CRC handbook of laboratory Safety - Chapter 1 docx

Furr, A. Keith Ph.D. "Frontmatter"
CRC handbook of laboratory Safety
Edited by A. Keith Furr, Ph.D.
Boca Raton: CRC Press LLC,2000
© 2000 CRC Press LLC
FOREWORD
In rereading the foreword for the 4
th
edition, I noted the statement “There does not appear
to be much pending for the immediate future.” I was wrong. There have been significant changes
in many areas, such as respiratory protection, and dramatic changes in the Clean Air Act which
have made it difficult to use incineration as a means of disposing of hospital, medical, and
infectious waste. Perhaps the most significant change howev er has been in means of
communication, with the explosive growth of the Internet. This has placed a tremendous amount
of information available to anyone with a computer and a modem. Indeed, there is so much
information, one must be careful to select that which is useful and accurate. This resource has
greatly influenced preparation of this handbook.
At first glance, one might assume that little has changed in much of this edition. Again, this
would not be correct. The same general topics do remain for the most part, but several older and
now obsolete articles have been completely removed and replaced, either with new material on
the same subject or by completely new material, representing over a hundred pages. Where the
material may at first glance look familiar, please look more carefully. Every word on every page
has been scrutinized and there are literally hundreds of changes to bring the material up to date
or clarify the presentation.
There are new figures illustrating new material, and new tables. Full use of the Internet has
been made to make sure the information is as up-to-date as possible as of the end of the summer
of 1999. In addition to the usual journal articles as references, most topics now include Internet
references which were used and which I believe will be helpful.
I am pleased with this edition and believe it is the most authoritative of those for which I

have been responsible. One point I wish to close with and that is, safety in the laboratory is not
just a rigid adherence to regulatory standards and guidelines. It must take into account human
factors as well, and unfortunately the first part of the old adage “To err is human, to forgive is
divine” is all too true. Human nature being what it is, the vast proportion of breakdowns in
laboratory safety are due to human error, sometimes due to oversights but also sometimes due
to a feeling that it’s not important or individuals feel they will not make a mistake. This is
reflected throughout the handbook and reflects either my personal experience or observations.
I hope that no one is bothered by this intrusion.
I hope you will find the handbook useful as many have been kind enough to tell me they did
the previous editions.
© 2000 CRC Press LLC
THE EDITOR
A. Keith Furr, Ph.D., was, until his retirement in late 1994, Head of the Department of
Environmental Health and Safety at Virginia Polytechnic Institute and State University,
Blacksburg, Virginia, and Professor of Nuclear Science and Engineering. He received an A.B.
degree, cum laude, from Catawba College in 1954, an M.S. degree from Emory University in 1955,
and a Ph.D. from Duke University in 1962. From 1960 until 1971, he was in the Department of
Physics at VPI & SU where he attained the rank of Professor. In 1971, he transferred to
Engineering as Professor of Nuclear Science and Engineering. In 1975, he established the
Environmental Health and Safety Department at the University. A unique feature of this
department is that it eventually included a University volunteer rescue squad composed entirely
of students. In addition to other assignments, he was Director of the Nuclear Reactor Facility
and Head of the Neutron Activation Analysis Laboratory. During the early 1970s, he created an
undergraduate program in Radiation Safety and afterward he participated in a broad program in
Industrial Safety in the University's Department of Industrial and Systems Engineering. In recent
years, he played a leadership role in developing a unique program in correcting indoor air quality
problems in the University. He belongs to the Health Physics Society, the Campus Safety
Association, the National Safety Council, and the National Fire Protection Association. He has
published over 60 articles in professional journals, many in the area of environmental studies,
three encyclopedia articles and was editor and principal contributor to the two previous editions

