Biosafety level and biosafety cabinets

Techef In
Biosafety LeVeLs &Biosafety LeVeLs &
Biosafety CaBinetsBiosafety CaBinets
 Bio safety Levels
 Air Filtration in Biosafety cabinets
 Standards
 Hood , Laminar flow & Bio safety Cabinets
 Classification –Bio safety Cabinets
TopicsTopics
Biosafety LevelsBiosafety Levels
 Bio Safety Level (BSL)-I:
Well-characterized agents not known to consistently cause disease in healthy adult
humans, and of minimal potential hazard to laboratory personnel and the
Environment.
Standard practices required:
 frequent hand washing
 door that can be kept closed when working
 limits on access to the lab space when working
 no eating, drinking, storage of food in laboratory
 care to minimize splashes and actions that may create aerosols (tiny droplets)
 decontamination of work surfaces after every use
Primary Safety Barriers- Not Required
Biosafety LevelsBiosafety Levels
 Bio Safety Level (BSL)-II:
• Suitable for work involving agents of moderate potential hazard to personnel
and the environment.
• Generally associated with human disease
Safety Equipment (Primary barriers):
• Class I or II Bio safety Cabinets or other physical containment devices (eg….
Aerosol tight lid in Centrifuges) are used
• Personnel Protection Equipment: Laboratory coats, gloves, face protection as
needed
Facilities (Secondary barriers):
• Provide lockable doors for facilities that house restricted agents
• Locating new laboratories away from public areas.
• Furniture used in laboratory work should be covered with a non-fabric material that
can be easily be decontaminated.
• An eye wash station is readily available.
Biosafety LevelsBiosafety Levels
 Bio Safety Level (BSL)-III:
Applicable where work is done with indigenous or exotic agents which may cause
serious or potentially lethal disease as a result of exposure by the inhalation
route.
Safety Equipment (Primary barriers):
• Class I or II Bio safety Cabinets
• Personnel Protection Equipment: Laboratory coats; gloves; face protection as
needed
• Facilities (Secondary barriers):
• The laboratory has special engineering and design feature . The exhaust air from the
laboratory room is discharged to the outdoors
• The ventilation to the laboratory is balanced to provide directional airflow into the
room
Biosafety LevelsBiosafety Levels
 Bio Safety Level (BSL)-IV:
Dangerous/exotic agents which pose high risk of life-threatening disease, aerosol-
transmitted lab infections or related agents with unknown risk of transmission.
Safety Equipment (Primary barriers):
All procedures conducted in Class III Bio safety Cabinets
or Class I or II Bio Safety Cabinets in combination with full-body, air-supplied, positive
pressure personnel suit.
Facilities (Secondary barriers):
BSL-3 plus:
• Separate building or isolated zone
• Dedicated supply and exhaust and decontamination systems
Biosafety LevelsBiosafety Levels
Air Filtration in Biosafety cabinetsAir Filtration in Biosafety cabinets
•High efficiency particulate air (HEPA) filters
•Ultra low particulate air (ULPA) filters
Air Filtration in Biosafety cabinetsAir Filtration in Biosafety cabinets
Variations in HEPA
filter efficiency from
99.95 % to 99.99 % are
usually due to
manufacturing
techniques.
Depending upon its quality, a HEPA
filter is able to trap from 9,995 to 9,999
of every 10,000 particles
An ULPA filter is able to
trap from 99,995 to 99,999 of
every 100,000 particles
ULPA - this is at least 10 times
better than HEPA filtration !!!
International Standards on Biosafety CabinetInternational Standards on Biosafety Cabinet
Why a standard ?Why a standard ?
International Standards on Biosafety CabinetInternational Standards on Biosafety Cabinet
International Standards are mainly issued by two Organisations:
International Standard Organisation (ISO), based in Geneva
(Switzerland),
Comitee Europeen de Normalisation (CEN) based in Brussels
(Belgium).
These standards replace all other local standards on the same
subject.
