N S F.
O R G
NSF International Standard
American National Standard
NSF/ANSI 49-2024
Biosafety Cabinetry:
Design, Construction,
Performance, and
Field Certification
NSF International, an independent, not-for-profit,
nongovernmental organization, is dedicated to
being the leading global provider of public health
and safety-based risk management solutions
while serving the interests of all stakeholders.
This standard is subject to revision.
Contact NSF to confirm this revision is current.
Users of this standard may request clarifications and
interpretations, or propose revisions by contacting:
Chair, Joint Committee on Biosafety Cabinetry
c/o NSF International
789 North Dixboro Road, P.O. Box 130140
Ann Arbor, Michigan 48113-0140 U.S.A.
Phone: (734) 769-8010 Email: info@nsf.org
Web: <www.nsf.org>
NSF/ANSI 49 – 2024
NSF International Standard /
American National Standard
for Biosafety Cabinery –
Biosafety Cabinetry: Design, Construction,
Performance, and Field Certification
Standard Developer
NSF International
Designated as an ANSI Standard
February 3, 2024
American National Standards Institute
i
Prepared by
The NSF Joint Committee on Biosafety Cabinetry
Recommended for adoption by
The NSF Public Health Council
Adopted by
NSF International
June 1976
Revised May 1983 Revised June 1987 Revised May 1992
Revised March 2002 Addendum November 2002 Revised February 2004
Revised September 2004 Addendum October 2004 Addendum March 2005
Revised July 2007 Revised October 2008 Revised June 2009
Revised September 2010 Revised November 2010 Revised November 2011
Revised July 2012 Revised February 2015 Revised March 2017
Revised January 2019 Revised March 2020 Revised March 2021
Revised February 2023 Revised November 2024
Published by
NSF International
P.O. Box 130140, Ann Arbor, Michigan 48113-0140, U.S.A.
For ordering copies or for making inquiries with regard to this standard, please reference the designation
“NSF/ANSI 49 – 2024.”
Copyright 2024 NSF International
Previous editions © 2022, 2021, 2020, 2019, 2017, 2015, 2012, 2011, 2010, 2009, 2008, 2007, 2005, 2004,
2002, 1992, 1987, 1983, 1976
Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any
means, electronic or mechanical, including photocopying and microfilm, without permission in writing from
NSF International.
Printed in the United States of America.
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Disclaimers 1
NSF International (NSF), in performing its functions in accordance with its objectives, does not assume or
undertake to discharge any responsibility of the manufacturer or any other party. The opinions and findings
of NSF represent its professional judgment. NSF shall not be responsible to anyone for the use of or reliance
upon this standard by anyone. NSF shall not incur any obligation or liability for damages, including
consequential damages, arising out of or in connection with the use, interpretation of, or reliance upon this
standard. It is the responsibility of the user of this standard to judge the suitability of the ANS for the user’s
purpose.
Participation in NSF standards development activities by regulatory agency representatives (federal, state,
or local) shall not constitute their agency's endorsement of NSF or any of its standards.
Preference is given to the use of performance criteria measurable by examination or testing in
NSF standards development when such performance criteria may reasonably be used in lieu of design,
materials, or construction criteria.
A dual-unit technical standard may use both non-SI (English System of Measurement) and SI units of
measure (SI is the International System of Units, commonly known as the metric system). Unless stated
otherwise, whichever system provided in the primary position is the official and normative unit value. Units
may be provided in parentheses immediately following the primary units when required for clarity or
informational purposes. As such, these values stated are not interchangeable, should be regarded
separately, and used independently. Interchanging values from both unit systems may result in errors.
The illustrations, if provided, are intended to assist in understanding their adjacent standard requirements.
However, the illustrations may not include all requirements for a specific product or unit, nor do they show
the only method of fabricating such arrangements. Such partial drawings shall not be used to justify
improper or incomplete design and construction.
At the time of this publication, examples of programs and processes were provided for general guidance.
This information is given for the convenience of users of this standard and does not constitute an
endorsement by NSF International. Equivalent programs and processes may be used.