of this handbook. After his retirement, he became a member of the advisory board of the
Laboratory Safety & Environmental Management Newsletter and Conference. He has
contributed numerous articles to the Newsletter. Dr. Furr has been active in working with public
bodies to develop programs that respond to environmental emergencies; that address the
disposal of hazardous materials, infectious wastes, and other solid wastes; and that are
environmentally and economically sound. He was Chair of the Montgomery County Local
Emergency Planning Committee and a member of the County Solid Waste and Recycling
Committee. He was also Chair of the Blacksburg Telecommunications Committee and of a group
of active Internet users called the Blacksburg Electronic Village Seniors.
He and his wife moved to Brooksville, Florida in May, 1998, where most of this handbook
was prepared.
© 2000 CRC Press LLC
THE CONTRIBUTORS
John W. Cure, III, Health Physics Consultation, Lynchburg, Virginia
Richard F. Desjardins, M.D., Wilmington, North Carolina
Lawrence G. Doucet, P.E., Doucet & Mainka, P.C., Peekskill, New York
Caldwell N. Dugan, Division of Institutional Resources, National Science Foundation,
Washington, D.C.
Frank A. Graf, Jr., Research Scientist, Westinghouse Hanford Company, Richland,
Washington
Scott A. Heider, Division of Institutional Resources, National Science Foundation,
Washington, D.C.
Harold Horowitz, Division of Institutional Resources, National Science Foundation,
Washington, D.C.
Alvin B. Kaufman, Litton Systems Divison, Litton Industries, Woodland Hills, California
Edwin N. Kaufman, Senior Scientist, Douglas Aircraft Co., Woodland Hills, California
David M. Moore, D.V.M., Director, Laboratory Animal Resources, College of Veterinary
Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia
Eric W. Spencer, Brown University, Providence, Rhode Island
William L. Sprout, M.D., Medical Consultant, Haskell Laboratory, E.I. du Pont de

Nemours & Co., Newark, Delaware
Norman V. Steere, Laboratory Safety and Design Consultant, Minneapolis, Minnesota
M.A. Trevino, M.D., Medical Director, Quimca Fluor, S.A. de C.V., Matamoros, Tamaulipas,
Mexico
Paul Woodruff, Environmental Resources Management, Inc., West Chester, Pennsylvania
Note that these are only those individuals specifically named in the text. As noted in the
Dedication, the actual number of contributors was much, much greater.
© 2000 CRC Press LLC
TABLE OF CONTENTS
FOREWORD
THE EDITOR
THE CONTRIBUTORS
CONTENTS
DEDICATION
Chapter 1
INTRODUCTION
I. LABORATORY SAFETY AS A COOPERATIVE RESPONSIBILITY
A. Human Resources
B. Legal Department
C. Purchasing Department
D. Facilities Department
E. Management
F. Organizational Structure
G. The Safety Department
1. Functions Relevant to Laboratories for the Safety Department
H. Departmental Responsibilities
1. Laboratory Responsibilities
Chapter 2
EMERGENCIES
A. Components of Emergency Preparedness

1. Initial Conditions
2. Facilities, Fixed and Movable Equipment
B. Institutional or Corporate Emergency Committee
C. Emergency Plan
1. Laboratory Emergency Plan
2. Organizational Emergency Plan
a. Emergency Plan Components
b. Emergency Equipment
c. Basic Emergency Procedures
D. Emergency Procedures for Selected Emergencies
1. Spills
2. Fire
3. Explosions
4. Toxic Air Quality
5. Radioactive and Contagious Biological Material Releases
a. Biological Accident
© 2000 CRC Press LLC
b. Radiation Incident
6. Multiple Class Emergencies
E. Artificial Respiration, Cardiopulmonary Resuscitation (CPR), and First Aid
1. Artificial Respiration
a. Artificial Respiration, Manual Method
b. Artificial Respiration, Mouth-to-Mouth Method
2. Cardiopulmonary Resuscitation
a. Initial Steps
b. Formal CPR Procedures
c. First Aid
Chapter 3
LABORATORY FACILITIES—DESIGN AND EQUIPMENT
I. LABORATORY DESIGN