The aim of the standards is to construct a mechanism to provide
manufacturer and user of a common site to specify technical
specifications and features.
Both construction and testing of the cabinet are covered
International Standards on Biosafety CabinetInternational Standards on Biosafety Cabinet
Standards:
BS-5726 (U.K.)
NF X44-201 (France)
DIN 12950 (Germany)
Now Replaced by EN 12469
EN 12469
European Standards on Design, Construction & Testing of Bio safety Cabinets
In addition to comply DIN 12980 for safety handling of cytotoxic drugs
NSF-49 (National Sanitation Foundation)
In the USA the NSF-49, was one of the first standards on this subject and was the
reference point for a lot of manufacturers. NSF is an independent organisation that
works as neutral agency to help on the problems that could affect to public health
and environment protection.
Was published in 1976.
Difference between Hood and Safety CabinetDifference between Hood and Safety Cabinet
Exhaust of
vapors and
aerosols from
Laboratory to
outside
No validatable
product or
personnel
protection
Validatable
personnel, product
& environmental
protection
Filters and
blower(s)
incorporated
HoodHood Safety CabinetSafety Cabinet
A fume hood is a ventilation device that is designed to
limit exposure to hazardous fumes vapors or dust. A fume
hood is typically a large piece of equipment enclosing
five sides of a work area, the bottom of which is most
commonly located at a standing work height.
Two main types exist, ducted and recalculating. The
principle is the same for both types: air is drawn in from
the front (open) side of the cabinet, and either expelled
outside the building or made safe through filtration and
fed back into the room.
Fume HoodFume Hood
Fume Hood
Only for Product Protection
No Personnel or Environment Protection
Laminar FlowLaminar Flow
A. front opening
B. sash
C. supply HEPA filter
D. blower
Vertical Laminar Flow
Laminar FlowLaminar Flow
A. front opening
B. supply grille
C. supply HEPA filter
D. supply plenum
E. blower
F. Grille
Horizontal Laminar Flow
Classification of Bio Safety CabinetClassification of Bio Safety Cabinet
Biological Safety Cabinet
Class I Class II
Class
III
Type A Type B
Type A
I
Type A
II
Type B
I
Type B
II
Classification of Biosafety CabinetClassification of Biosafety Cabinet
Class I:
Personnel and Environment protection.
Product is not protected.
Class II:
Personnel, Product and Environment are protected.
Class III:
Maximum protection to Personnel, Product and Environment.
Classification of Biosafety CabinetClassification of Biosafety Cabinet
A. Front opening
B. Sash
C. Exhaust HEPA
Filter
D. Exhaust plenum
Class I Biosafety Cabinets
 This is a negative-pressure,
ventilated cabinet usually
operated with an open front.
 All of the air from the cabinet is
exhausted through a HEPA filter
either into the laboratory or to
the outside.
 The Class I BSC is designed for
general microbiological research
with low- and moderate-risk
agents.
Classification of Biosafety CabinetClassification of Biosafety Cabinet
A. front opening
B. sash
C. exhaust HEPA filter
D. rear plenum
E. supply HEPA filter
F. blower
Class II Type A-I Biosafety Cabinets
 It is designed with inward air
flow to protect personnel,
Product & Environment
 HEPA-filtered downward
vertical laminar airflow for
product protection, and HEPA-
filtered exhaust air for
environmental protection
Classification of Biosafety CabinetClassification of Biosafety Cabinet
 Face Velocity 75 fpm
 70% re circulated through HEPA Filter
 30% Exhausted Through HEPA Filter
 Cannot be used for Radio Nucleotides and Toxic Chemicals
 Biosafety Levels 2 & 3
 User, Product, & Environment Protection
Class II Type A-I Biosafety Cabinets
Classification of Biosafety CabinetClassification of Biosafety Cabinet
Class II Type A-II Biosafety Cabinets
A. front opening
B. sash
C. exhaust HEPA filter
D. supply HEPA filter
E. positive pressure plenum
F. negative pressure plenum
Note: The cabinet exhaust
needs to be connected to the
building exhaust system.
