US Environmental Protection Agency
Office of Pesticide Programs
Office of Pesticide Programs
Microbiology Laboratory
Environmental Science Center, Ft. Meade, MD
Standard Operating Procedure for
Use and Maintenance of Biological Safety Cabinets
SOP Number: EQ-11-01
Date Revised: 05-11-17
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SOP No. EQ-11-01
Date Revised 05-11-17
Page 1 of 7
SOP Number
EQ-11-01
Title
Use and Maintenance of Biological Safety Cabinets
Scope
The purpose of this SOP is to describe the procedures for the use and
maintenance of the Biological Safety Cabinets (BSC).
Application
Proper use and cleaning of the BSCs are the responsibilities of
laboratory personnel. Performing maintenance and repairs on the
BSCs and maintaining annual certification of the BSCs are the
responsibilities of the Facility Manager or his designees (e.g.,
operations, maintenance and repair contractors).
Approval Date
SOP Developer:
Print Name:
SOP Reviewer
Print Name:
Quality Assurance Unit
Print Name:
Branch Chief
Print Name:
Date SOP issued:
Controlled copy
number:
Date SOP withdrawn:
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SOP No. EQ-11-01
Date Revised 05-11-17
Page 2 of 7
TABLE OF CONTENTS
Contents Page Number
1.
DEFINITIONS
3
2.
HEALTH AND SAFETY
3
3.
PERSONNEL QUALIFICATIONS AND TRAINING
3
4.
INSTRUMENT CALIBRATION
3
5.
SAMPLE HANDLING AND STORAGE
3
6.
QUALITY CONTROL
3
7.
INTERFERENCES
3
8. NON-CONFORMING DATA
4
9.
DATA MANAGEMENT
4
10.
CAUTIONS
4
11.
SPECIAL APPARATUS AND MATERIALS
4
12.
PROCEDURE AND ANALYSIS
4
13.
DATA ANALYSIS/CALCULATIONS
6
14.
FORMS AND DATA SHEETS
6
15.
REFERENCES
7
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SOP No. EQ-11-01
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1. Definitions
Abbreviations/definitions are provided in the text.
2. Health and
Safety
Follow procedures specified in SOP MB-01, Laboratory Biosafety. The Study
Director and/or lead analyst should consult the Safety Data Sheet for specific
hazards associated with products.
3. Personnel
Qualifications
and Training
Refer to SOP ADM-04, OPP Microbiology Laboratory Training.
4. Instrument
Calibration
1. BSC working parameters are controlled by the Facility Manager or his
designee.
2. Work zone air downflow is uniform and held to an average velocity of 60
linear feet per minute (LFPM) - 0.30 m/s measured in the plane of the
bottom edge of the viewing window. The BSC will alarm if the average
velocity falls below 54 LFPM or above 64 LFPM. The significance of
this airflow control is that the work zone air is flowing down like a huge
piston carrying contaminants rapidly out of the work zone with 24 air
changes per minute. The work zone airflow is ducted such that the
quantity of air leaving through the rear perforated area is half of the
downflow air quantity. The remainder, that is the other half of the
downflow air, and the air entering through the work access opening flow
into the front perforated area. Room air flowing into the 8 inch (203 mm)
work access opening forms the air barrier protecting personnel at 105 ±5
feet per minute (FPM) - 0.53 ±0.25 m/s. The exhaust volume is displayed
on the front panel. The BSC alarms if the exhaust volume falls below 980
cubic feet per minute (CFM) or rises to above 1250 CFM.
5. Sample
Handling and
Storage
Not applicable
6. Quality Control
1. For quality control purposes, the required information is documented on
the appropriate form(s) (see section 14).
7. Interferences
1. If a procedure requires the use of a flame (e.g., to flame sterilize hooks,
etc.), place a burner with a pilot light in the center of the work surface, so
that the flame can be ignited only when required. Do not use constant
flame gas burners.
2. Gas burners should only be used from the center of the work surface to the
right rear, where resulting air turbulence will have a minimal effect. Do
not use gas burners on the left of the work surface due to its influence on
the electronic airflow control system (see section 15).
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SOP No. EQ-11-01
Date Revised 05-11-17
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3. A permanent paper catch is installed behind the rear divider panel of the
work zone. This area forms the return air path to the motor/blower. If the
air flow is blocked, it could seriously affect the performance of the
cabinet. Regularly verify that the paper catch is clear of any paper
products that may disrupt the air flow.
8. Non-
conforming
Data
1. Management of non-conforming data will be consistent with SOP ADM-
07, Non-Conformance Reports.
9. Data
Management
1. Data will be archived consistent with SOP ADM-03, Records and
Archives.
10. Cautions
1. Failure to clean the ultraviolet lamps in the BSC will reduce the lamps'
effectiveness.
2. Check grills and diffuser grids periodically for spilled or splashed
nutrients which may cause contamination of the work surface.
3. Do not use halogenated materials (e.g., bleach). These materials are not
recommended for use on stainless steel surfaces of the BSC.
4. Exercise care to ensure that no items are placed over the front intake grills.
5. Avoid passing contaminated materials over uninoculated cultures or clean
glassware to avoid contamination.