A. Engineering and Architectural Principles
B. Building Codes and Regulatory Requirements
1. Building Classification
2. Types of Construction
C. Laboratory Classification
1. Program-Related Factors
2. Laboratory Class Characteristics
a. Low-Risk Facility
b. Moderate-Risk Facility
c. Substantial-Risk Facility
d. High-Risk Facility
D. Access
1. Exitways
a. Required Exits
b. Exit Capacity
c. Travel Distance
d. Corridors.
e. Stairs
f. Doors
g. Exit Signs, Lights, Emergency Power
h. Other Exitway Issues
E. Construction and Interior Finish
1. Construction Practices
2. Interior Finish
F. Ventilation
1. Quality of Supplied Air
G. Electrical Systems
1. Hazardous Locations
H. Plumbing
1. Sanitary System Materials

2. Back flow Prevention
I. Other Laboratory Utilities
J. Maintenance Factors
II. FIXED EQUIPMENT AND FURNITURE
A. Laboratory Furniture
1. Base Units and Work Tops
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2. Storage Cabinets
a. Flammable Material Storage
b. Cabinets for Drug Storage
c. Storage of Radioactive Materials
d. Corrosive Materials
e. Records Protection
B. Hoods
1. Factors Affecting Performance
a. Face Velocity
b. Construction Materials
c. Fume Hood Bases
d. Sashes
e. Internal Fixtures
2. Types of Chemical Fume Hoods
a. Conventional Fume Hood
b. Bypass Hood
c. Auxiliary Air Hoods
d. Walk-In Hood
e. Self-Contained Hoods
f. Hoods for Compliance with the Americans with Disabilities Act (ADA)
g. Other Modes of Exhaust
h. Perchloric Acid Hood
i. Radioisotope Fume Hood

j. Carcinogen Fume Hood
3. Exhaust Ducts
a. Materials
b. Dimensions
c. Fan Selection
4. Energy Management
5. Biological Safety Cabinets
a. Class I Cabinet
b. Class II Cabinets
c. Class III Cabinets (Glove Boxes)
d. HEPA Filters
e. Installation, Maintenance, and Certification
6. Built-In Safety Equipment
a. Eyewash Stations
b. Safety (Deluge) Showers
c. Fire Suppression Systems
d. Fire Detection and Alarm Systems
7. Other Fixed Equipment
III. CHEMICAL STORAGE ROOMS
A. Capacity
B. Construction Features
C. Ventilation
D. Fire Safety
IV. MOVABLE EQUIPMENT
A. Refrigeration Equipment
B. Ovens
C. Heating Baths
D. Stills
E. Kjeldahl Systems
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F. Autoclaves
G. Aerosol Generating Devices
V. ANIMAL LABORATORIES—SPECIAL REQUIREMENTS
A. Fixed Equipment in Animal Holding Facilities
1. Cage Washers
2. Autoclaves
3. Incinerator
B. Equipment for Animal Laboratories and Holding Areas
Chapter 4
LABORATORY OPERATIONS
I. GENERAL CONSIDERATIONS
II. OSHA LABORATORY SAFETY STANDARD
A. The Chemical Laboratory
B. Chemical Hygiene Plan
1. Goals
2. Organization
3. Training and Information Program
4. Medical Program
5. Laboratory Produced Chemicals
6. Record Keeping
7. Summary
III. OPERATIONAL PLANNING
A. Quantities
B. Sources
C. Material Safety Data Sheets
D. Purchase of Regulated Items
1. Radioisotopes
2. Controlled Substances (Drugs)
3. Etiologic Agents
4. Carcinogens

