LABORATORY SAFETY

What does laboratory safety

cover?

Types of hazards, and precautions taken to

protect persons from laboratory hazards

through control of the working environment,

working methods & equipment

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 What are laboratory hazards?

• Chemical hazards
• Electrical hazards
• Physical/mechanical hazards
• Radiation hazards
• Fire hazards
• Biological (infection) hazards

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 Who are at risk?

• Laboratory staff
• Receptionists
• Laboratory cleaners
• Patients
• Visitors

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 Sources of chemical hazards

• Chemical disinfectants
• Volatile chemicals
• Substrate tablets for ELISA
• Other laboratory reagents, e.g. acids,
strong alkalis, poisons

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 Precautions against chemical
hazards
• Don'ts
- No mouth pipetting
- No tasting of chemicals
- Never add water to acid
- Never store corrosive
chemicals in plastic/rubber WATER
containers

ACID

NEVER ADD WATER

TO ACID 6
 Precautions against chemical
hazards
• Do’s
- Understand the standard
commercial warning signs
- Each bottle or container must be ACID
labelled with at least:
- name of reagent
- date of preparation
- Always add acid to water –
never water to acid WATER

- Poisons: must be kept in

lockable cupboards at all times ALWAYS ADD ACID TO WATER

- Open windows to improve

ventilation when handling volatile
chemicals 7
 Precautions against electrical
hazards
• Keep electrical equipment clean and dry, and
away from all water sources
• Connect electrical equipment to fused plugs
• Do not overload electrical outlets
• Do not use faulty equipment.
– Label faulty equipment clearly until repairs are done
• Replace worn out wires promptly
• Major repairs must be handled by authorised
personnel (qualified electricians)

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 Sources of physical/mechanical
hazards
• The following are not recommended for a
laboratory:

– Over crowding
– Polished floor
– Improper furniture
– Improper use of equipment

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 Equipment that may cause
hazards
• Cold storage tanks
• Autoclaves
• Centrifuges
• Mechanical shakers
• Vortex mixers

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Equipment/devices that help to
prevent hazards
• Bio-safety cabinets
• Pipetting aids
• Centrifuge caps
• Gloves
• Goggles
• Masks

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 Biological safety cabinets

• Protection is provided by:

Directional airflow
Hepa filters: High Efficiency Particulate Air
filters
• HEPA filters trap particles of ≥ 0.3µ
• Chemicals, fumes, vapours may
damage HEPA filters
• Expensive and not suitable for small
laboratories
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 Bio-safety Cabinets:
Classification
Three general classes available
• Class 1
• Class II
• Class III

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 Class I BSC
- Protection of worker: inward
airflow
- Protection of environment:
exhaust to outside through
HEPA filter
-No protection of research
materials: air in cabinet not
sterile
-Designed for low and
moderate risk microbiological
agents

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 Class II BSC
-Protection of worker:
inward airflow

-Protection of environment:
exhaust to outside through
HEPA filter

- Protection of research
materials: downward
laminar flow of HEPA-
filtered air provides product
protection

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Biological
safety
cabinet
Type II

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 Class III BSC
-Provides the highest level of
product, environmental, and
personnel protection

-Cabinet is gas-tight with non-

opening view window

-Rubber gloves are attached to

ports for manipulation of
materials in the cabinet

-Air is filtered through one

HEPA filter before entering the
cabinet, and through 2 HEPA
filters before it is exhausted to
the outdoors
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 Protective wear
• Hand gloves
• Eye shield

• Respiratory mask

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 Gloves
•Always wash hands before
wearing gloves
•Always wear the right size of
gloves
•Always wear gloves when handling
body fluids
•Always change gloves between
clients
•Always wear and remove gloves
properly

*** Remember gloves do not protect

from accidental pricks
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 Good Laboratory Practice (GLP)
Safe personal behaviour…

• Put on protective clothing

• Cover wounds or broken skin with water proof
elastoplast
• Do not store food or drinks in the laboratory refrigerator
• Do not smoke, drink or eat in the laboratory
• Do not bite nails, chew pencils or lick gummed labels
• Wash hands with soap and water before leaving the
laboratory or before attending a new patient

