, and leave the dish to flush under the running water for
several minutes before removing.
Summary
• You are responsible for laboratory safety
• Always treat reagents as if they are poisonous, toxic,
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and dangerous
Understand the reactivity, toxicity, and hazards of the
reagent that you are using (check its MSDS sheet)
Ask for help or guidance when doing a new
procedure or working with new materials
You must not become complacent