5. Explosives
6. Equipment
E. Free Materials
IV. PURCHASING OF ANIMALS
A. Introduction
1. Selection Criteria for Rodents and Rabbits
2. Laws Affecting Animal Purchasing
3. Transportation of Animals
4. Additional Laws Affecting Animal Purchase
V. STORAGE
A. Compatible Chemical Storage
B. Labeling
C. Regulated Materials
1. Explosives
2. Controlled Substances (Drugs)
3. Radioisotopes
4. Etiologic Agents
5. Carcinogens
D. Ethers
1. Formation of Peroxides
2. Detection and Estimation of Peroxides
© 2000 CRC Press LLC
3. Inhibition of Peroxides
4. Removal of Peroxides
E. Perchloric Acid
1. Perchloric Acid Storage
F. Flammable Liquids
G. Refrigeration Storage
H. Gas Cylinders
1. Bulk Storage

2. Laboratory Storage
3. Animal Food and Supply Storage
a. Animal Food
b. Supply Storage
c. Animal Carcass Storage
VI. HANDLING AND USE OF CHEMICALS: LABORATORY OPERATIONS
A. Physical Laboratory Conditions
1. Organization of the Laboratory
2. Eating, Studying, and Other Social Activities
3. Maintenance
4. Housekeeping
5. Signs and Symbols
B. Working Procedures
1. Protection Against Explosions
a. Personal Protective Equipment
b. Summary
2. Corrosive Chemicals
3. High-Energy Oxidizers
4. Perchloric Acid
a. Recommendations for the Safe Handling of Perchloric Acid
b. Decontamination/Removal of a Perchloric Acid Fume Hood System
c. Procedures for Decommissioning a Perchloric Acid Hood
5. Ethers
6. Flammable Solvents
a. Flammable Hazards
7. Reactive Metals
8. Mercury
a. Absorption of Mercury by the Body
b. Excretion of Mercury
c. Control Measures

d. Exposure Reduction
e. Monitoring
f. Spill Control Measures
g. Ventilation
h. Medical Surveillance
9. Hydrofluoric Acid
a. Treatment to Exposure
10. Hydrogen Cyanide
11. Fluorine Gas
12. General Safety for Hazardous Gas Research
13. Some Hazardous Gases
14. Cryogenic Safety
a. Hazards
b. General Precautions
© 2000 CRC Press LLC
c. Storage
d. Addendum to Section 14
15. Cold Traps
a. Virtual Leaks
b. Safety Precautions
16. Care and Use of Electrical Systems
a. Electrical Shock
b. Resistive Heating
c. Spark Ignition Sources
17. Glassware
a. Other Safety Problems
b. Glassware Cleaning
18. Unattended Operating Equipment
a. Laboratory Distillations
b. Other Laboratory Operations

VII. HAZARD AWARENESS (RIGHT-TO-KNOW)
A. Basic Requirements
B. Written Hazard Communication Program
1. Personnel Lists
2. Chemical List
3. Labeling
4. Material Safety Data Sheets
5. Employee Training and Information
VIII. HEALTH EFFECTS
A. Exposure Limits
B. Environmental Monitoring
C. Modes of Exposure
1. Means of Contact and Entry of Toxic Agents
a. Skin Contact
b. Inhalation
c. Biologic Aspects of Particulate Matter
d. Biologic Aspects of Gases and Vapors
e. Ingestion
D. Health Assurance Program
1. Basis of a Health Assurance Program
a. Health Assurance Medical Departments
2. A Health Assurance Program
a. Participation
b. Medical and Work Histories
c. Pre-Placement Examination
d. Reexamination
e. Utilization of Results
f. Physician Training
g. Records
h. CPR and First Aid Training

i. Vaccinations
E. The OSHA Bloodborne Pathogen Standard: Infection from Work with
Human Specimens
1. Basic Provisions
2. Exposure Control Plan
a. Exposure Determination
b. Implementation of the Exposure Control Plan
© 2000 CRC Press LLC
3. HIV Infection: Detection, Initial Management, and Referral
4. Hepatitis A and B
a. Hepatitis A
b. Hepatitis B
5. Zoonotic Diseases
a. Modes of Transmission
b. Routes of Exposure
c. Allergies
d. Waste Collection and Storage
e. Bedding
F. Reproductive Hazards
G. Regulated and Potential Carcinogens
1. Carcinogens (Ethylene Oxide)
H. Neurological Hazards of Solvents
IX. SPILLS AND EMERGENCIES
A. Small- to Moderate-Scale Spills
1. Spill Response Procedures
B. Large-Scale Releases of Chemicals
1. Hazardous Waste Operations and Emergency Response
2. Safety and Health Program
a. Organizational Structure
b. Comprehensive Work Plan

