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CENTRAL UNIVERSITY OF RAJASTHAN Central University of Rajasthan

Laboratory Safety Training Dr. Janmejay Pandey

CENTRAL UNIVERSITY OF RAJASTHAN Central University of Rajasthan

Outline of the presentation:


1. 2. 3. 4. 5. 6. 7. Why is it important Some fundamental Concepts Safety in Chemical & Biological Labs Safety Supplies and Equipments Safe Laboratory Practices Safety from fire Your contribution towards making labs safe

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Why is it important ?

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C. U. Rajasthan- Motto

Education for sustainable Development Laboratory Safety !


A critical component of sustainable development

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Consequences of lack of lab safety measures !


1. Damage or loss of institutional property/ valuables. 2. Loss of important work/ results. 3. Most importantly .. Serious personal injuries, illness and

disabilities.

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Negligence of lab safety : A horror story

Sheri Sangji

1. 2.

UCLA (Dec. 18, 2008). A chemistry researcher working with a pyrophoric chemical got caught in a Laboratory fire. She suffered deep 3rd degree burns over 40 % of her body & 18 days later she died from this accident.
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CENTRAL UNIVERSITY OF RAJASTHAN Lack of lab safety training : Another tragic example
1. Georgia Institute of Technology (2003). An Afro- American Lab Technician attempted to bring down a non- labeled reagent bottle from a high shelf. The bottle slipped out of her hand, content of the bottle spilled over her. It turned out to be concentrated acid.

2.

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Do we need to be in accidents to learn lessons !

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Most frequently reported infections in Laboratory workers during last 10 years
Disease or Agent Mycobacterium tuberculosis Q fever Hantavirus Hepatitis B virus Brucella sp. Salmonella sp. Shigella sp. Hepatitis non-A, non-B No. of Cases 223 176 169 84 81 66 56 28

Cryptosporidium sp.
Total
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27 1074
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Flying laboratory instruments/ objects

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Injuries to Research Students/ Employee

Source: Howard Hughes Medical Institute, Office of Laboratory Safety


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Injuries to Laboratory Support Staff/ Random Visitor

Source: Howard Hughes Medical Institute, Office of Laboratory Safety


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Government Regulations for Lab Safety


Training is an important government regulation in western world
1. Regulatory requirement in several developed/ developing countries and organizations e.g. (NIH, CDC, OSHA, EPAUSA), (CSIRO- Aus), (EMBL- EU). 2. Institutional and Laboratory quality assessment & audit.

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Indian Status

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We need to learn the risks associated with labs in a holistic manner & also how to mitigate their hazardous effects. How can we make our labs safe !

Lab Safety Measures


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Key Concepts of Laboratory Safety


Laboratory safety revolves around

understanding, safe storage, usage & disposal of


hazardous components of lab work

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Key Concepts of Laboratory Safety


1. Understand 2. Organize 3. Ensure containment

4. Protect
5. Be prepared

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1. Understand
Prior preparation is extremely useful in ensuring lab safety. Physical: Chemical:
Get training for safe use of instrument. Use MSDS and other source about chemicals.

Biological:
Specific:

Read about hazardous effects of biological material.


Get necessary specific training e.g. radiological/ laser etc.

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2. Organize
Organize your work and work place before you start. Work diligently. Be alert and attentive. Dont work in lab in haste.

Ensure everything is safe before leaving the lab.

(Most laboratory accidents happen when people are distracted or hurried) Clinical Microbiol. Rev. (1995). 8: 389- 405
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3. Ensure containment
Keep hazardous material (chemicals, biological and physical hazards) within at least 2 levels of containments.
Contain any undesired spillage/ release of any and all hazardous material.

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4. Protect (yourself and co-workers)


Use appropriate protective and safety equipments.
1. Protective clothing (Lab cloth, Lab glasses, Gloves, Rubber Boots etc.).

2.
3. 4.

Chemical Fume Hood.


Laminar Air Flow and Bio-Safety Cabinets. Protective Working Shield.