Unless otherwise referenced, the annexes are not considered an integral part of NSF standards.
The annexes are provided as general guidelines to the manufacturer, regulatory agency, user, or certifying
organization.
1 The information contained in this disclaimer is not part of this American National Standard (ANS) and has not been
processed in accordance with ANSI’s requirements for an ANS. Therefore, this disclaimer may contain material that
has not been subjected to public review or a consensus process. In addition, it does not contain requirements necessary
for conformance to the standard.
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Abbreviations
The following table is provided as a reference for unit abbreviations for common forms of measurement
used within NSF documents.
second s ampere A
minute min cubic feet per
CFM
hour h minute
A-weighted
time day d dBA
decibel
week wk foot-candle fc
month mo hertz Hz
year yr kilopascal kPa
inch in lux lux
foot ft Pascal Pa
miscellaneous
yard yd parts per million ppm
micrometer µm pounds per
psi
length nanometer nm square inch
millimeter mm pounds per
square inch psig
centimeter cm gauge
meter m radian rad
kilometer km revolutions per
rpm
milliliter mL minute
liter L volts V
liters per day LPD
liters per minute LPM
ounce oz
liquid measure pint pt
quart qt
gallon gal
gallons per
GPM
minute
gallons per day GPD
microgram µg
picogram pg
nanogram ng
milligram mg
centigram cg
weight
gram g
kilogram kg
pound lb
ton t
metric ton mt
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Contents
1 General ................................................................................................................................................... 1
1.1 Scope ............................................................................................................................................ 1
1.2 Minimum requirements ................................................................................................................. 1
1.3 Variations in design and construction ........................................................................................... 1
2 Normative references ............................................................................................................................. 1
3 Definitions ............................................................................................................................................... 3
4 Materials ............................................................................................................................................... 10
4.1 General ....................................................................................................................................... 10
4.2 Interior work surfaces ................................................................................................................. 10
4.3 Exposed interior surfaces ........................................................................................................... 10
4.4 Other interior and exterior surfaces ............................................................................................ 10
4.5 Materials and finishes ................................................................................................................. 10
5 Design and construction ....................................................................................................................... 12
5.1 General ....................................................................................................................................... 12
5.2 Surface decontamination ............................................................................................................ 12
5.3 Gas / vapor decontamination ...................................................................................................... 12
5.4 Canopy exhaust connection ....................................................................................................... 12
5.5 Direct exhaust connection .......................................................................................................... 13
5.6 Duct and plenum design ............................................................................................................. 13
5.7 Internal corners and angles ........................................................................................................ 13
5.8 External corners and angles ....................................................................................................... 13
5.9 Joints and seams ........................................................................................................................ 13
5.10 Fastening methods ..................................................................................................................... 14
5.11 Removable panels ...................................................................................................................... 14
5.12 Stability ....................................................................................................................................... 14
5.13 Provision for mounting ................................................................................................................ 14
5.14 Legs and feet .............................................................................................................................. 14
5.15 Reinforcing and framing .............................................................................................................. 14
5.16 Doors and covers ........................................................................................................................ 14
5.17 Louvers and openings ................................................................................................................ 15
5.18 Tracks and guides....................................................................................................................... 15
5.19 Filters .......................................................................................................................................... 15
5.20 Gaskets and sealants ................................................................................................................. 16
5.21 Stopcocks and service outlets .................................................................................................... 16
5.22 Alarms ......................................................................................................................................... 17
5.23 Electrical components ................................................................................................................. 18
5.24 Lighting ....................................................................................................................................... 18
5.25 Gauges, pressure transducers, and switches ............................................................................ 19
5.26 Drain spillage trough ................................................................................................................... 19
5.27 Diffuser placement ...................................................................................................................... 19
5.28 Total work area components placement ..................................................................................... 19
5.29 Data plate(s) ............................................................................................................................... 19