c. Site-Specific Safety and Health Plan
d. Hazard Identification and Relevant Information
e. Personal Protective Equipment
f. Monitoring
3. Hazard Communication Program
a. Training
4. Medical Surveillance Program
5. Engineering Controls, Work Practices, and Personal Protective
Equipment for Employee Protection
6. SARA Title III, Community-Right-to-Know
X. CHEMICAL WASTES
A. Resource Conservation and Recovery Act
1. Definition of a Hazardous Waste
2. Requirements for Generators of Hazardous Waste
a. Basics of Compliance with HM-181
b. Container Labels
c. Local Waste Management
3. Record Keeping Required of the Generator
4. Personnel Training
5. Preparedness and Prevention
6. Contingency Plan
7. Emergency Procedures
B. Practical Hazardous Waste Management Program
1. Internal Waste Management Organization
2. Reduction of Hazardous Waste Volume
3. Waste Collection
4. Packaging
5. Characterization of the Waste
6. Packing of Waste for Shipment
7. Restrictions on Wastes

© 2000 CRC Press LLC
8. Shipping Waste
9. Landfill Disposal
10.Incineration of Hazardous Waste
C. State-of-the-Art Hospital and Institutional Waste Incineration, Selection,
Procurement, and Operations
1. Introduction
2. Incineration Technologies
3. Sizing and Rating
4. Primary Combustion Chambers
a. Heat Release Rates
b. Burning Rates
c. Secondary Combustion Chambers
d. Shapes and Configurations
5. Selection and Design Factors
a. Capacity Determination
b. Burning Rate vs. Charging Rate
6. Incinerator System Auxiliaries
a. Residue Removal and Handling Systems
b. Waste Heat Recovery
c. Chemical Waste Incineration
d. Emission Control System
e. Success Data
7. Incineration Performance and Procurement
a. Fundamental Reasons for Poor Performance
8. Key Step
9. Recommended Procurement Steps
D. Hospital, Medical, and Infectious Waste
A. Incineration in Compliance With the Clean Air Act
A. Incinerator Classes

B. Other Regulatory Requirements for New and Existing Units
B. Alternative Technologies
C. Storage and Transportation of Medical and Infectious Waste
XI. LABORATORY CLOSEOUT PROCEDURES
Chapter 5
NONCHEMICAL LABORATORIES
I. INTRODUCTION
II. RADIOISOTOPE LABORATORIES
A. Brief Summary of Atomic and Nuclear Concepts
B. Radiation Concerns
C. Natural Radioactivity
D. Basic Concepts
1. The Atom and Types of Decay
2. The Fission Process
3. Radioactive Decay
4. Units of Activity
5. Interaction of Radiation with Matter
a. Alphas
b. Betas
c. Gammas
6. Units of Exposure and Dose
© 2000 CRC Press LLC
E. Licensing
1. Radiation Safety Committees
F. Radiation Protection, Discussion, and Definitions
1. Selected Definitions
2. Selected Radiation Protection Standards from 10 CFR Part 20
a. Occupational Limits for Adult Employees
b. Occupational Limits for Minors (Under 18) and to an Embryo/Fetus
c. Dose Limits for Individual Members of the Public

