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Biosafety Levels, Biological Safety Cabinets and Biosafety Laboratory Constru...Ravi Kant Agrawal
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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!