5.30 Routine maintenance adjustment fixtures .................................................................................. 20
5.31 User-modified pass-throughs...................................................................................................... 20
6 Performance ......................................................................................................................................... 27
6.1 General ....................................................................................................................................... 27
6.2 Pressure decay / soap bubble / tracer gas leak ......................................................................... 27
6.3 HEPA/ULPA filter leak ................................................................................................................ 27
6.4 Noise level .................................................................................................................................. 27
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6.5 Lighting intensity ......................................................................................................................... 27
6.6 Vibration ...................................................................................................................................... 27
6.7 Personnel, product, and cross-contamination protection ........................................................... 28
6.8 Stability ....................................................................................................................................... 28
6.9 Downflow and inflow velocities ................................................................................................... 28
6.10 Inflow velocity.............................................................................................................................. 29
6.11 Airflow patterns test .................................................................................................................... 29
6.12 Drain spillage trough leakage ..................................................................................................... 29
6.13 Motor / blower performance ........................................................................................................ 29
6.14 Electrical safety ........................................................................................................................... 29
6.15 Performance data ....................................................................................................................... 30
6.16 Record maintenance ................................................................................................................... 30
6.17 Air velocity stability...................................................................................................................... 30
Normative Annex 1 Performance tests ..................................................................................................... 31
N-1.1 Pressure decay / soap bubble ................................................................................................. 31
N-1.2 HEPA/ULPA filter leak test ...................................................................................................... 32
N-1.3 Noise level test ........................................................................................................................ 34
N-1.4 Lighting intensity test ............................................................................................................... 35
N-1.5 Vibration test ............................................................................................................................ 35
N-1.6 Personnel, product, and cross-contamination protection (biological) tests ............................. 36
N-1.7 Downflow velocity .................................................................................................................... 47
N-1.8 Inflow velocity (face velocity) test ............................................................................................ 48
N-1.9 Airflow patterns test ................................................................................................................. 51
N-1.10 Drain spillage trough leakage test ........................................................................................... 53
N-1.11 Motor / blower performance..................................................................................................... 53
N-1.12 Cabinet airflow stability ............................................................................................................ 54
N-1.13 Canopy connection test ........................................................................................................... 56
Normative Annex 2...................................................................................................................................... 75
N-2.1 Method to verify fitness for use of potential direct inflow measurement devices .................... 75
Normative Annex 3 Nebulizer selection and calibration ........................................................................... 77
N-3.1 Selection .................................................................................................................................. 77
N-3.2 Calibration................................................................................................................................ 77
Normative Annex 4 Evaluation of chemical resistance and abrasion resistance of surfaces................... 81
N-4.1 Chemical resistance ................................................................................................................ 81
N-4.2 Abrasion resistance ................................................................................................................. 81
Normative Annex 5 Field tests .................................................................................................................. 83
N-5.1 Field certification preconditions and intervals.......................................................................... 83
N-5.2 Downflow velocity .................................................................................................................... 84
N-5.3 Inflow velocity (face velocity) test ............................................................................................ 86
N-5.4 Airflow patterns test ................................................................................................................. 90
N-5.5 HEPA/ULPA filter leak test ...................................................................................................... 91
N-5.6 Pressure decay / soap bubble ................................................................................................. 94
N-5.7 Site installation assessment tests ........................................................................................... 95
N-5.8 Electrical leakage and ground circuit resistance and polarity tests ......................................... 98
N-5.9 Lighting intensity test ............................................................................................................... 98
N-5.10 Vibration test ............................................................................................................................ 99
