Biological Safety Cabinets. Presented by Mr. Rene Soetens President

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1 Biological Safety Cabinets Presented by Mr. Rene Soetens President

2 Biological Safety Cabinets are referred to by several different names, some of the more common are: BSC, Biosafety Cabinet, Bio-Hood and Class II Laminar Flow Hood Class Biosafety II Laminar Bio-Hood BSCabinet Flow Hood Unfortunately these cabinets are also commonly called and labeled Fume Hoods PROCEDURES INVOLVING THE USE OF HAZARDOUS GASES AND VAPOURS WHICH REQUIRE THE USE OF A FUME HOOD ARE SPECIFICALLY PROHIBITED IN BIOLOGICAL SAFETY CABINETS

3 NSF/ANSI NSF International Standard /ANSI American National Standard Biosafety Cabinetry: Design, Construction, Performance, and Field Certification This Standard, usually referred to as NSF 49, provides the following definitions for Biological Safety Cabinets.

4 NSF/ANSI DEFINITIONS class I: A Class I BSC provides personnel and environmental protection without product protection. Personnel protection is provided as a minimum velocity of 75 lfpm (0.38 m/s) of unfiltered room air is drawn through the front opening and across the work surface. The air is then passed through a HEPA/ULPA filter in the exhaust plenum, providing environmental protection.

5 Class I: NSF/ANSI DEFINITIONS

6 Class I:

7 NSF/ANSI DEFINITIONS class II: Class II (Type A1, A2, and C1 BSCs) BSCs are partial barrier systems that rely on the movement of air to provide personnel, environmental, and product protection. Personnel and product protection is provided by the combination of inward and downward airflow captured by the front grille of the cabinet. Side-to-side cross-contamination of product is minimized by the internal downward flow of HEPA/ULPAfiltered air moving towards the work surface and then drawn into the front and rear intake grills. Environmental protection is provided when cabinet exhaust air is passed through a HEPA/ULPA filter. When used as designed, the cabinet exhaust air may be recirculated to the laboratory or discharged from the building via a canopy connection (Type A1, A2, and C1 BSCs). Exhaust air from Types B1 and B2 BSCs must be discharged to the outdoors via a sealed connection. All Class II cabinets are designed for work involving procedures assigned to biosafety levels 1, 2 and 3. Class II BSCs may be used with procedures requiring BSL-4 containment if used in a BSL-4 laboratory by a worker wearing a positive pressure protective suit. Class II BSCs provide the microbe-free work environment necessary for cell culture propagation and also may be used for the formulation of non-volatile antineoplastic or chemotherapeutic drugs.

8 NSF/ANSI DEFINITIONS class II type A1 cabinets (formerly designated type A): cabinets that maintain minimum average inflow velocity of 75 ft/min (0.38 m/s) through the work access opening; have HEPA/ULPA filtered downflow air that is a portion of the mixed downflow and inflow air from a common plenum (i.e., a plenum from which a portion of the air is exhausted from the cabinet and the remainder supplied to the work area); may exhaust HEPA/ULPA filtered air back into the laboratory or to the environment through an external exhaust system connected to the cabinet with a canopy connection; and have all biologically contaminated ducts and plenums under negative pressure or surrounded by negative pressure ducts and plenums. If working with volatile chemicals, the unit must be connected to an external exhaust system. Type A1 cabinets may be used for work with volatile chemicals if permitted by a chemical risk assessment. NOTE Type A1 BSCs manufactured prior to 2010 are not suitable for work with volatile chemicals due to the contaminated positive pressured plenums that are not surrounded by negative pressure plenums.

9 NSF/ANSI DEFINITIONS class II, type A2 cabinets (when exhausted to the environment were formerly designated type B3): cabinets that maintain a minimum average inflow velocity of 100 ft/min (0.51 m/s) through the work access opening; have HEPA/ULPA filtered downflow air that is a portion of the mixed downflow and inflow air from a common exhaust plenum; may exhaust HEPA/ULPA filtered air back into the laboratory or to the environment through an external exhaust system connected to the cabinet with a canopy connection; and have all biologically contaminated ducts and plenums under negative pressure or surrounded by negative pressure ducts and plenums. If working with volatile chemicals, the unit must be connected to an external exhaust system. Type A2 cabinets may be used for work with volatile chemicals if permitted by a chemical risk assessment.