d. Surveys and Monitoring
e. Notification of Incidents
3. EPA, National Emission Standards for Hazardous Air Pollutants,
Radionuclides
G. Radioisotope Facilities and Practices
1. Radiation Working Areas
2. External Radiation Exposure Areas
3. Areas with Possible Internal Exposures
H. Material Control Procedures
1. Ordering and Receipt of Materials
I. Operations
1. Reduction of Exposures, ALARA Program
a. Selection of Radioisotopes
b. Shielding
c. Distance
d. Time
e. Quantity
f. Example of Time and Distance
g. Internal Dose Limiting
h. Methods of Monitoring Personnel Exposures
i. Bioassays
j. Radiation Surveys
k. Measurement of Airborne Activities
l. Fixed and Loose Surface Contamination
m. Radioactive Waste Disposal
n. Individual Rights and Responsibilities
J. Radon
K. Acute Radiation Syndrome
III. X-RAY FACILITIES
A. Generation of X-Rays

B. Types of Machines
1. Diagnostic Machines
a Primary Beam Shielding
b. Scattered Radiation
c. Leakage Radiation
d. Exposure to Users
2. X-Ray Quality Control in Medical X-Ray Laboratories
3. Open-Beam Analytical Machines
4. Closed-Beam Analytical Systems
5. Cabinet X-Ray Systems
6. Miscellaneous Systems
IV. NONIONIZING RADIATION
V. LASER LABORATORIES
A. Protective Procedures for Class IIIb and Class IV Lasers
© 2000 CRC Press LLC
B. Eye Protection
C. Medical Surveillance
VI. MICROBIOLOGICAL AND BIOMEDICAL LABORATORIES
A. Introduction
B. Laboratory Safety and Hazard Communication Standards
C. Laboratory Director
D. Miscellaneous Safety Practices
1. Laboratory Line of Authority
2. Spills and Emergencies Involving Chemically Dangerous
Materials
3. Emergency Equipment for Non-biohazardous Spills
4. Evacuation
E. Accidents and Spills of Biohazardous Materials
F. Generation of Aerosols
G. Infectious Waste

H. Laboratory Facilities — Design and Equipment
I. Biosafety Levels
1. Biosafety Level 1
a. Standard Microbiological Practices
b. Special Practices
c. Safety Equipment (Primary Barriers)
d. Laboratory Facilities (Secondary Barriers)
2. Biosafety Level 2
a. Standard Microbiological Practices
b. Special Practices
c. Safety Equipment (Primary Barriers)
d. Laboratory Facilities (Secondary Barriers)
3. Biosafety Level 3
a. Standard Microbiological Practices
b. Special Practices
c. Safety Equipment (Primary Barriers)
d. Laboratory Facilities (Secondary Barriers)
4. Biological Safety Level
a. Standard Microbiological
b. Special Practices
c. Safety Equipment (Primary Barriers)
d. Laboratory Facility (Secondary Barriers)
J. Vertebrate Animal Biosafety Level Criteria
1. Animal Biosafety Level 1
a. Standard Practices
b. Special Practices ABSL 1
c. Safety Equipment (Primary Barriers) ABSL 1
d. Animal Facilities (Secondary Barriers) ABSL 1
2. Animal Biosafety Level 2
a. Standard Practices

b. Special Practices ABSL 2
c. Safety Equipment (Primary Barriers) ABSL 2
d. Animal Facilities (Secondary Barriers) ABSL 2
3. Animal Biosafety Level 3
a. Standard Practices
b. Special Practices ABSL 3
c. Safety Equipment (Primary Barriers) ABSL 3
© 2000 CRC Press LLC
d. Animal Facilities (Secondary Barriers) ABSL 3
4. Animal Biosafety Level 4
a. Standard Practices
b. Special Practices ABSL 4
c. Safety Equipment (Primary Barriers
d. Animal Facility (Secondary Barriers) ABSL 4
K. Recommended Biosafety Levels
1. Agent Summaries
a. Parasitic Agents
b. Fungal Agents
c. Bacterial Agents
d. Rickettisial Agents
e. Viral Agents
f. Arboviruses Assigned to Biosafety Level 2.
g. Vaccine Strains of Biosafety Level 3/4 Viruses which
May be Handled at Biosafety Level 22
h. Arboviruses and Certain Other Viruses Assigned to Biosafety
Level 3 (on the basis of insufficient experience)
i. Arboviruses and Certain Other Viruses Assigned to Biosafety
Level 3
j. Arboviruses, Arenaviruses, and Filoviruses Assigned to Biosafety
Level 4