Classification of Biosafety CabinetClassification of Biosafety Cabinet
Class II Type A-II Biosafety Cabinets
 Downward Velocity 100 fpm
 70% re circulated through HEPA
 Filter30% Exhausted Through HEPA Filter
 Can be used for Radio Nucleotides and Toxic Chemicals
 Biosafety Levels 2 & 3User, Product, & Environment Protection
 Under Negative Pressure to room
 Exhaust Air is Hard Ducted
Classification of Biosafety CabinetClassification of Biosafety Cabinet
Class II Type B-I Biosafety Cabinets
A. front opening
B. Sash
C. exhaust HEPA filter
D. supply plenum
E. supply HEPA filter
F. Blower
G. negative pressure exhaust plenum
Note: The cabinet exhaust needs to be
connected to the building exhaust
system.
Classification of Biosafety CabinetClassification of Biosafety Cabinet
Class II Type B-I Biosafety Cabinets
 Downward Velocity 100 fpm
 30% re circulated through HEPA Filter
 70% Exhausted Through HEPA Filter
 Can be used for Radio Nucleotides and Toxic Chemicals
(Low Levels / Volatility)
 Biosafety Levels 2 & 3User, Product, & Environment
Protection
Classification of Biosafety CabinetClassification of Biosafety Cabinet
Class II Type B-II Biosafety Cabinets
A. front opening
B. Sash
C. exhaust HEPA filter
D. supply HEPA filter
E. negative pressure exhaust plenum
F. supply blower
G. filter screen
Note: The carbon filter in the building
exhaust is not shown. The cabinet
exhaust needs to be connected to
the building exhaust system
Classification of Biosafety CabinetClassification of Biosafety Cabinet
Class II Type B-II Biosafety Cabinets
 Downward Velocity 100 fpm
 No Re-circulation100% Exhausted Through HEPA Filter
 Can be used for Radio Nucleotides and Toxic Chemicals
 Biosafety Levels 2 & 3User, Product, & Environment
Protection
 Exhaust HEPA Filtered
 Exhaust Hard Ducted out.
Classification of Biosafety CabinetClassification of Biosafety Cabinet
Class III Biosafety Cabinets
A. glove ports with O-ring for attaching
arm-length gloves to cabinet
B. Sash
C. exhaust HEPA filter
D. supply HEPA filter
E. double-ended autoclave or pass-
through box
Note: A chemical dunk tank may be
installed which would be located
beneath the work surface of the BSC
with access from above. The cabinet
exhaust needs to be connected to the
building exhaust system.
Classification of Biosafety CabinetClassification of Biosafety Cabinet
Class III Biosafety Cabinets
 This is a totally enclosed, ventilated cabinet
 Gas-tight construction
 Offers the highest degree of personnel and environmental protection
from infectious aerosols as well as protection of research materials
from microbiological contaminants.
Cytotoxic Drug Safety CabinetsCytotoxic Drug Safety Cabinets
Hepa filter exhaust #2
Hepa filter (pre-filter) #1
Laminar flow filter
Carbon filter exhaust #3 (option)
Dedicated exhaust flow
blower
All contaminated areas under
NEGATIVE pressure
Dedicated Laminar flow
blower
70% re circulated HEPA
filtered laminar flow
Provides sample
protection
30% inflow provides operator
Safety barrier
Can you see the problems ?Can you see the problems ?
Window
Air vent
Ceiling clearance
or wall clearance
Front
Venting
Exhaust
BEWAR
E
Location In LabLocation In Lab
Location A, E and F
are well selected.
As well influencing
air flows through
the Laboratory
personnel traffic is
avoided.
Location B is often
influenced from air
flows in the Room.
Location C is
influenced from
personnel traffic
and Room air
flows.