N-5.11 Noise level tests ...................................................................................................................... 99
N-5.12 Record of field certification .................................................................................................... 101
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Informative Annex 1 Biosafety cabinet selection, installation, use, lifespan, and decommissioning ...... 103
I-1.1 Institutional safety consultation.............................................................................................. 103
I-1.2 Risk assessment procedure .................................................................................................. 103
I-1.3 BSC Class and Type selection .............................................................................................. 108
I-1.4 Site review before BSC purchase ......................................................................................... 116
I-1.5 BSC arrival and certification .................................................................................................. 118
I-1.6 Cleaning and disinfection of BSC total work area ................................................................. 118
I-1.7 BSC use practices and procedures ....................................................................................... 120
I-1.8 Moving a permanently installed BSC .................................................................................... 124
I-1.9 BSC lifespan .......................................................................................................................... 125
I-1.10 Decommissioning process..................................................................................................... 125
I-1.11 Definitions .............................................................................................................................. 126
Informative Annex 2 .................................................................................................................................. 143
I-2.1 Recommended biosafety cabinet decontamination procedure ............................................. 143
I-2.2 Recommended HEPA/ULPA filter disposal procedures ........................................................ 159
Informative Annex 3 Recommended materials, finishes, and construction ............................................ 161
I-3.1 Sheet metal and finishes ....................................................................................................... 161
I-3.2 Glass...................................................................................................................................... 161
I-3.3 HEPA/ULPA filter gasket materials ....................................................................................... 161
I-3.4 HEPA/ULPA filter case – Type IC ......................................................................................... 161
I-3.5 Specifications......................................................................................................................... 162
I-3.6 Sealants ................................................................................................................................. 162
I-3.7 Fans ....................................................................................................................................... 162
I-3.8 Components and wiring ......................................................................................................... 162
Informative Annex 4 Reference standards and specifications pertinent to Class II biosafety cabinetry 163
I-4.1 Miscellaneous publications .................................................................................................... 163
I-4.2 Federal specifications ............................................................................................................ 164
I-4.3 Federal standards .................................................................................................................. 165
I-4.4 Military specifications ............................................................................................................. 165
Informative Annex 5 .................................................................................................................................. 167
I-5.1 Helium leak test ..................................................................................................................... 167
I-5.2 Sulfur hexafluoride (SF6) leak test ......................................................................................... 168
Informative Annex 6 Protocol for the validation of alternate biosafety cabinet decontaminating
methods and agents .............................................................................................................. 171
I-6.1 Introduction ............................................................................................................................ 171
I-6.2 Protocol.................................................................................................................................. 171
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Foreword 2
The purpose of this standard is to establish minimum requirements for materials, design, construction, and
performance of biosafety cabinetry that are designed to protect personnel, product, and the environment.
This standard details requirements for performance testing as well as field certification testing.
This edition includes the following revisions:
Issue 172
This revision updates language regarding the pressure decay test in Section 6.2 and Sections N-1.1.1.3
through N-1.1.1.5.
Issue 173
This revision adds definitions for cleanable, easily cleanable, and tubing restraint and affirms revised and
new language related to design and construction requirements throughout Section 5. It also removes former
Sections 5.7.1.3, 5.11, 5.12, 5.18.
Issue 174
This revision affirms revised language related to recommended preventative maintenance procedures in
Section N-5.1.
Issue 176
This revision affirms revised language related to the verification of the Secondary Inflow Method throughout
Section N-1.8.3 and N-5.3.3.
Issue 177
This revision affirms revised and new language related to noise level tests in Sections 6.4.2, N-1.3.3.c,
N-1.3.4, N-5.11.3.c, N-5.11.3.g, and N-5.11.4.
Issue 179
This revision affirms new language related to the requirements for canopy connections as new Section
5.29.3.
Issue 180
This revision corrects language regarding motor blower performance procedure Section N-1.11.3.c.
Issue 181
This revision updates and adds language regarding motor blower performance procedure Sections
N-1.11.2 and N-1.11.3.b.
2 The information contained in this foreword is not part of this American National Standard (ANS) and has not been
processed in accordance with ANSI’s requirements for an ANS. As such, this foreword may contain material that has
not been subjected to public review or a consensus process. In addition, it does not contain requirements necessary
for conformance to the standard.
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Issue 182
This revision updates language related to the requirements canopy connection acceptance criteria in
Section N-1.13.5.
Issue 186
This revision removes former Sections 6.17.1 and N-1.13.3.1 related to drop testing.
Issue 187
This revision updates power failure disconnection time from 1 h to 5 min in Section 6.17.2 and Section
N-1.12.3.2.