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5. Be prepared (for emergencies)


1. Despite all the laboratory safety trainings, you may still encounter laboratory accidents

2. We must be adequately trained and prepared for such events.

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Safety in Chemical & Biological Labs

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Chemical hazards in a laboratory

Several forms of chemical hazards


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Explosive

Any chemical (solid, liquid or gas) that is in its unstable equilibrium. When initiated by shock they undergo rapid chemical change Have the tendency to change into more stable compound (usually gas) In the process of change they give out energy, heat, light & sound

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Characteristics of explosive
Can expand upto 10,000 15,000 times of their original volume Expansion may occur at a rate of 6,000 meters/ second Exert pressure of 110 metric ton/ square centimeter Cause temperature of 3,000- 4,000 oC within 10 seconds
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Corrosive

Corrosive chemicals (Acids, Bases, Strong Oxidizing Agent) Corrosive chemicals react with animal tissues & metals Cause burn and permanent tissue damage at the point of contact
Corrosives

can cause irreversible damage to eyes


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Corrosive Protection

Work in Chemical Fume Hood, Protect Eye, Face, Skin PVC and nitrile gloves often provide appropriate protection

Use splash goggles, face shield if splashing is possible


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(A- W of dilution) :
Add Acid gently to Water

1. Make sure you add a very small amount of the acid to the water. 2. Never add the water to a concentrated acid solution.

3. The Acid- Water mixing is an exothermic reaction and gives off loads of heat.
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Oxidizer

Oxidizers are not of themselves combustible, but can initiate and support combustion These are chemicals capable of supporting a fire by giving oxygen to the fire. Examples: Organic Peroxide with basic chemical structure R-O-O-R.
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CENTRAL UNIVERSITY OF RAJASTHAN Handling Procedures for Oxidizers

Always handle in a chemical fume hood. Wear safety goggles, a lab coat, and appropriate gloves. Use fire blankets to deprive fires of oxygen Shut sash on fume hood fires Health & Environmental C U RajasthanSafety 32

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Flammables

Flammable Liquid Any liquid that has a flash point below 100oF

Differs from combustible liquid (Any liquid that has a flash point
at or above 100oF)
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Flammables

Flammable Solids Any solid that catches fire through friction or which could be readily ignited.
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While working with flammables

Remove Ignition Sources (heat or spark) Keep away from oxidizers and combustible materials Work in a fume hood or well ventilated area
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Reactive chemicals

Reactive materials can release heat or a toxic or flammable gas upon contact with water. Safe handling of these materials will depend on the specific materials and the conditions in which this material is handled.

Examples (Sodium metal, Lithium aluminum hydride)


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Pyrophoric materials

Pyrophoric materials can react with air and ignite spontaneously at or below 113F. Pyrophoric materials should be handled and stored in inert environments. Examples: (Silane, White or yellow phosphorous, t-butyl lithium)

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Poison/ Toxin

Cause illness or injury by upsetting biological functions or damaging biological structures


Dose-response relationship

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Working with Toxins/ Poison

Read MSDS for exposure limits and proper handling information Use Fume Hood Minimize exposure by using proper protective equipments No food or drinks in lab
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Carcinogenic chemicals

Cause abnormal growth of tissue May have variety of different mechanism Should be handled only after appropriate training Ethidium Bromide, Aromatic Pesticides, etc.
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Radioactive

1. Radioactive atoms with unstable nucleus loses energy by emitting ionizing particles 2. Several applications in laboratory 3. Extremely harmful, know n carcinogen 4. Should be used only after detailed training
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CENTRAL UNIVERSITY OF RAJASTHAN Understand the lab chemical and its potential hazard
Manufacturer Identity Hazards

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National Fire Protection Association (NFPA) Explanation Guide for Chemical hazard

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MSDS Information
Material Safety Data Sheet provides comprehensive information about: Physical Properties Hazards

First Aid measures


Engineering controls PPE

Must have a copy of MSDS for each chemical in the laboratory for easy access in an emergency
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Why MSDS Information important ?


MSDS Contents 1. Identification of substance 2. Composition/Data on components 3. Hazards identification 4. First aid measures 5. Fire fighting measures 6. Spill cleanup measures 7. Handling and storage 8. Exposure controls and personal protection 9. Physical and chemical properties 10. Stability and reactivity 11. Toxicological information 12. Ecological information 13. Disposal considerations 14. Transport information 15. Regulations 16. Other relevant information

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Guidelines for safe storage of laboratory chemical


Must be followed to:
1. Prevent unwanted reactions e.g. incompatible chemicals being mixed
2. Such reactions may generate heat/ fume/ gases that can lead to explosion/ fire and cause damage to personnel and property. 3. Preventing small accident from turning into major disaster

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Chemical Storage Guidelines


The storage area and cabinets should be labeled to identify the hazardous nature of the products stored within

Ensure that all chemicals are stored according to compatibility (e.g., oxidizers away from flammable liquids).
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Flammables storage
1. Flammables include organic liquids, combustibles, and organic acids etc. 2. Flammable materials kept outside a cabinet should be in fire proof safety cans. 3. Flammable liquids (>1L) must be stored inside flammable cabinets. 4. Never store flammable liquids in a standard cabinet. Example: i.e. acetone, hexane, acetic acid, phenol, and chloroform

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1. Solvents should be stored in well organized dedicated storage cabinet

2.