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 CONTINUATION (GLP)

• Do not sit on work benches

• Work tidily and avoid cluttering benches
• Do not allow unauthorised persons in the laboratory
• Minimise the use of common hand towels
• Use gloves properly, otherwise they may contribute to
the spread of infection
• Wear laboratory gown/coats only within the laboratory

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 Sources of radiation hazards

• Use of radioisotopes
• Improper disposal
• Improper storage

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 Precautions against fire hazards
• No smoking in the laboratory
• Stopper all flammable chemicals tightly when not in use
• Keep flammable chemicals away from naked flames
• Drain flammable liquids down the sink with plenty of water
• Do not use an open flame against a background of direct sunlight
• Extinguish open flames when not in use
• Handle hot objects with tongs or forceps
• Set up emergency fire-fighting measures at the exit door of the
laboratory:
- Sand bucket
- Fire extinguisher (carbon dioxide type)

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Sources of biological hazards

• Inhalation
• Ingestion
• Contamination of open cuts
• Accidental pricks

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 Precautions against biological
hazards (infection)
• Regard every specimen as potentially infectious
• Flame metal instruments, e.g. wire loops, forceps, at
arm’s length
• Do not recap needles to avoid accidental pricks
• Cap infective fluids securely before centrifuging
• Empty broken glass from centrifuge buckets into a
container of disinfectant and not into the sink

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Never place needles or sharps in
office waste containers!!

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Do’s and Don’ts: Sharps and
Waste Containers

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 Post-HIV Exposure Prophylaxis
(PEP)…
In case of needle PRICK:

1. Wash site with soap and water

Do not squeeze
Do not scrub

2. Record in incident/accident log book

3. Counsel the recipient immediately & perform a rapid HIV test. The
recipient must be HIV negative to benefit from PEP

4. Initiate ARV prophylaxis immediately (1 – 2 hrs) & take for 4 weeks

(can reduce HIV risk by 80%)

Note: The risk of HIV transmission from a single needle stick is

only 0.3%
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 …PEP
Assess risk exposure:

Low risk - Small volume of blood

exposure - Injury with a solid needle
- Superficial injury
High risk - Large volume of blood or infectious fluid
exposure - Contamination from HIV +ve patient with high viral titre
- Injury with hollow needle
- Deep or extensive injury

Treatment of risk category:

Risk category ARV prophylaxis Duration

Low risk Retrovir 200 mg tds (300 mg bd) 28 days

High risk Retrovir 200 mg tds (300 mg bd) 28 days
Epivir 150 mg bd +
Indinavir 800 mg tds
or
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Combivir 1 bd + Indinavir 800 mg tds
 …PEP
5. If HIV status of source person is negative:
• Consider window period
• Repeat HIV test after 2 weeks
• If HIV –ve , reassure, PEP can be discontinued

6. If HIV status of source person is known HIV +ve, proceed with

full course PEP according to risk category

7. Recommended laboratory tests after exposure:

Baseline: full blood count, LFTs, RFTs, HIV serology
Two weeks: full blood count, LFTs & RFTs
Six weeks: HIV serology

8. If HIV +ve, at 6 weeks, discuss with Senior Clinician

NOTE: Non-protective casual sex is considered to be HIGH
RISK EXPOSURE 30
 Cleaning and Washing:
purpose
1. Reduce the number of bacteria contaminating an article.
The article must be dried after cleaning or washing to
avoid multiplication of surviving bacteria

2. Remove dirt, grease or organic matter, e.g. blood, faeces

from an article. The presence of organic matter interferes
with disinfection and sterilisation procedures

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 Disinfection

• Disinfection is the removal of some or all

pathogenic organisms from an article or
surface

• Disinfection does not always destroy

spores, e.g. tetanus spores

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 Disinfection methods

1. Boiling in water

• Water boils at 100°C at sea level. At

higher altitudes, water boils at less than
100°C
• The temperature of boiling water is
reduced by approximately 1°C for every
500 metres above sea level
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 Disinfection

