General Lab Safety

CENTRAL UNIVERSITY OF RAJASTHAN

Laboratory Safety Training

Dr. Janmejay Pandey

Central University of Rajasthan


Outline of the presentation:

1. Why is it important

2. Some fundamental Concepts

3. Safety in Chemical & Biological Labs

4. Safety Supplies and Equipments

5. Safe Laboratory Practices

6. Safety from fire

7. Your contribution towards making labs safe

Central University of Rajasthan


C U Rajasthan- Health & Environmental

Safety 3

Why is it important ?


C. U. Rajasthan- Motto

Education for sustainable Development

Laboratory Safety !

A critical component of sustainable development


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.

5 C U Rajasthan- Health & Environmental

Safety



Safety 6

Negligence of lab safety : A horror story

1. UCLA (Dec. 18, 2008). A chemistry researcher working with a pyrophoric chemical got caught in a Laboratory fire.

2. She suffered deep 3rd degree burns over 40 % of her body & 18 days later she died from this accident.

Sheri Sangji



Safety 7

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.

2. The bottle slipped out of her hand, content of the bottle spilled over her. It turned out to be concentrated acid.



Safety 8


Do we need to be in accidents to learn lessons !



Safety 10

Most frequently reported infections in Laboratory workers during last 10 years

Disease or Agent No. of Cases

Mycobacterium tuberculosis 223

Q fever 176

Hantavirus 169

Hepatitis B virus 84

Brucella sp. 81

Salmonella sp. 66

Shigella sp. 56

Hepatitis non-A, non-B 28

Cryptosporidium sp. 27

Total 1074



Safety 11

Flying laboratory instruments/ objects



Safety 12

Injuries to Research Students/ Employee

Source: Howard Hughes Medical Institute, Office of Laboratory Safety



Safety 13

Source: Howard Hughes Medical Institute, Office of Laboratory Safety

Injuries to Laboratory Support Staff/ Random Visitor


1. Regulatory requirement in several developed/ developing countries and organizations e.g. (NIH, CDC, OSHA, EPA- USA), (CSIRO- Aus), (EMBL- EU).

2. Institutional and Laboratory quality assessment & audit.

Training is an important government regulation in western world

Government Regulations for Lab Safety


Safety



Safety 15

Indian Status


• 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 !


Safety

Lab Safety Measures



Safety 17

Key Concepts of Laboratory Safety

Laboratory safety revolves around

understanding, safe storage, usage & disposal of

hazardous components of lab work



Safety 18

Key Concepts of Laboratory Safety

1. Understand

2. Organize

3. Ensure containment

4. Protect

5. Be prepared



Safety 19

1. Understand

Prior preparation is extremely useful in ensuring lab safety.

Physical: Get training for safe use of instrument.

Chemical: Use MSDS and other source about chemicals.

Biological: Read about hazardous effects of biological material.

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



Safety 20

• Organize your work and work place before you start.

• Work diligently.

• Be alert and attentive.

• Don’t 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

2. Organize



Safety 21

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.


4. Protect (yourself and co-workers)

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

Boots etc.).

2. Chemical Fume Hood.

3. Laminar Air Flow and Bio-Safety Cabinets.

4. Protective Working Shield.



Safety 23

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.



Safety 24

Safety in Chemical & Biological Labs



Safety 25

Chemical hazards in a laboratory

Several forms of chemical hazards



Safety 26

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

Explosive



Safety 27

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



Safety 28

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



Safety 29

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



Safety 30

(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.



Safety 31

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.



Safety 32

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



Safety 33

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)



Safety 34

Flammables

Flammable Solids

• Any solid that catches fire through friction or which could be readily ignited.



Safety 35

• Remove Ignition Sources (heat or spark)

• Keep away from oxidizers and combustible materials

• Work in a fume hood or well ventilated area

While working with flammables



Safety 36

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)



Safety 37

• Pyrophoric materials can react with air and ignite spontaneously at or below 113°F.

• Pyrophoric materials should be handled and stored in inert environments.

Examples: (Silane, White or yellow phosphorous, t-butyl lithium)

Pyrophoric materials



Safety 38

Poison/ Toxin

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

• Dose-response relationship



Safety 39

• 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

Working with Toxins/ Poison



Safety 40

• 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.



Safety 41

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



Safety 42

Manufacturer

Identity

Hazards

Understand the lab chemical and its potential hazard

?



Safety 43

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



Safety 44

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



Safety 45

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

MSDS Contents

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

Why MSDS Information important ?



Safety 46

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

Guidelines for safe storage of laboratory chemical



Safety 47

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).



Safety 48

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



Safety 49

Solvent storage

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

2. Read MSDS carefully to determine compatibility before storage



Safety 50

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



Safety 51

Bases Storage

• Store in secondary containment

• Store away from acids and solvents

• Never store at or above eye level.

• Examples:

• Hydroxides

• Amines

• Ammonia

• Bleach



Safety 52

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



Safety 53

• Chemicals should never be stored on the floor.

• If need be they must be stored in approved secondary containers.



Safety 54

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.