Read MSDS carefully to determine compatibility before storage

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Acids Storage Store by acid class in separate secondary containment Never store acids above eye level

Concentrated per-chloric acid should be stored alone in a cabinet by itself


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Bases Storage Store in secondary containment

Store away from acids and solvents


Never store at or above eye level. Examples: Hydroxides Amines

Ammonia
Bleach
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Dry non-hazardous chemicals can be stored on laboratory shelves.

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Chemicals should never be stored on the floor. If need be they must be stored in approved secondary containers.

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Compressed Gases
Compressed gases may pose a physical and/or health hazard depending on the gas being used. Restraint:

Gas cylinders must be individually secured with suitable regulators


Only cylinders with safety caps can be group chained.

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CENTRAL UNIVERSITY OF RAJASTHAN Storing compressed gases


Flammable gases (i.e. propane, hydrogen) must be stored away from sources of ignition and oxidizing gases (i.e. oxygen).

Poison/toxic gases (i.e. chlorine, carbon monoxide, phosgene or ammonia) can pose serious potential hazards to personnel and therefore special storage and handling measures are required.
Poison gases must be stored as follows:
Small size cylinders (fit inside hood) Use and storage in hood Large size cylinders (regular use) Use and store in ventilated
with an air monitoring and alarm system

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Design and working principles of Fume Hoods

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Using Fume Hoods


Use For All Operations Where Odoriferous, Volatile, Toxic or Harmful Release Possible

When using the fume hood you first make sure that the exhaust blower is operating and air is entering the hood.
Remember, do not put your face inside the hood

Work at least 6 inches into the hood


Minimize storage of chemicals in the hood Clean spills immediately Work with the sash at the proper operating level as guided by the indicator arrows
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Testing Fume Hood Functionality

1. 2.

Functional testing of the Fume Hoods is done with Dry Ice Fume Hoods with unidirectional blow of dry ice away from the sash
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Biological Hazards in a laboratory

Several forms of biological hazards


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CENTRAL UNIVERSITY OF RAJASTHAN Before you use any biological sample in the laboratory
Learn about the potential risk associated with any biological sample before using it in your lab experiment

(Sources for information)

Center for Disease Control (CDC) Food & Drug Administration (FDA) American Type Culture Collection (ATCC) German Culture Collection (DSMZ) Microbial Type Culture Collection (MTCC) C U Rajasthan- Health & Environmental
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Information about the bio-safety level of biological sample

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Aerosols and Droplets


Aerosols and Droplets are the main sources of biohazard contamination

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Reduce Aerosols
Gently expel fluids against the walls of tubes or flasks Perform aerosol forming experiments in Biological Safety Cabinet
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Bio Safety Level I
Hazard group 1- P1 Doors closed during work Normal laboratory procedures

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Containment level 2
Hazard group 1 2 (P1- P2) Use Class II safety cabinet Use autoclave Protect against aerosol generation

Class II

Auto

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Bio Safety Level III
Hazard group 3 P1 -P3 Keep Laboratory and changing room at negative pressure so air flows from outdoors to indoors Use Class II safety cabinet

Entry

Change room ( ve)

ve) Class II Minus pressure

Auto

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Bio Safety Level IV
Hazard group 4, P1 -P4 Keep rooms at negative pressure Use Class III safety cabinet Optional air inlet Install airlock and shower
Restricted entry Change room Minus pressure

-ve
Shower

-ve

-ve

-ve
Class III

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Laminar Air Flow (LAF)
Unidirectional straight uninterrupted airflow Sufficient for working with BSL1 agents Generally used where protection is required for product/ work only

No protection to operator

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Design and working principles of Bio Safety Cabinets
Bio Safety Cabinet Class I
1. Unidirectional air flow from working area towards HEPA filter aided exhaust

2.
3.