• A disinfectant is a chemical capable of killing

micro-organisms

• An antiseptic is a weak disinfectant. Antiseptics

may inhibit the growth of organisms without
actually destroying them

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 Chemical disinfectants
• Alcohols, e.g. methyl alcohol, ethyl alcohol,
isopropyl alcohol

• Phenols, e.g. Lysol

• Hypochlorite, e.g. household bleach, JIK

• Aldehydes, e.g. formalin, glutaraldehyde

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 How do chemical disinfectants
work?
• Protein denaturisation and precipitation, e.g. alcohols,
phenols, aldehydes
• Disruption of cell membranes, e.g. bleach
• Factors determining the effectiveness of chemical
disinfectants:
- concentration of chemical
- time of application
- frequency of change
- presence of inactivating materials, e.g. organic materials
(blood, stool)
- pH
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 Chemical disinfection:
criteria for choosing disinfectants
• Mode of action
- Cidal
- Static

• Rate of action
- Concentration

• Side effects
- Corrosiveness
- Irritant vapours
- Staining properties

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 Common disinfectants: dilutions, activity, and
potential applications
Disinfecta Working solution Activity Routine use
nt
Alcohols 70% Kills a wide range of Skin disinfectant
vegetative bacteria
and HIV. Does not kill
bacterial and fungal
spores
Phenols 5% Lysol® Kills a wide range of Re-usable glassware e.g. slides, tubes
vegetative bacteria etc. 10% Lysol® for body fluids
2% Lysol®, and some viruses Washing hands
Sudol®, Hycolin® after 30 minutes.
Does not kill bacterial
spores
Halogens 0.5% hypochlorite Kills viruses, General hospital disinfectant of choice for
solution vegetative bacteria, gloves, spillage, broken glass
spores and fungi Do not use on metal instruments
Note: Prepare freshly each day.

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0.1% hypochlorite Washing hands
0.25% iodine Skin disinfectant
 Sterilisation

• Process of rendering an item free from all

living micro-organisms

• All bacteria, fungi, spores and viruses are

killed during sterilisation

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 Sterilisation methods

• Flaming
• Filtration
• Hot air oven (dry heat)
• Autoclaving (steam under pressure)
• Incineration

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 What sterilisation methods are
used at Health Centre and District
Hospital levels?
What sterilisation methods are used
at Health Centre and District
Hospital levels?

• Flaming
• Autoclaving (steam under pressure)
• Incineration

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 Steam under pressure
The autoclaving cycle

121oC
15 min
100oC A = boiling
B = air expulsion
C = pressure building
D = holding time
A B C D E
E = cooling

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 Indicators of sterilisation

• Autoclave tape

• Browne’s indicator tube

• Time Steam and Temperature Tube

• Biological testing, i.e. use of

Bacillus stearothermophilus
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 Guidelines for waste disposal
Develop a policy for disposal of medical waste

• Divide used materials into two types:

- Re-usable
- Disposable = WASTE
• Segregate infectious waste from other waste, e.g. papers,
liquid waste, sharps, etc
• Sterilise or disinfect contaminated/infectious waste before
it leaves the laboratory
• Seal infectious waste to prevent leaks and spills
• Appoint someone to oversee disposal procedures
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 Waste disposal
Does your waste
disposal system
look like this?

Infectious laboratory waste is a risk to

health care workers and the community
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 Incineration

• Incineration is the complete burning of material to

ashes. This is the most effective method of
destroying infected disposable material

• What materials should be incinerated?

- All burnable materials including cotton gauze,
cotton wool, syringes, plastic containers, etc

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 How to use a simple incinerator
1. Place the material in the incinerator

2. Sprinkle fuel on the material, e.g. diesel, kerosene or

spirit, and light. Close the incinerator door so the
material burns at low oxygen levels

3. Once a week remove the ashes and other debris from

the incinerator and dispose of into a deep pit. The pit
should be placed near the incinerator

The incinerator and pit must be fenced to prevent

access by unauthorised persons
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 Key Messages
• Safety is primarily a personal responsibility, but
training and orientation are paramount

• It is important to understand the sources of

laboratory hazards

• Waste must be categorised and appropriate

protocols for disposal employed

• Accidents/incidents need to be documented and

appropriate action taken
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Any questions?
Safety references

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Thank you!