Major Hazards in Laboratory

Manish Kumar (Ph.D)

Laboratory Guidance and Safety Lecture

manishk@tezu.ernet.in 1

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.

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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

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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.

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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

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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

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Biosafety:

Applying a combination of laboratory practices and procedures, laboratory facilities, and safety equipment when working with potentially infectious microorganisms.

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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.

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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.

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• 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

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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.

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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.

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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.

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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.

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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.

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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.

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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.)

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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.

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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.

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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/

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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.