Location D is also very often influenced
from personnel traffic and air flows.
Safety Cabinets should only be
placed in quiet areas!
Only validated tests can show placement in the
laboratory is correct.
In this instance the tests are invaluable!
THANK YOUTHANK YOU
1 sur 34

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Biosafety level and biosafety cabinets

  • 1. Biosafety LeVeLs &Biosafety LeVeLs & Biosafety CaBinetsBiosafety CaBinets
  • 2.  Bio safety Levels  Air Filtration in Biosafety cabinets  Standards  Hood , Laminar flow & Bio safety Cabinets  Classification –Bio safety Cabinets TopicsTopics
  • 4.  Bio Safety Level (BSL)-I: Well-characterized agents not known to consistently cause disease in healthy adult humans, and of minimal potential hazard to laboratory personnel and the Environment. Standard practices required:  frequent hand washing  door that can be kept closed when working  limits on access to the lab space when working  no eating, drinking, storage of food in laboratory  care to minimize splashes and actions that may create aerosols (tiny droplets)  decontamination of work surfaces after every use Primary Safety Barriers- Not Required Biosafety LevelsBiosafety Levels
  • 5.  Bio Safety Level (BSL)-II: • Suitable for work involving agents of moderate potential hazard to personnel and the environment. • Generally associated with human disease Safety Equipment (Primary barriers): • Class I or II Bio safety Cabinets or other physical containment devices (eg…. Aerosol tight lid in Centrifuges) are used • Personnel Protection Equipment: Laboratory coats, gloves, face protection as needed Facilities (Secondary barriers): • Provide lockable doors for facilities that house restricted agents • Locating new laboratories away from public areas. • Furniture used in laboratory work should be covered with a non-fabric material that can be easily be decontaminated. • An eye wash station is readily available. Biosafety LevelsBiosafety Levels
  • 6.  Bio Safety Level (BSL)-III: Applicable where work is done with indigenous or exotic agents which may cause serious or potentially lethal disease as a result of exposure by the inhalation route. Safety Equipment (Primary barriers): • Class I or II Bio safety Cabinets • Personnel Protection Equipment: Laboratory coats; gloves; face protection as needed • Facilities (Secondary barriers): • The laboratory has special engineering and design feature . The exhaust air from the laboratory room is discharged to the outdoors • The ventilation to the laboratory is balanced to provide directional airflow into the room Biosafety LevelsBiosafety Levels
  • 7.  Bio Safety Level (BSL)-IV: Dangerous/exotic agents which pose high risk of life-threatening disease, aerosol- transmitted lab infections or related agents with unknown risk of transmission. Safety Equipment (Primary barriers): All procedures conducted in Class III Bio safety Cabinets or Class I or II Bio Safety Cabinets in combination with full-body, air-supplied, positive pressure personnel suit. Facilities (Secondary barriers): BSL-3 plus: • Separate building or isolated zone • Dedicated supply and exhaust and decontamination systems Biosafety LevelsBiosafety Levels
  • 8. Air Filtration in Biosafety cabinetsAir Filtration in Biosafety cabinets
  • 9. •High efficiency particulate air (HEPA) filters •Ultra low particulate air (ULPA) filters Air Filtration in Biosafety cabinetsAir Filtration in Biosafety cabinets Variations in HEPA filter efficiency from 99.95 % to 99.99 % are usually due to manufacturing techniques. Depending upon its quality, a HEPA filter is able to trap from 9,995 to 9,999 of every 10,000 particles An ULPA filter is able to trap from 99,995 to 99,999 of every 100,000 particles ULPA - this is at least 10 times better than HEPA filtration !!!
  • 10. International Standards on Biosafety CabinetInternational Standards on Biosafety Cabinet
  • 11. Why a standard ?Why a standard ?