Some protection to user & environment


No protection to worked

Bio Safety Cabinet I


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Design and working principles of Bio Safety Cabinets
Bio Safety Cabinet Class II
1. Bi-directional flow of HEPA filter purified air

2.

Good protection to user, the environment and the work

Bio Safety Cabinet II


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Design and working principles of Bio Safety Cabinets
Bio Safety Cabinet Class III
Fan Anti blow-back valve + alarm Exhaust HEPA Pre-filter Inlet HEPA

1. 2. 3. 4.

Generally used for the highest hazard category Fully sealed shell operator protection for

Work is done by using gloved sleeved ports Air is drawn into and out of the cabinet via HEPA filters

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Clean Area

Working Area

Contaminated Area
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Never block the rear vents of the Bio-safety Cabinets

It defeats the basic principle of the BCSs.


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Store required supplies outside of Bio-safety Cabinets

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Never use LAF/ BSCs as a substitute to fume hoods

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Keeping proper information/ record of the BSL Laboratory
BSL-1 Laboratory
Information sheet must contain

Name of infectious agent(s)


Mode of transmission(s)
BIOHAZARD
All Personal Protective Equipment shall be removed prior to leaving this work area. Eating, drinking, smoking applying cosmetics or lip balm and handling contacts lenses area prohibited in this work area. Name of infectious agent(s):
__________________________________________________________________________________ __________________________________________________________________________________

Requirements of entering lab Emergency Contact Person, Phone

Transmission route(s):
__________________________________________________________________________________

Special requirements for entering this area:


__________________________________________________________________________________
__________________________________________________________________________________

Emergency contacts Name: ________________________________ Title: ______________ Phone: _______________


________________________________ ________________________________ _______________ _______________ _______________ _______________

EH&S office

(M-F 8-5) 646-3327 (after hours) 911.

__________________________________________________________________________________ *Biosafety Level 2 is similar to BSL-1 and is suitable for work involving agents of moderate potential hazard to personnel and the environment. It differs from BSL-1 in that 1.Lab personnel have specific training in handling pathogenic agents and directed to competent scientist; 2. Access to the lab is limited when work is being conducted: 3. Extreme precautions are taken with contaminated sharp items and 4. Certain procedures in which infectious aerosols or splashes may be created are conducted in biological safety cabinets or other physical containment equipment
New Mexico State University Environmental Health & Safety Date _______

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Used injection and sharps are the other common source of bio-hazard contamination

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Scissors, scalpels, razor blades, pins, syringe needles and knives are sharp so use care. Always direct sharp edges and points away from yourself and others. If you are cut or bleeding, let your teacher know immediately. Use only as directed.
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Safe handling of needles and sharps

Needles must not be recapped, bent, sheared or removed from a disposable syringe. Used needles should be destroyed. All used sharps must be placed in a rigid, hard-plastic, punctureresistant container for disposal.
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Safe handling of broken glass-ware

Broken glassware within laboratory should not be handled with bare hands

Broken glassware should be disposed properly


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Safety Supplies and Equipments

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Common Laboratory Safety Equipment

Regular Safety checks

Fire safety

Personal protective equipment

Lab shower Eye wash station

Waste disposal
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Personal protective equipment (PPE)


Includes any devices or clothing worn by the worker to protect against the hazards in the environment. Examples Safety glasses

Safety masks
Lab coats & Shoes Protective gloves,

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Protective Laboratory Goggles/ Masks

Put on your safety glasses and mask before entering the lab. You never know what may come flying off of a work bench, a fume hood, or a refrigerator Use of protective masks minimizes the chances for inhalation dependent exposure
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Protective Clothing (Lab Coat)


Must covers arms and legs

Lab coats with closed fasteners


Non-flammable, non-porous aprons when using corrosives Remove before leaving the lab Launder separately

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Lab coat, goggles and glove practice

Inappropriate

Appropriate
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Personal protection before entering the lab


Minimize exposed skin at all times by wearing the appropriate clothing Use appropriate protective equipment at all times in the lab when chemical, radioactive, or bio-hazardous materials are being handled.

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Emergency Laboratory Shower

1.

Each lab should be equipped with an emergency shower at an unobstructed and easily accessible location within the laboratory preferably within 10 seconds Laboratory showers pour water at ~ 20 gals. / min. (compare to 2 gals. / min.) of standard bathroom shower
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2.