k. Restricted Animal Pathogens.
VII. RECOMBINANT DNA LABORATORIES
A. Section I-B. Definition of Recombinant DNA Molecules
B. I-C. General Applicability
C. I-D. Compliance with the NIH Guidelines
D. Section II, Safety Considerations
1. IIA-1. Risk Groups
2. IIA-2. Risk Assessment
3. IIA-3. Risk Assessment Factors
E. II-B. Containment
F. Section III. Experiments Covered by the NIH Guidelines
1. Section III-A.
*
Experiments that Require Institutional Biosafety
Committee Approval, RAC Review, and NIH Director Approval
Before Initiation
a. III-A-1, Major Actions Under the Guidelines
b. Section III-A-1-a.
2. Section III-B. Experiments That Require NIH/ORDA and Institutional
Biosafety Committee Approval Before Initiation
a. Section III-B-1. Experiments Involving the Cloning of Toxin
Molecules with LD
50
Less than 100 Nanograms per Kilogram
Body Weight
3. Section Ill-C. Experiments that Require Institutional Biosafety
Committee and Institutional Review Board Approvals and
NIH/ORDA Registration Before Initiation
a. Section Ill-C-1. Experiments Involving the Deliberate Transfer
of Recombinant DNA or DNA or RNA Derived from Recombinant

DNA into One or More Human Subjects
4. Section III-D. Experiments that Require Institutional Biosafety
Committee Approval Before Initiation
© 2000 CRC Press LLC
a. Section III-D-1. Experiments Using Risk Group 2, Risk Group 3,
Risk Group 4, or Restricted Agents as Host-Vector Systems
b. Section III-D-2. Experiments in Which DNA From Risk Group 2,
Risk Group 3, Risk Group 4, or Restricted Agents is Cloned
into Nonpathogenic Prokaryotic or Lower Eukaryotic Host-Vector
Systems
c. Section III-D-3. Experiments Involving the Use of Infectious DNA
or RNA Viruses or Defective DNA or RNA Viruses in the Presence
of Helper Virus in Tissue Culture Systems
d. Section III-D-4. Experiments Involving Whole Animals
e. Section III-D-5. Experiments Involving Whole Plants
f. Section III-D-6. Experiments Involving More than 10 Liters of
Culture
g. Section III-E. Experiments that Require Institutional Biosafety
Committee Notice Simultaneous with Initiation
h. Section III-F. Exempt Experiments
G. Section IV. Roles and Responsibilities
1. Section IV-A. Policy
2. Section IV-B. Responsibilities of the Institution
a. Section IV-B-1. General Information
3. Section IV-B-2. Institutional Biosafety Committee (IBC
a. Section IV-B-2-a. Membership and Procedures
b. Section IV-B-2-b. Functions
4. Section IV-B-3. Biological Safety Officer (BSO)
a. Section IV-B-3-a.
b. Section IV-B-3-b