  • 12. International Standards on Biosafety CabinetInternational Standards on Biosafety Cabinet International Standards are mainly issued by two Organisations: International Standard Organisation (ISO), based in Geneva (Switzerland), Comitee Europeen de Normalisation (CEN) based in Brussels (Belgium). These standards replace all other local standards on the same subject. The aim of the standards is to construct a mechanism to provide manufacturer and user of a common site to specify technical specifications and features. Both construction and testing of the cabinet are covered
  • 13. International Standards on Biosafety CabinetInternational Standards on Biosafety Cabinet Standards: BS-5726 (U.K.) NF X44-201 (France) DIN 12950 (Germany) Now Replaced by EN 12469 EN 12469 European Standards on Design, Construction & Testing of Bio safety Cabinets In addition to comply DIN 12980 for safety handling of cytotoxic drugs NSF-49 (National Sanitation Foundation) In the USA the NSF-49, was one of the first standards on this subject and was the reference point for a lot of manufacturers. NSF is an independent organisation that works as neutral agency to help on the problems that could affect to public health and environment protection. Was published in 1976.
  • 14. Difference between Hood and Safety CabinetDifference between Hood and Safety Cabinet Exhaust of vapors and aerosols from Laboratory to outside No validatable product or personnel protection Validatable personnel, product & environmental protection Filters and blower(s) incorporated HoodHood Safety CabinetSafety Cabinet
  • 15. A fume hood is a ventilation device that is designed to limit exposure to hazardous fumes vapors or dust. A fume hood is typically a large piece of equipment enclosing five sides of a work area, the bottom of which is most commonly located at a standing work height. Two main types exist, ducted and recalculating. The principle is the same for both types: air is drawn in from the front (open) side of the cabinet, and either expelled outside the building or made safe through filtration and fed back into the room. Fume HoodFume Hood Fume Hood
  • 16. Only for Product Protection No Personnel or Environment Protection Laminar FlowLaminar Flow A. front opening B. sash C. supply HEPA filter D. blower Vertical Laminar Flow
  • 17. Laminar FlowLaminar Flow A. front opening B. supply grille C. supply HEPA filter D. supply plenum E. blower F. Grille Horizontal Laminar Flow
  • 18. Classification of Bio Safety CabinetClassification of Bio Safety Cabinet Biological Safety Cabinet Class I Class II Class III Type A Type B Type A I Type A II Type B I Type B II
  • 19. Classification of Biosafety CabinetClassification of Biosafety Cabinet Class I: Personnel and Environment protection. Product is not protected. Class II: Personnel, Product and Environment are protected. Class III: Maximum protection to Personnel, Product and Environment.
  • 20. Classification of Biosafety CabinetClassification of Biosafety Cabinet A. Front opening B. Sash C. Exhaust HEPA Filter D. Exhaust plenum Class I Biosafety Cabinets  This is a negative-pressure, ventilated cabinet usually operated with an open front.  All of the air from the cabinet is exhausted through a HEPA filter either into the laboratory or to the outside.  The Class I BSC is designed for general microbiological research with low- and moderate-risk agents.
  • 21. Classification of Biosafety CabinetClassification of Biosafety Cabinet A. front opening B. sash C. exhaust HEPA filter D. rear plenum E. supply HEPA filter F. blower Class II Type A-I Biosafety Cabinets  It is designed with inward air flow to protect personnel, Product & Environment  HEPA-filtered downward vertical laminar airflow for product protection, and HEPA- filtered exhaust air for environmental protection
  • 22. Classification of Biosafety CabinetClassification of Biosafety Cabinet  Face Velocity 75 fpm  70% re circulated through HEPA Filter  30% Exhausted Through HEPA Filter  Cannot be used for Radio Nucleotides and Toxic Chemicals  Biosafety Levels 2 & 3  User, Product, & Environment Protection Class II Type A-I Biosafety Cabinets
  • 23. Classification of Biosafety CabinetClassification of Biosafety Cabinet Class II Type A-II Biosafety Cabinets A. front opening B. sash C. exhaust HEPA filter D. supply HEPA filter E. positive pressure plenum F. negative pressure plenum Note: The cabinet exhaust needs to be connected to the building exhaust system.
