Major Hazards in Laboratory
Manish Kumar (Ph.D)
Laboratory Guidance and Safety Lecture
Potential hazards in laboratory
Potential Hazards Type of Hazards
High Pressure Gas Explosion, deoxygenation, poisoning etc
Autoclave High temperature and High Pressure
Centrifuge machine Rotor, shaft breakage and Explosion
High-temperature furnace High temperature (~6000C or higher)
ICP/OES, ICP/MS High temperature
Microorganisms Infection and Biohazard
Chemicals Poisoning, carcinogenic exposure, cause
of fire
Electric lines Electric shock and electric fire
Glass equipments Injury
Syringes Injury and microbial infection
Tools Injury 2
Dangerous Equipments
Autoclave
High Pressure
gas
High Temperature
Oven Be careful when you use these apparatus
Few others
Natural Hazard/Earthquake
• Avoid putting heavy thing at
high place.
• Avoid making complicated
electric lining.
• Simulate what to do in case
of emergency.
• Organize regular drill for
specific natural hazard.
Accidents/First Aid
• If a person is injured: Do not
move the person unless there is
a danger to life.
• Switch off any electricity or
mechanical equipment (if it is
safe to do so).
• Call for a First Aid
Officer/hospital extension
number.
• Keep a First Aid kit around the
buildings and in laboratories. 4
Hazards specific to High Pressure Cylinders
• Rocketing: Very dangerous if the high pressure inside (150atm)
is released. Fix a cylinder to its holder with an appropriate
regulator.
• Explosion: Be very careful when u handle a flammable gas like
Hydrogen. Leak of H2 gas can cause explosion. It is used for
GC/FID (flame ionization detector for gas chromatography).
You need to check a) if hydrogen flame is surely on, and b) if
there is enough air in air cylinder to support the flame.
• Deoxygenation: A sudden leak of gas (N2) can decrease the O2
concentration in air and if you inhale such air, you might faint
and fail to escape.
• Falling: Cylinders are pretty heavy. You may fall down with
cylinder while moving it.
5
Hazards aspects about chemicals
• Explosiveness & Flamableness:
– Avoid applying physical shock, fire, heat...
– Avoid putting close to electricity, spark may cause fire
– Notify people around about the risk. Put notice.
• Toxicity:
– Avoid Exposure
– Use gloves, eye protect,..
– Handle evaporative chemicals in a fume hood.
Source of safety information:
1) International Chemical Safety Cards (ICSCs)
http://www.cdc.gov/niosh/ipcs/icstart.html( )
• http://www.ilo.org/public/english/protection/safework/cis/pr
oducts/icsc/index.htm
6
Handling aspects of autoclave
• What to check before use:
– Water level in the autoclave.
– Water level in the drain
– Exhaust valve
– (must be closed)
• What to avoid when the autoclave is in use:
– Never open the cover
– Even if the internal pressure turns to zero, it is recommended to wait a
little bit more.
• What to do after use:
– In case glass were eloded in an autoclave, remove the glass particle and
change the water.
• How to maintain: Periodically replace water inside autoclave.
7
Microbial Laboratory related Hazards
Pathogens and toxins considered to have the potential to
pose a severe threat to human, animal, or plant health and
safety.
• Viruses
• Bacteria
• Fungi
• Toxins
8
Biohazard:
An agent of biological origin that has
the capacity to produce harmful effects
on humans; i.e. microorganisms, toxins
and allergens derived from those
organisms, and allergens and toxins
derived from plants or animals.
Definition
9
Biosafety:
Applying a combination of laboratory practices and procedures, laboratory facilities, and safety equipment when working with potentially infectious microorganisms.
10
Risk Assessment:
Addressing laboratory activities involving infectious or potentially infectious material and
implementing measures to reduce the worker’s and environment’s risk of exposure to an agent to an
absolute minimum.
11
Biosecurity:
Protection of high-
consequence microbial agents
and toxins, or critical relevant
information, against theft or
diversion by those who intend
to pursue intentional misuse.
12
• Biosecurity refers to ensuring the security of biological materials to prevent theft, illicit use, or release
• Biosafety focuses on reducing exposure to and release of biological materials
• Both involve conducting a risk assessment to mitigate risks
Biosecurity vs. Biosafety
13
Keep the entrance door of microbiological
laboratory always locked.
Infectious viruses are supposed to be stored in
sealed-up containers. When opening the seal,
everything has to be done in the safety cabinet.
When using infectious viruses, post a notice on the
door of the laboratory .
Keep laboratory always clean and neat to avoid
contamination and leakage of infectious microbes.
14
15
Notification of Infectious Microbes
• When one obtain a new infectious
microbe, one have to notify the Genetic
Experiment Management Committee of
the use and storing of the microbe.
• If one plan to get new infectious
microbe, their SUPERVISOR should be
consulted.
16
Waste Management
• All microbiological wastes must be
autoclaved first.
• Use special bags to discard
autoclaved with residuals.
• Latex gloves must be stored and
thrown away as pseudo infectious
wastes. 17
General Cautions • Close the windows and door during experiments.
• Do not take away laboratory equipment’s out of
the lab.
