BIOSAFETY CABINETS

UNIT OF TROPICAL LABORATORY MEDICINE

January 2012
Hilde De Boeck
 Hood for sale

Sihanouk Hospital Centre of Hope, Cambodia

Mycobacterium tuberculosis culture

Baker Laminar Flow Hood
Susan’s Book
 Table 3: Summary of biosafety level requirements

Biosafety Level

Source: WHO (2004), Laboratory Biosafety Manual, 3th Edition

• Include class
microorganisms

Source: WHO (2004), Laboratory Biosafety Manual, 3th Edition

Choosing a cabinet

Types of cabinets:

Personal protection

Product and

Product Protection personal protection

Horizontal and vertical outflow cabinets are not biological safety cabinets!!
Types of cabinets: General

Source: WHO (2004), Laboratory Biosafety Manual, 3th Edition

Types of cabinets: Class I

Class I biological safety cabinet

• Operator protection
• Environment protection
• No product protection
• Exhaust through HEPA (lab,
exhaust, outside
• 0% recirculation, 100% exhaust
• front: min. 0.38m/s velocity

Source: WHO (2004), Laboratory Biosafety Manual, 3th Edition

Types of cabinets: Class ?

Source: WHO (2004), Laboratory Biosafety Manual, 3th Edition

Types of cabinets: Class II

Class II biological safety cabinet

• Operator protection
• Product protection
• Environment protection
• Inflow: min 0.38m/s
• 70% recirculation, 30% exhaust

• types A1, A2, B1, B2

~ air intake velocity
~ amount of air re-circulation
Source: WHO (2004), Laboratory Biosafety Manual, 3th Edition
~ exhaust system (room/outside)
Types of cabinets: Class II

Source: WHO (2004), Laboratory Biosafety Manual, 3th Edition

Types of cabinets: Class III

Class III biological safety cabinet

• Highest level of personnel protection

• Risk 4 agents
• exhaust through 2 HEPA filters
• access work surface: heavy duty
gloves

Source: WHO (2004), Laboratory Biosafety Manual, 3th Edition

Types of cabinets: connections

There are three ways to connect a safety

cabinet to a duct:
• Direct connection
• Direct with a by-pass
• Connection via a thimble

DIRECT CONNECTION
 Types of cabinets: connections

BY-PASS

VIA THIMBLE
Selection of a biological cabinet

In accordance with type of protection:

– Personnel protection
– Product protection
– Risk group 1-4 microorganisms
– Radionuclide – volatile toxic chemicals

but also..
use
maintenance
training
emplacement
Selection of a biological cabinet

Standards specify requirements for cabinet construction

and performance criteria.

International standards:
• European standard EN 12469
• American Standard ANSI/NSF49
• Australian standard AS2252
• Japanese standard JIS K 3800
• ……..
Europe before 2000:
BS5726
DIN12950

European STANDARD
EN12469:2000
 Equipment Life Cylce

Performance needs Emplacement

Acquisition & Implementation &
Facility requirements Verification/calibration
Total cost Selection Installation Documentation
Ease of use Procedures
Purchase conditions Maintenance
Service Training
Consummables

Equipment
Life Cycle

Monitor
Safety issue
Maintenance
(salvaging spare parts)
Troubleshooting
End of Life Use Service/repair
Performance Testing

What should be done?

Why test?
• Safety: workers protection & environment
• Quality assurance Each BSC should be certified to national or
international performance standards
• Legal requirements
When?
- After installation
- Regularly intervals (once a year…)
- After moving

ALWAYS PERFORM DESINFECTION BEFORE MAINTENANCE _ PERFORMANCE

CHECK
Disinfection
Disinfection
Performance Testing

What do we check?
FIRST -visual inspection:

• General condition
• Clean and free working area
• Functionality of displays
• Damaged filters-sealing problems
• Pre-filters
Pre-filters

• Cheap glassfiber filter-material, to protect the expensive

HEPA-filters

• To be changed every year

• Low cost exchange

• Eff. 80 – 90% (EN 779)

• Use doubles lifetime of a HEPA

Pre-filters (if fitted) must be replaced prior to start velocity

recording
Performance Testing

Class II EN 12469 (in practice)

• Smoke test: visualisation air flow patterns
• Down flow velocity: range 0.25-0.5m/s, all measurement +/-20% of mean
• Face – or inflow velocity : > 0,4m/s
• Filter leak testing: HEPA
• Alarms: airflows and window

(Sound level
Vibration
Lightning
Materials)
….
Performance Testing

Flow pattern
=> Smoke testing

Inflow:
inward over the whole area of the front aperture

Down flow:
downwards without turbulence

… But at least your BSC is safe…

Performance Testing

Airflow velocity
Down flow
Measurements have to be taken at
• 8 positions
• 5-10cm above the top edge of the front aperture

Criteria:
- All values have to be >0,25m/s and < 0,5m/s
- According manufacturer’s specifications
- No individual measurements should deviate more than 20% from
mean
Performance Testing

Airflow velocity
Inflow
Measurements need to be taken at
• 9 positions
• In the front aperture
Room condition: door, airco,etc.

Criteria:
Mean inflow velocity > 0,4m/s and
According to manufacturer’s specifications
Performance Testing

Control of HEPA filter efficiency

Option 1
Filter test: Particle counter

Option 2
Efficiency: Photometer + test aerosol

Criteria:
Local penetration <=0,01% AEROSOL:
0 parts • DOP (di-octyl-phalate)
Carcinogenic
• EMERY 3004 (poly-alpha olefin)
Currently in use
HEPA

• HEPA = High Efficiency Particulate Air

• ULPA = Ultra Low Particulate Air
Remove particles >=0,3µm (bacteria, spores, viruses)
with efficiency 99,99% (EN 12469)

100.000
10 Particles
particles
HEPA
HEPA

What in case of leaks??

• Mechanical (pipette tips, broken vials,..)
• Aging (normal life time: 6-10y)
• …..

REPAIR OR REPLACE?

Repair> budget, no/little downtime, no prior bio-decontamination??

PPE! Gloves, mask, Tyvek suite,…

Use and work… General rules

• Keep a log
• Prepare you’re work
• Don’t overfill
• Limit movement inside-out
• Disinfection materials and cabinet before and after
use
• Be prepared for spills..
What to do in case of spills

• Don’t switch off

• Cover with absorbing paper
• Apply disinfectant (alcohols, hypochlorite, dettol,..)
• Allow contact time (30min-2hrs)
• Remove absorbing paper (infectious waste!!)
• Disinfect all items
• Inspect and clean
Voltage and frequency

220–240 V/60 Hz 220–240 V/50 Hz

100–127 V/60 Hz 100–127 V/50 Hz
 REFERENCES: TB + accreditation

http://www.afro.who.int/en/clusters-a-programmes/hss/blood-safety-
laboratories-a-health-technology/blt-highlights/3859-who-guide-for-the-
stepwise-laboratory-improvement-process-towards-accreditation-in-
the-african-region-with-checklist.html

WH0 1998
Lab Services in TB control
Part 1 - 3
GLI, 2013

http://www.stoptb.org/wg/gli/accreditation.asp
References: Biosafety
EQUIPMENT VALIDATION
Installation Qualification
Example URS (User Requirement Specifications)