6. Do not use cabinet as a repository for excess lab equipment during periods
of non-operation.
11. Special
Apparatus and
Materials
The list of BSCs is maintained as an attachment to the SOP. See section 14.
12. Procedure and
Analysis
Follow the instructions below for operation/use/maintenance of the BSC.
12.1 Use and
Operation
a. Laboratory staff performing work in the BSC must wear a long-
sleeved lab coat and gloves even when not manipulating culture. This
minimizes the shedding of skin flora into the work area and
concurrently protects the hands and arms from viable agent
contamination (see section 15).
b. The laminar flow BSCs create an environment that has little to no
airborne particulates. It is not necessary to flame tube and flask
necks.
c. Turn on the blowers, lights, and outlets, and allow equipment to
operate for a minimum of 15 minutes.
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SOP No. EQ-11-01
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d. Disinfect the interior surface of the BSC work space thoroughly by
spraying/wetting with an appropriate amount of an EPA registered
hospital disinfectant; allow to stand for the label-specified contact
time before wiping.
e. Place apparatus and materials into the cabinet. Arrange the work area
so that clean and dirty (used) materials are well separated.
f. Transfer viable materials (such as test cultures) as deeply into the
cabinet (away from open face) as possible.
g. Allow purging of the work space without any user activity for 2-3
minutes after materials and apparatus have been placed in the unit(s).
h. Complete the BSC Monitoring Record Form (see section 14)
immediately prior to use. Check the appropriate box ("Yes" or "No")
to indicate if the unit's Downflow (FPM) and Exhaust (CFM) are
within the acceptable range (see section 4.2).
i. After completion of work, allow the BSC to run for 2-3 minutes
without activity to purge the unit.
j. Remove all materials, cultures, and equipment and decontaminate
BSC as in section 12.2.
k. When working with bacteria in spore form (e.g., Bacillus subtilis
spore suspensions, Clostridium difficile spore suspension, spore strips,
etc.), turn on the ultraviolet light and leave it on overnight.
1. Refer to the laboratory's Biosafety Plan for Bacillus anthracis for
BSC clean up procedures following work with select agents.
m. Turn off lights and outlets. Turn off blowers as per facility
recommendations.
n. Record all maintenance activities, incidents or corrective actions taken
on the BSC Monitoring Record Form (see section 14).
12.2 Quarterly
Maintenance:
a. Thoroughly clean and disinfect all BSCs on a quarterly basis.
b. Remove panel and grill from the BSC and set aside.
c. Disinfect all surfaces and the panel grill of the BSC thoroughly by
spraying/wetting with an appropriate amount of a hospital disinfectant
and allow to stand for the label-specified contact time before wiping.
d. Replace the panel and grill, and properly secure.
e. Clean the ultraviolet lamps in the BSC with a lint-free cloth dampened
with alcohol.
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SOP No. EQ-11-01
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f. Record quarterly maintenance on the BSC Monitoring Record Form
(section 14).
12.3 Service and
Certification
a. Notify the Facilities Helpline at (410) 305-4357, if the BSC needs
service. The Facility Manager or his designees investigate and make
a service call if necessary. Laboratory staff notes the event on the
BSC Monitoring Record Form (see section 14) and documents any
corrective action taken.
b. Certification: The BSCs are certified annually by a qualified
technician. A series of tests are performed on the BSC during
recertification including the HEPA filter leak test and a measurement
of the downflow blower speed (designated "Blower" or "Blower
Speed" on the certificate and report).
c. Scheduling for recertification: Scheduling recertification is the
responsibility of the Facility Manager or his designee. Certificates are
maintained by the Facility Manager. The OPP Microbiology
Laboratory requests a copy of the Test and Certification Reports each
year for review.
d. HEPA filter usage and replacement:
i. HEPA filters on each BSC do not need to be replaced under
normal usage, and barring an accident (puncture), until the
exhaust volume or the access flow velocity cannot be
maintained. Making this determination and scheduling HEPA
filter replacement is the responsibility of the Facility Manager or
his designee.
ii. The Facility Manager takes into consideration the downflow
blower speed (if >90%) and other indicators (e.g., age - 8 to 10
years) to determine the need for replacement of the HEPA filter.
iii. Once the HEPA filter is replaced, the BSC is recertified and the
certification is annotated by a Performance Certification sticker.
13. Data Analysis/
Calculations
None
14. Forms and Data
Sheets
Forms are stored separately from the SOP under the following file names:
Attachment: Current Inventory of BSCs
BSC Monitoring Record Form EQ-11-01 F1.docx
BSC Quarterly Maintenance Record Form EQ-11-01 F2.docx
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SOP No. EQ-11-01
Date Revised 05-11-17
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15. References
1. Bordner, R.H., Winter, J.A. &, Scarpino, P.V., eds. 1978.
Microbiological Methods for Monitoring the Environment, Water and
Wastes. EPA 600/8-78-017, Environmental Monitoring & Support Lab.,
U.S. Environmental Protection Agency, Cincinnati, Ohio.
2. NuAire Labguard Class II, Type B2 Laminar Flow Biological Safety
Cabinet Operation and Maintenance Manual.
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