• Disinfect the safety cabinet(by ethanol and UV
lamp) when experiment is over.
• Use micropipette.
• Wash hands before leaving laboratories.
• Try to minimize the occurrence of aerosol
contamination.
18
Contd.
• Prepare and use of lab coat in
microbiological lab.
• Use equipped sandal when getting into
microbiological lab.
• Do not enter the room without
accompanying a well-trained student if you
are a beginner.
• Limit the refrigerators, freezers and
autoclave machines for clean use.
19
Secure escape route
Do not put anything in front of the windows with the red triangle mark.
Fire fighters will use them in case of accidents.
Do not put anything on the experimental shelf. Take careful protection
for earthquake.
Do not put unnecessary goods in the halls/corridors.
Be careful when you use emergency stairs because they are slippery.
GENERAL CAUTIONS FOR
CHEMICALS
1. Never return chemicals back to the bottle if taken out once.
2. Use a dry and clean spoon to handle solid chemicals.
3. When you take liquid chemicals, pour some to a small beaker, then handle with a pipette. Do not take liquid directly from the original container by using a pipette.
4. Use up an old chemical bottle, then open a new on, in case when there is stock of the chemical.
5. Phenol is solid at room temperature. Soak it in hot water bath before handling it.
21
Contd. 6. When you make diluted sulfuric acid solution, pour conc.
Sulfuric acid into water. (Never do the opposite, because it is very dangerous.)
7. When you have used conc. Sulfuric bottle, please wipe off the remaining sulfuric acid around the mouth of the bottle. The remaining drop will reach bottom of the bottle, and contaminates the chemicals drawers, and also, surface of benches. Finally, it could cause holes on your cloths or do harm on your skin.
8. The reagents you have prepared should be replaced into a reagent bottle and then stored. Do not store them in measuring flasks.
9. Information should be recorded on the reagents` labels including: reagent name, concentration, date of preparation, your name.
22
To avoid production of unknown
chemicals
HCl (dil.)
0.01 N
3/2/12
Jyoti Reagent Name
Concentration
Date
User Name Good example Bad example
The skin: Neutralize with 0.1
M NH4OH or 1% NaHCO3
and then wash with water.
The eyes and mouth:
Neutralize with 0.1% NaHCO3
and then wash with water.
Clothes: Neutralize with 0.1M
NH4OH and then wash with
water
0.1%
NaHCO3
0.1 M
NH4OH
1
2
Immediately wash with plenty of
water when strong acid adheres on
the skin or clothes.
First aid - acid burn
First aid- alkali burn
Wash with plenty of water in the same way as acid burn.
The skin: Neutralize with 0.1%
CH3COOH (acetic acid) and then
wash with water.
The eyes and mouth: Neutralize
with ca.2% H3BO3 (boric acid)
and then wash with water.
Clothes: Neutralize with 0.1%
CH3COOH and then wash with
water.
2
1
3 2
0.1%
CH3COOH
2% H3BO3
BASIC METHOD TO WASH
GLASSWARE 1. Working detergent solution for washing: Prepare a detergent solution
by diluting a concentrated detergent several hundred times. ( For
example, add around 2 to 3ml of detergent to 1L of tap water.)
2. Wash inside and outside of the glassware by using a brush or a piece
of sponge. Wash not only inside, but also outside. Do not use black
brushes to wash measuring flasks or measuring cylinders because they
may scratch internal glass wall. Use white brush instead. (“white is
soft, black is hard”)
3. Rinse with tap water to remove remaining detergent.
4. Rinse with deionized water or RO (reverse osmosis) water three times.
(No need to use expensive ultrapure water.)
5. Dry glassware in drying shelves. If drying shelves are full, try to move
dried glassware to proper spaces. (Separate the space for drying and
storage.)
26
Common Vs Private
1.Return common tools/stuffs to their original place
Beakers, measuring cylinders, tools, spoons, micropipettes, chemicals….
2. Please keep always clean around common spaces
Keep clean around balances.
Clean up workplaces, especially after using a drill or saw.
Remove your stuffs after using common analytical tools.
Always try to keep our laboratory clean and comfortable.
27
Contd.
3. Wash your stuffs without delay
Wash stuffs as soon as possible. Never leave them in a sink, which makes others in trouble.
4. Mark your belongings
Put your name clearly on your samples, reagents, and so on.
Do not use stuffs such as reagents, tools and wash bottles on other`s bench without permission.
5. Pay attention to the stock of common goods
When consumables for common use are running out, order the new one as a stock.
28
WASHING BOTTLES IN
DIFFERENT COLOURS
Washing bottles are indicated with color, as shown on the right.
Ultra Pure Water RO/deionized water Tap water
(Milli-Q)
There are also special wash bottles filled with organic solvents such as ethanol or
acetone.
Download me from the following address:
http://www.tezu.ernet.in/denvsc/
29
First aid- external injury
Arrest of bleeding, prevent from infection and affliction removal
Normal injury
Wash and make the stain go
away.
Astriction (pressure point) with
sterile gauze.
Disinfection and bandage if
needed.
Cool down is effective for
turgescence and aching.