  • 24. Classification of Biosafety CabinetClassification of Biosafety Cabinet Class II Type A-II Biosafety Cabinets  Downward Velocity 100 fpm  70% re circulated through HEPA  Filter30% Exhausted Through HEPA Filter  Can be used for Radio Nucleotides and Toxic Chemicals  Biosafety Levels 2 & 3User, Product, & Environment Protection  Under Negative Pressure to room  Exhaust Air is Hard Ducted
  • 25. Classification of Biosafety CabinetClassification of Biosafety Cabinet Class II Type B-I Biosafety Cabinets A. front opening B. Sash C. exhaust HEPA filter D. supply plenum E. supply HEPA filter F. Blower G. negative pressure exhaust plenum Note: The cabinet exhaust needs to be connected to the building exhaust system.
  • 26. Classification of Biosafety CabinetClassification of Biosafety Cabinet Class II Type B-I Biosafety Cabinets  Downward Velocity 100 fpm  30% re circulated through HEPA Filter  70% Exhausted Through HEPA Filter  Can be used for Radio Nucleotides and Toxic Chemicals (Low Levels / Volatility)  Biosafety Levels 2 & 3User, Product, & Environment Protection
  • 27. Classification of Biosafety CabinetClassification of Biosafety Cabinet Class II Type B-II Biosafety Cabinets A. front opening B. Sash C. exhaust HEPA filter D. supply HEPA filter E. negative pressure exhaust plenum F. supply blower G. filter screen Note: The carbon filter in the building exhaust is not shown. The cabinet exhaust needs to be connected to the building exhaust system
  • 28. Classification of Biosafety CabinetClassification of Biosafety Cabinet Class II Type B-II Biosafety Cabinets  Downward Velocity 100 fpm  No Re-circulation100% Exhausted Through HEPA Filter  Can be used for Radio Nucleotides and Toxic Chemicals  Biosafety Levels 2 & 3User, Product, & Environment Protection  Exhaust HEPA Filtered  Exhaust Hard Ducted out.
  • 29. Classification of Biosafety CabinetClassification of Biosafety Cabinet Class III Biosafety Cabinets A. glove ports with O-ring for attaching arm-length gloves to cabinet B. Sash C. exhaust HEPA filter D. supply HEPA filter E. double-ended autoclave or pass- through box Note: A chemical dunk tank may be installed which would be located beneath the work surface of the BSC with access from above. The cabinet exhaust needs to be connected to the building exhaust system.
  • 30. Classification of Biosafety CabinetClassification of Biosafety Cabinet Class III Biosafety Cabinets  This is a totally enclosed, ventilated cabinet  Gas-tight construction  Offers the highest degree of personnel and environmental protection from infectious aerosols as well as protection of research materials from microbiological contaminants.
  • 31. Cytotoxic Drug Safety CabinetsCytotoxic Drug Safety Cabinets Hepa filter exhaust #2 Hepa filter (pre-filter) #1 Laminar flow filter Carbon filter exhaust #3 (option) Dedicated exhaust flow blower All contaminated areas under NEGATIVE pressure Dedicated Laminar flow blower 70% re circulated HEPA filtered laminar flow Provides sample protection 30% inflow provides operator Safety barrier
  • 32. Can you see the problems ?Can you see the problems ? Window Air vent Ceiling clearance or wall clearance Front Venting Exhaust BEWAR E
  • 33. Location In LabLocation In Lab Location A, E and F are well selected. As well influencing air flows through the Laboratory personnel traffic is avoided. Location B is often influenced from air flows in the Room. Location C is influenced from personnel traffic and Room air flows. Location D is also very often influenced from personnel traffic and air flows. Safety Cabinets should only be placed in quiet areas! Only validated tests can show placement in the laboratory is correct. In this instance the tests are invaluable!