10 Class II type A1 and A2:

11 Class II type A1 and A2: Negative pressure plenum Positive pressure plenum

12 Class II type A1 and A2:

13 Class II type A1 and A2: NSF/ANSI DEFINITIONS

14 Class II type A1 and A2: Exhaust Canopy

15 NSF/ANSI DEFINITIONS class II type B1 cabinets: cabinets that maintain a minimum average inflow velocity of 100 ft/min (0.51 m/s) through the work access opening; have HEPA/ULPA filtered downflow air composed largely of uncontaminated recirculated inflow air; exhaust contaminated downflow air from a region of the work area via an internal dedicated exhaust plenum and through HEPA/ULPA filter(s) to an external exhaust system with a direct connection and exhausted to the atmosphere; and recirculate the balance of the downflow and inflow air through a supply HEPA/ULPA filter(s); and have all biologically contaminated ducts and plenums under negative pressure or surrounded by negative pressure ducts and plenums. Type B1 cabinets may be used for work with volatile chemicals if permitted by a chemical risk assessment

16 Class II type B1: Either or both of these supply filters may be present in these cabinets

17 Class II type B1:

18 Class II type B1:

19 Class II type B1:

20 Class II type B1:

21 NSF/ANSI DEFINITIONS class II, type B2 cabinets: cabinets that maintain a minimum average inflow velocity of 100 ft/min (0.51 m/s) through the work access opening; have HEPA/ULPA filtered downflow air drawn from the laboratory or the outside air (i.e., downflow air is not recirculated from the cabinet exhaust air); exhaust all inflow and downflow air to the atmosphere through an external exhaust system connected to the cabinet with a direct connection after filtration through a HEPA/ULPA filter without recirculation in the cabinet or return to the laboratory; and have all contaminated ducts and plenums under negative pressure or surrounded by directly exhausted (non-recirculated through the work area) negative pressure ducts and plenums. Type B2 cabinets may be used for work with volatile chemicals if permitted by a chemical risk assessment

22 Class II type B2

23 Class II type B2

24 Class II type B2

25 Class II type B2

26 class II type C1 BSCs: NSF/ANSI DEFINITIONS maintain a minimum average inflow velocity of 100 ft/min (0.51 m/s) through the work access opening; have HEPA/ULPA filtered downflow air composed largely of uncontaminated recirculated inflow air; exhaust contaminated downflow air from a region of the work area via an internal dedicated exhaust plenum and blower, and then through HEPA/ULPA filter(s); recirculate the balance of the downflow and inflow air through a supply HEPA/ULPA filter(s); have all biologically contaminated ducts and plenums under negative pressure or surrounded by negative pressure ducts and plenums; and may exhaust HEPA/ULPA filtered air either back into the laboratory or via a canopy connection to an external system that exhausts to the atmosphere. If working with volatile chemicals, the unit must be connected to an external exhaust system. Type C1 cabinets may be used for work with volatile chemicals if permitted by a chemical risk assessment.

27 Class II type C1

28 Class II type C1 Exhaust airflow

29 Class II type C1 Recirculation/intake airflow

30 Class II type C1

31 Class II type C1 Downflow/recirculation plenum Work Work surface surface airflow Exhaust plenum

32 NSF/ANSI DEFINITIONS

33 Class III: NSF/ANSI DEFINITIONS

34 Class I Biological Safety Cabinet: BSC CLASS TYPE RECAP These cabinets provide a degree of personnel and environmental protection only Air flow is directed away from the researcher, but is not HEPA/ULPA filtered, therefore there is no product protection Similar to a fume hood with a HEPA/ULPA filter on the exhaust system to protect against the release of biohazards Inward air flow ranges from feet per minute (fpm) Can be used with radioisotopes and some toxic chemicals Class II Biological Safety Cabinet: These cabinets provide a degree of personnel, product and environmental protection There are supply air and exhaust air HEPA/ULPA filters Class II A1 cabinets have a minimum inward air flow of 75 fpm and recirculate 70% of the air; Class II A 2, Class II B 1 and Class II B 2 cabinets have a minimum inward air flow of 100 fpm and exhaust either 30% (type A), 70% (type B1) or 100% (type B2) Class II C 1 Cabinets have a minimum inward air flow of 100 fpm and exhaust a portion of the downflow air from a region of the work area and recirculates the balance of the downflow and inflow air Class III Biological Safety Cabinet: These cabinets provide total personnel, product and environmental protection They are hermetically sealed and all procedures are conducted through arm-length rubber gloves Used in the highest level containment labs There are either two HEPA/ULPA filters or HEPA/ULPA filtration and incineration on the exhaust system

35 Biological Safety Cabinet Certification

36 Field Certification Tests Mandatory Tests Tests directly related to containment (personnel and environmental protection) and product protection. Worker Comfort and Safety Tests Tests related to worker comfort and safety and are performed at the request of the customer or the discretion of the certifier. Must be performed to meet NSF/ANSI 49 field certification. These tests are not mandatory for certification

37 Mandatory Tests Downflow velocity profile test: Uniform downflow cabinets Non-uniform (zoned) cabinets Inflow velocity (face velocity) test: Direct inflow measurement method Alternate Inflow measurement methods Exhaust velocity Constricted access opening

38 Mandatory Tests Airflow smoke patterns test: Downflow test View screen retention test Work opening edge retention test Sash/window seal test

39 Mandatory Tests HEPA/ULPA filter leak test Downflow HEPA/ULPA filter Exhaust HEPA/ULPA filter Scan test Total leakage test when filter cannot be scanned (ducted exhaust)

40 Mandatory Tests Cabinet integrity test (Only mandatory for positive pressure plenum cabinets) Pressure decay test Soap bubble test

41 Mandatory Tests Site installation assessment tests: Alarm functions Airflow alarms (if present) Sash alarms Blower interlocks (on B2 cabinets) Exhaust system performance Exhaust Canopy (smoke test) must be under negative pressure with the cabinet operating.