Safety 55

• 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

Storing compressed gases



Safety 56

Design and working principles of Fume Hoods



Safety 57

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



Safety 58

Testing Fume Hood Functionality

1. Functional testing of the Fume Hoods is done with Dry Ice 2. Fume Hoods with unidirectional blow of dry ice away from

the sash



Safety 59

Biological Hazards in a laboratory

Several forms of biological hazards



Safety 60

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)

Before you use any biological sample in the laboratory



Safety 61

Information about the bio-safety level of biological sample



Safety 62

Aerosols and Droplets are the main sources of biohazard contamination

Aerosols and Droplets



Safety 63

Reduce Aerosols

• Gently expel fluids against the walls of tubes or flasks

• Perform aerosol forming experiments in Biological Safety Cabinet



Safety 64

Bio Safety Level I • Hazard group 1- P1

• Doors closed during work

• Normal laboratory procedures



Safety 65

Containment level 2

• Hazard group 1 – 2 (P1- P2)

• Use Class II safety cabinet

• Use autoclave

• Protect against aerosol generation

Auto

Class II


Auto

Class IIMinus

pressure

Change room

( ve) ( ve)

Entry


Safety 66

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



Safety 67

Bio Safety Level IV • Hazard group 4, P1 -P4

• Keep rooms at negative pressure

• Use Class III safety cabinet

• Install airlock and shower

Class III

Minus

pressure

Change roomRestricted

entry

Shower

Optional

air inlet

-ve

-ve -ve

-ve



Safety 68

• Unidirectional straight uninterrupted airflow

• Sufficient for working with BSL1 agents

• Generally used where protection is required for product/ work only

• No protection to operator

Laminar Air Flow (LAF)



Safety 69

Bio Safety Cabinet I

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. Some protection to user & environment

3. No protection to worked



Safety 70

Bio Safety Cabinet II


Bio Safety Cabinet Class II

1. Bi-directional flow of HEPA

filter purified air

2. Good protection to user, the environment and the work



Safety 71

Inlet

HEPA

Fan

Anti blow-back

valve + alarm

Exhaust

HEPA

Pre-filter


Bio Safety Cabinet Class III 1. Generally used for the

highest hazard category

2. Fully sealed shell for operator protection

3. Work is done by using gloved sleeved ports

4. Air is drawn into and out of the cabinet via ‘HEPA’ filters



Safety 72

Biological Safety Cabinet Operation Procedure

Clean Area Working Area Contaminated Area



Safety 73

Never block the rear vents of the Bio-safety Cabinets

It defeats the basic principle of the BCSs.



Safety 74

Store required supplies outside of Bio-safety Cabinets



Safety 75

Never use LAF/ BSCs as a substitute to fume hoods



Safety 76

BSL-1 Laboratory

BIOHAZARDAll 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):__________________________________________________________________________________

__________________________________________________________________________________

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 Date _______

Environmental Health & Safety

Keeping proper information/ record of the BSL Laboratory

Information sheet must contain • Name of infectious agent(s)

• Mode of transmission(s)

• Requirements of entering lab

• Emergency Contact Person, Phone



Safety 77

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



Safety 78

• 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.



Safety 79

• 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, puncture-resistant container for disposal.

Safe handling of needles and sharps



Safety 80

Safe handling of broken glass-ware

Broken glassware within laboratory should not be handled with bare hands

Broken glassware should be disposed properly



Safety 81

Safety Supplies and Equipments



Safety 82

Common Laboratory Safety Equipment

Personal protective equipment

Waste disposal

Fire safety

Lab shower

Eye wash station

Regular Safety checks



Safety 83

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,



Safety 84

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



Safety 85

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



Safety 86

Lab coat, goggles and glove practice

Inappropriate Appropriate



Safety 87

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.



Safety 88

1. Each lab should be equipped with an emergency shower at an un-obstructed and easily accessible location within the laboratory preferably within 10 seconds

Emergency Laboratory Shower

2. Laboratory showers pour water at ~ 20 gals. / min. (compare to 2 gals. / min.) of standard bathroom shower

http://www.gesafety.com/downloads/G1643.pdf

http://www.gesafety.com/products/rec_lab/GBF2120.shtml



Safety 89

Each lab should be equipped with an eye wash station

Eye Wash Station



Safety 90

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

Regular testing and maintenance of Emergency Shower, Eye Wash Station & document test reports



Safety 91

Personal protective equipments

Many types of

1. Chemical Fume Hoods

2. Laminar Air Flow

3. Bio-safety Cabinets



Safety 92

Other Safety Equipments

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



Safety 93

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



Safety 94

Safe Laboratory Practices



Safety 95

Dos

5. Follow rules & guidelines



Safety 96

Don’ts



Safety 97

Floor- plans with fire exit procedure



Safety 98

Safety from fire


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.

Fire: The other very common lab hazard


Safety


Fire Safety

Components of a fire

• Fuel

• Oxygen

• Heat

• Necessary chain reaction


Safety


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 vapor–air mixture over flammable solvents, such as gasoline, oil, paint, lacquers, grease, and flammable gases.


Safety


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)


Fire Safety

Courte

sy o

f Health

and E

nviro

nm

enta

l Safe

ty, T

he U

niv

ers

ity o

f Te

xas H

ealth

Scie

nces C

ente

r at H

ousto

n


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 vapor–air mixtures over solvents such as grease, gasoline or oil fires.


Fire Extinguishers

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.


Safety


Fire fighting with Fire extinguishers

P. A. S. S

P = pull pin A = aim at fire

S = squeeze lever S = swipe side to side


• 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.

Heat Hazard in the laboratory


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


Safety


Phlebotomy Handbook: Blood Collection Essentials, Seventh

Edition

Diana Garza • Kathleen Becan-McBride

Your contribution towards making labs safe



Safety 110

Students and researchers:

• Complete Laboratory Safety Training

• Ensure complete compliance with the laboratory safety training

• Ensure own safety and of others


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)


General Lab Safety

Documents

safety supplies

rajasthan motto education

negligence of lab safety

lab safety measures

lack of lab safety training

laboratory workers

safe laboratory practices

hepatitis non