Major Hazards in Laboratory
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1 Major Hazards in Laboratory Manish Kumar (Ph.D) Laboratory Guidance and Safety Lecture 1
2 Potential hazards in laboratory Potential Hazards High Pressure Gas Autoclave Centrifuge machine High-temperature furnace ICP/OES, ICP/MS Microorganisms Chemicals Electric lines Glass equipments Syringes Type of Hazards Explosion, deoxygenation, poisoning etc High temperature and High Pressure Rotor, shaft breakage and Explosion High temperature (~600 0 C or higher) High temperature Infection and Biohazard Poisoning, carcinogenic exposure, cause of fire Electric shock and electric fire Injury Injury and microbial infection Tools Injury 2
3 Dangerous Equipments High Pressure gas Autoclave Be careful when you use these apparatus High Temperature Oven
4 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
5 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 H 2 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 (N 2 ) can decrease the O 2 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
6 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) ) oducts/icsc/index.htm 6
7 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
8 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
9 Definition 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. 9
10 Biosafety: Applying a combination of laboratory practices and procedures, laboratory facilities, and safety equipment when working with potentially infectious microorganisms. 10
11 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
12 Biosecurity: Protection of highconsequence microbial agents and toxins, or critical relevant information, against theft or diversion by those who intend to pursue intentional misuse. 12
13 Biosecurity vs. Biosafety 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 13
14 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 15
16 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
17 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
18 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
19 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
20 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.
21 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
22 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
23 To avoid production of unknown chemicals Bad example Reagent Name Concentration Date User Name HCl (dil.) 0.01 N 3/2/12 Jyoti Good example
24 First aid - acid burn Immediately wash with plenty of water when strong acid adheres on the skin or clothes % NaHCO 3 The skin: Neutralize with 0.1 M NH 4 OH or 1% NaHCO 3 and then wash with water. The eyes and mouth: Neutralize with 0.1% NaHCO 3 and then wash with water. Clothes: Neutralize with 0.1M NH 4 OH and then wash with water 0.1 M NH4OH 2
25 First aid- alkali burn Wash with plenty of water in the same way as acid burn. 1 The skin: Neutralize with 0.1% CH 3 COOH (acetic acid) and then wash with water. The eyes and mouth: Neutralize with ca.2% H 3 BO 3 (boric acid) and then wash with water. Clothes: Neutralize with 0.1% CH 3 COOH and then wash with water. 0.1% CH 3 COOH 3 2 2% H 3 BO 3
26 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
27 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
28 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
29 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: 29
30 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.
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