42 Worker Comfort and Safety Tests Lighting intensity test Vibration test Noise level test Electrical leakage and ground circuit resistance and polarity tests (Newer cabinets are required to comply with UL , which if requested should be performed by a licensed electrician)

43 How Often do You Need to Certify? NSF/ANSI Annex F

44 Airflow Specifications CETA Specification Guide A comprehensive list of specifications in one document Manufacturer approved NSF Blue Book As Tested at NSF Manufacturer As tested by individual manufacturers

45 Airflow Specifications Downflow Inflow Exhaust There can be many airflow specifications for a single model of cabinet CFM For example the following velocity and volume specifications are all for one model of cabinet F , C 60-70, B CFM F , C 60-70, B F , C 62-72, B F , C 62-72, B F 66-76, C 80-90, B F 79-89, C 88-98, B

46 Biological Safety Cabinet Repair Biological Safety Cabinet Repair

47 Biological Safety Cabinet Repair In most cases repairs to a Biological Safety Cabinet should only be performed by a qualified service technician With the exception of externally mounted fluorescent lighting, repair or adjustment of electrical components located outside of the containment envelope of the BSC may affect containment HEPA filters cannot be cleaned and replacement of these filters or repairs to internal components located within the containment envelope of the BSC should only be performed after gaseous decontamination.

48 Biological Safety Cabinet Repair Formaldehyde Gas Decontamination The most common type of biological decontamination procedure uses solid paraformaldehyde which is heated to produce formaldehyde gas in high concentration The gas must remain in contact with the contaminated areas of the BSC for several hours to ensure complete inactivation of any viable organisms Decontamination. decontamination.

49 Biological Safety Cabinet Repair Formaldehyde Gas Decontamination Formaldehyde Gas is considered effective on most biological agents and is widely accepted as for biological decontamination

50 Biological Safety Cabinet Repair Formaldehyde Gas Decontamination Concerns Prions and slow viruses Formaldehyde gas is considered to be ineffective on these agents Chemotherapy hoods Chemotherapy agents are chemicals and cannot be rendered safe by exposure to formaldehyde gas Safety Standards Pesticide regulations Inconsistent performance in the field

51 Biological Safety Cabinet Repair Decontamination Formaldehyde Gas Chlorine Dioxide Vaporous Hydrogen Peroxide Some of these methods have significant disadvantages which can render them impractical as a standard decontamination procedure

52 The Care and Feeding of your Biological Safety Cabinet

53 The Ten Commandments 1. Preparation Turn off U.V. Lamp UV lighting is not recommended in Class II biosafety cabinetry per NSF/ANSI p.20 section Check air grills for obstructions Turn on cabinet blower Allow air to purge the workspace > 5minutes

54 The Ten Commandments 2. Disinfection Spray or swab all interior surfaces with appropriate disinfectant Allow to air dry

55 The Ten Commandments 3. Assemble Material Introduce only materials required to perform procedure Place material such that clean and contaminated items are not in contact Place contaminated materials to one side at rear (not applicable to Class II type C1 cabinets) Ensure view screen is properly located and secure

56 The Ten Commandments 4. Pre-Use Purge Allow air purge period of > 5 minutes with no activity inside

57 The Ten Commandments 5. Pre-Use Personal Procedures Don Protective clothing, gloves, face mask etc. as appropriate

58 The Ten Commandments 6. Perform Procedures Introduce hands into work space, work carefully and methodically (i.e. from clean to work area to discard) Move slowly to avoid disruption of containment air barrier (air inflow at access opening) DO NOT remove your hands from the work space until procedures are complete and all critical and contaminated material is secured Remove gloves and place into contaminated waste container

59 The Ten Commandments 7. Post-Use Purge Allow air purge period of > 5 minutes with no activity inside

60 The Ten Commandments 8. Post-Use Personal Procedures Remove Protective clothing, face mask etc. and dispose of as appropriate

61 The Ten Commandments 9. Terminal Disinfection Don gloves, remove materials to incubator etc., to biohazard container, autoclave as appropriate Spray or swab all interior surfaces with an appropriate disinfectant

62 The Ten Commandments 10. Shutdown Turn off Fluorescent lamp Turn off cabinet blower

63 Your Biological Safety Cabinet Service Provider is

64 What assurance of quality services does Con-Test provide? Experience In House Training Program Technical Support Latest Test Equipment Documentation Control Adequate Insurance Coverage Customer Support ISO 9001:2008 Company NSF Accredited Technicians NEBB Accredited Member of: ABSA ASHRAE CETA IEST PDA

65 QUESTIONS?

66 THANK YOU FOR ATTENDING

67

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