Issue 190
This revision updates language related to the personal protection test in Section N-1.6.3.1.
Issue 191
This revision affirms revised language related to control plate placement in Sections N-1.6.3.1.c and
N-1.6.4.1.e.
Issue 192
This revision updates a reference in Section 6.8.1 and removes previous Sections N-1.7 through N-1.7.2
relating to the resistance to overturning test.
Issue 194
This revision updates language related to the noise level acceptance criteria in Section N-5.11.4 and adds
Table N-5.1.
Issue 195
This revision updates language related to the installation and lifespan of BSCs throughout Sections I-1.3,
I-1.8, and I-1.9.
Issue 197
This revision updates and adds new language related to chemical resistance testing in Sections 4.5.2.1,
N-4.1.1, and N-4.1.2.
Issue 198
This revision adds new language related to replacement filters in Section 5.19.
The Interpretation Annex contains responses to interpretation requests. The responses will be published in
each version of the standard until such time that the interpretation response is no longer applicable.
This standard was developed by the NSF Joint Committee on Biosafety Cabinetry using the consensus
process described by the American National Standards Institute.
This standard and the accompanying text are intended for voluntary use by certifying organizations,
regulatory agencies, and/or manufacturers as a basis of providing assurances that adequate health
protection exists for covered products.
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Suggestions for improvement of this standard are welcome. This standard is maintained on a continuous
maintenance schedule and can be opened for comment at any time. Comments should be sent to:
Chair, Joint Committee on Biosafety Cabinetry at standards@nsf.org, or c/o NSF International, Standards
Department, P.O. Box 130140, Ann Arbor, Michigan 48113-0140, U.S.A.
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© 2024 NSF NSF/ANSI 49 – 2024
NSF/ANSI Standard
for Biosafety Cabinetry ―
Biosafety Cabinetry: Design, Construction,
Performance, and Field Certification
1 General
1.1 Scope
This standard applies to Class II (laminar flow) biosafety cabinetry designed to minimize hazards inherent
in work with agents assigned to Biosafety Levels 1, 2, 3, or 4. It also defines the tests that shall be passed
by such cabinetry to meet this standard. This standard includes basic requirements for the design,
construction, and performance of biosafety cabinets (BSCs) that are intended to provide personnel, product,
and environmental protection; reliable operation; durability and structural stability; cleanability; limitations
on noise level; illumination; vibration; and motor / blower performance.
1.2 Minimum requirements
Cabinets qualifying under this standard shall have passed all of the designated tests. Units with component
parts covered under existing NSF standards or criteria shall conform to those applicable requirements.
1.3 Variations in design and construction
Cabinetry varying in design, construction, or installation of accessory equipment may qualify under this
standard, if appropriate tests and investigations indicate that the equipment is durable and reliable, can be
cleaned and decontaminated, and performs in conformance to this standard. Such equipment shall meet
the requirements for materials and finishes in this standard.
Major modifications require appropriate tests for conformance. Major modifications include, but are not
limited to any of the following changes to the blower / motor(s): location, capacity, quantity, or automatic
airflow adjustment; size, or design, or both, of air plenums; position of high efficiency particulate air (HEPA)
/ ultra-low penetrating air (ULPA) filters; position or redesign of work surface; total work area intake and
exhaust air grilles; window placement or design; access opening size; location and size of exhaust port; the
visibility or audibility of the safety signaling; and built-in accessory equipment (centrifuges, ultraviolet [UV]
lighting, supports for intravenous drug container, arm rests, etc.). Major modifications also include changes
affecting the safe use of the cabinet including the ability to see, hear, and understand the required alarms.
Relocation of utility service equipment (electrical outlets, petcocks, etc.), the visual appearance of the
cabinet, or user interface(s), are not considered a major modification if other provisions of this standard are
not compromised.
2 Normative references
The following documents contain requirements that, by reference in this text, constitute requirements of this
standard. At the time of publication, the indicated editions were valid. All of the documents are subject to
revision and parties are encouraged to investigate the possibility of applying the most recent editions of the
documents indicated below.
