working safely in your biological safety cabinet
DESCRIPTION
Working Safely In Your Biological Safety Cabinet. Training Overview. Part I. HEPA & ULPA Filters Part II.Different Types of Cabinets Class I, Class II and Class III BSC Differences between A2 and B2 BSC International Standards Part III.Biosafety Cabinet Certification - PowerPoint PPT PresentationTRANSCRIPT
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Working Safely In Your Biological Safety Cabinet
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Training Overview
Part I. HEPA & ULPA FiltersPart II. Different Types of Cabinets
Class I, Class II and Class III BSCDifferences between A2 and B2 BSCInternational Standards
Part III. Biosafety Cabinet CertificationPart IV. Common user mistakesPart V. Demonstration on how to use a BSC
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HEPA & ULPA Filter
HEPA: High Efficiency Particulate Air ULPA: Ultra Low Penetration Air
Important definitions:- HEPA: 99.99% - at 0.3 microns - ULPA: 99.999% - at 0.12 microns
Note: The “classical” definition of HEPA filter is 99.97% at 0.3 microns, but nowadays all BSC and LF in US use 99.99% at 0.3 m
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Particle Size Comparison
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HEPA/ULPA CapabilityRemoves a broad range of airborne contaminants:• Fine dust• Smoke• Bacteria (typical size: 500 to 0.3 micron)• Soot• Pollen• Radioactive particles • Impurity ion -> can affect Integrated Circuit speed
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Types of Cabinets
•Biohazard Safety Cabinet (BSC)
•Laminar Flow Cabinet (LFC)
•Fume Hood
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• Product protection (no personnel protection)• Not for biohazard agents or chemical fumes
Laminar Flow Cabinets
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• Class I BSC: Personnel and Environment Protection
• Class II & III BSC: Personnel, Product and Environment Protection
• HEPA filters (not for chemical vapours)
Biosafety Cabinets
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• Removes toxic chemical (ducting sys./ductless)• No HEPA filter -> not for biohazard agents
Fume Hoods
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Class 1 BSC• Only operator protection (not product)
• Biosafety level 1, 2, 3
• Inflow away from operator
• HEPA filtered exhaust to environment
• Current trend: to Class 2
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Class 2 A2
•Recirculating cabinet airflow
•No chemical / toxic vapours containment without ducting
•Approx. 70% recirculating and 30% exhaust
Exhaust Airflow approx. 30%
Room AirDownflow approx.
70%
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Class II Type A2 can’t be used for chemical vapours. Chemical vapour buildup in cabinet & lab is dangerous. Class II Type A2 BSC can be ducted using a non-airtight (thimble) duct.
Class II Type A2 BSC: Ducting
Thimble duct: have holes for room airBldg. exhaust fluctuations affect cabinet airflow
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Class II B2 BSC Airflow
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Class 2 Type B2
•Total exhaust cabinet (no recirculation)•Both biological and chemical vapor containment
•Care with chemical: should not destroy filter•High exhaust air volume•Interlock system: if building exhaust fails•Dedicated exhaust fan with dynamic balancing•Exhaust fan -> precisely match the cabinet: - airflow volume - static pressure•Inflow and downflow are opposite each other•High level of expertise for install & maintenance
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Class 2 Type B2 BSC: Airflow
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Class 3 BSC
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International Standards for Class II
• US Standard ANSI/NSF49
• European Standard EN12469
• Japanese Industrial Standard JIS K3800
• South African Standard SABS VC 8041:2001
• British Standard BS5726*
• German Standard DIN12950 Teil 10*
• French Standard NF X44-201:1984*
*now obsolete. Replaced with the harmonized EN12469
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Biosafety Cabinet Certification
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Biosafety Cabinet Certification
• The functional operation and integrity of each BSC should be certified to national or international performance standards at the time of installation and regularly thereafter by qualified technicians
• Evaluation of the effectiveness of cabinet containment should include tests for cabinet integrity, HEPA filter leaks, inflow/downflow velocity profile, air flow smoke pattern, and alarms and interlocks.
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No Test Field
EN & NSF Production
Per EN Production
Per NSF Type
Per EN Type
Per NSF 1 Inflow velocity √ √ √ √ √ 2 Downflow velocity √ √ √ √ √ 3 HEPA / ULPA filter leak test √ √ √ √ √ 4 Smoke pattern / airflow visualization √ √ √ √ √ 5 Site installation assessment (ex: alarm) √ √ √ √ √ 6 Light intensity √ √ √ √ 7 Noise level √ √ √ √ 8 Vibration level √ √ √ √ 9 Electrical safety testing to IEC 61010 √ √ √ √
10 Pressure retention / soap bubble √ √ √ 11 Microbiological personnel protection √ √ 12 Microbiological product protection √ √ 13 Microbiological cross-contamination √ √ 14 Microbiological performance envelope √ 15 Motor / blower performance √ 16 Drain spillage trough leakage √ 17 Resistance to overturning √ 18 Resistance to distortion √ 19 Resistance to deflection √ 20 Resistance to tipping √ 21 Secondary inflow velocity correlation to DIM √ 22 Powder coating chemical resistance √ 23 Powder coating abrasive resistance √ 24 Cabinet design evaluation √
Testing List
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Common User Mistakes
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Common User Mistakes
• Confusing a vertical laminar flow cabinet for a Class II BSC
• Failure to identify the type of BSC needed for their operations
• Class II B2 cabinet not necessarily “safer” than a Class II A2 cabinet (A2 cabinets can also be ducted out when necessary)
• Inappropriate choice of installation site / cabinet location
• Inappropriate usage / maintenance of the BSC
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Common User Mistakes: Illustration
Blocking of airflow perforations with objects
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Proper Installation / LocationExhaust filter area: Especially susceptible to disruptive air currents. Clearance of 40 cm (minimum) is recommended between the highest point of the cabinet and the ceiling.
Biosafety Cabinet
Barrier AirflowAir Inlet
Return Air Grille Personnel Traffic
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• Slow deliberate movements that will not disrupt airflow, minimize arm movement
• When an alarm is activated, do NOT use the cabinet
• After usage, wipe down the cabinet with cleaning agents
• Work as far into the cabinet as possible• Work starting from clean to “dirty” objects• Do not block airflow perforations with
objects/equipments
Proper Operation
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• Recommended recertification for BSC’s:
– Upon installation at customer’s site
– On-site annually in accordance with international standards
– When the cabinet is moved
Annual Testing
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• Germicidal UV lamps are not substitutes for proper cleaning of BSC workzone
• May cause performance degradation
• May compromise personnel safety when proper precautions are not taken
UV Lamps
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Bunsen Burners• The use of Bunsen burners in LFC and BSC
is discouraged• Compromises cabinet’s operator and cross-
contamination protection when used
Filter damage due to bunsen burner usage within workzone
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Reporting of accident/incident
• If there is a spillage in the BSC, notify the PI(s)/supervisor(s)/laboratory officer(s) immediately and decontaminate all objects in the cabinet without turning off the cabinet.
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Demonstration