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Eye Wash Station

Each lab should be equipped with an eye wash station

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How to use an Eye Wash Station
Shout for help Hold your eyelids open with your fingers as you walk towards eye wash unit Rinse your eyes thoroughly Ask a co-worker to watch clock to make sure that you continued to rinse your eyes for at least 15 minutes Ask a co-worker to collect MSDS and visit a doctor
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CENTRAL UNIVERSITY OF RAJASTHAN Personal protective equipments


Many types of
1. 2. 3. Chemical Fume Hoods Laminar Air Flow Bio-safety Cabinets

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1. Fire extinguishers are important means of laboratory safety.

2. Should be used as per recommendations and only after appropriate training.


3. Never use fire extinguisher for large fires
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CENTRAL UNIVERSITY OF RAJASTHAN Other Safety Equipments

1. First Aid Kit should be accessible to all workers in the lab 2. First Aid Kits should be stocked and maintained regularly 3. Laboratory workers should be trained for use of First Aid Kit
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Safe Laboratory Practices

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5. Follow rules & guidelines


Dos

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Donts

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Floor- plans with fire exit procedure

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Safety from fire

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Fire: The other very common lab hazard Fire Safety


All employees are responsible for safety. Know the location of fire extinguishers .

Learn how to use them correctly.


Know where the fire blanket is. Know the procedure for reporting a fire.

Institution/ University should organize fire safety training/ drills.


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Fire Safety
Components of a fire
Fuel Oxygen

Heat
Necessary chain reaction

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Fire Safety
Classification of fires
1. Class A fires

Occur with ordinary combustible material, such as wood, rubber, paper, cloth, and many forms of plastics.
2. Class B fires

Occur in a vaporair mixture over flammable solvents, such as gasoline, oil, paint, lacquers, grease, and flammable gases.
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Fire Safety
Classification of fires
3. Class C fires

Occur in or near electrical equipment.

4.

Class D fires Occur with combustible metals, such as magnesium, sodium, and lithium.

(Class D fires should be fought by firefighters only)

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Courtesy of Health and Environmental Safety, The University of Texas Health Sciences Center at Houston

Fire Safety

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Fire Safety
Fire Extinguishers
1. Type A extinguishers
Contain soda and acid or water and are used to cool the ordinary fire such as wood, cloth or paper.

2.

Type BC extinguishers
Contain foam, dry chemicals, or carbon dioxide (CO2). Are used to combat fires occurring in vaporair mixtures over solvents such as grease, gasoline or oil fires.

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3. Type ABC extinguishers
Contain a dry chemical and are used on fires of wood, cloth, paper, oil, grease, and gasoline. Multipurpose in combating fires and thus, these are best suited for use in research laboratories.

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Fire fighting with Fire extinguishers

P. A. S. S

P = pull pin A = aim at fire S = squeeze lever S = swipe side to side

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Heat Hazard in the laboratory


Use an oven mitt or gloves or clamp when handling hot materials. Always heat objects in a test tube holding away from you and others. Hot plates, hot water and hot glassware can cause burns. Glassware looks the same when hot or cold, so use care when heating any type of glass. Rapid changes in glass temperature can cause it to expand, crack and/or explode! Never touch a hot plate, oven trays etc. with bare hands.

CENTRAL UNIVERSITY OF RAJASTHAN Emergency Response to Possible Fire


Things to do in a fire response 1. Pull the nearest fire alarm 2. Close windows and doors (minimize O2 influx) 3. Use an ABC type extinguisher for small fire 4. If fire is too big, leave the area immediately by stairs 5. Drop to ground and roll or crawl to exit

C U Rajasthan- Health & Environmental Safety

CENTRAL UNIVERSITY OF RAJASTHAN

Your contribution towards making labs safe

Phlebotomy Handbook: Blood Collection Essentials, Seventh Edition Diana Garza Kathleen Becan-McBride

CENTRAL UNIVERSITY OF RAJASTHAN

Students and researchers:


Complete Laboratory Safety Training Ensure complete compliance with the laboratory safety training Ensure own safety and of others

C U Rajasthan- Health & Environmental Safety

110

CENTRAL UNIVERSITY OF RAJASTHAN


How do we ensure safety for all C U Raj. family members ?

University Safety Training Programme


Teach What! (PPT modules and Reading Materials) Train- How! (Hands-on Training & Demos)

Test (Online safety Quiz)

CENTRAL UNIVERSITY OF RAJASTHAN