c. Section IV-B-3-c
4. Section IV-B-7. Principal Investigator (P1)
a. Section IV-B-7-a. General Responsibilities
b. Section IV-B-7-b. Submissions by the Principal Investigator to
NIH/ORDA
c. Section IV-B-7-c. Submissions by the Principal Investigator
to the Institutional Biosafety Committee
d. Section IV-B-7-d. Responsibilities of the Principal Investigator
Prior to Initiating Research
e. Section IV-B-7-e. Responsibilities of the Principal Investigator
During the Conduct of the Research
H. Section IV-D. Voluntary Compliance
1. Section IV-D-1. Basic Policy - Voluntary Compliance
2. Section IV-D-2. Institutional Biosafety Committee Approval-Voluntary
Compliance
3. Section IV-D-3. Certification of Host-Vector Systems-Voluntary
Compliance
4. Section IV-D-4. Requests for Exemptions and Approvals - Voluntary
Compliance
5. Section IV-D-5. Protection of Proprietary Data - Voluntary Compliance
a. Section IV-D-5-a. General
b. Section IV-D-5-b. Pre-submission Review
F. Appendices
1. Appendix A. Exemptions Under Section III-f-5-Sublists of Natural
© 2000 CRC Press LLC
Exchangers
a. Appendix A-I. Sublist A
b. Appendix A-II. Sublist B
c. Appendix A-III. Sublist C
d. Appendix A-IV. Sublist D

e. Appendix A-V. Sublist E
f. Appendix A-VI. Sublist F
2. Appendix B. Classification of Human Etiologic Agents on the Basis
of Hazard
a. Appendix B - Table 1. Basis for the Classification of Biohazardous
Agents by Risk Group (RG)
b. Appendix B-I. Risk Group 1 (RG1) Agents
c. Appendix B-II. Risk Group 2 (RG2) Agents
d. Appendix B-II-A. Risk Group 2 (RG2) - Bacterial Agents Including
Chlamydia
e. Appendix B-II-B. Risk Group 2 (RG2) - Fungal Agents
f. Appendix B-II-C. Risk Group 2 (RG2) - Parasitic Agents
g. Appendix B-II-D. Risk Group 2 (RG2) - Viruses
h. Appendix B-III. Risk Group 3 (RG3) Agents
i. Appendix B-III-A Risk Group 3 (RG3)- Bacterial Agents Including
Rickettsia
j. Appendix B-III-B. Risk Group 3 (RG3) - Fungal Agents
k. Appendix B-III-C. Risk Group 4 (RG3) - Parasitic Agents
l. Appendix B-III-D. Risk Group 3 (RG3) - Viruses and Prions
m. Appendix B-IV. Risk Group 4 (RG4) Agents
n. Appendix B-IV-A. Risk Group 4 (RG4) - Bacterial Agents
o. Appendix B-IV-B. Risk Group 4 (RG4) - Fungal Agents
p. Appendix B-IV-C. Risk Group 4 (RG4) - Parasitic Agents
q. Appendix B-IV-D. Risk Group 4 (RG4) - Viral Agents
r. Appendix B-V. Animal Viral Etiologic Agents in Common Use
s. Appendix B-V-1. Murine Retroviral Vectors
3. Appendix C. Exemptions Under Section III-F-6
a. Appendix C-I. Recombinant DNA in Tissue Culture
b. Appendix C-I-A. Exceptions
c. Appendix C-II. Escherichia coli K-12 Host-Vector Systems

d. Appendix C-II-A. Exceptions
e. Appendix C-III. Saccharomyces Host-Vector Systems
f. Appendix C-III-A. Exceptions
g. Appendix C-IV. Bacillus subtilis or Bacillus licheniformis
Host-Vector Systems
h. Appendix C-IV-A. Exceptions
i. Appendix C-V. Extrachromosomal Elements of Gram Positive
Organisms
j. Appendix C-VI. The Purchase or Transfer of Transgenic Rodents
k. Appendix C-VII-A and A-1.
l. Appendix C-VII-B.
m. Appendix C-VII-C.
n. Appendix C-VII-D.
o. Appendix C-VII-E.
4. Appendix D. Major Actions Taken under the NIH Guidelines
5. Appendix E. Certified Host-vector Systems
© 2000 CRC Press LLC
a. Appendix E-I. Bacillus subtilis
b. Appendix E-I-A. Bacillus subtilis Host-Vector 1 Systems
c. Appendix E-I-B. Bacillus subtilis Host-Vector 2 Systems
d. Appendix E-II. Saccharomyces cerevisiae
e. Appendix E-II-A. Saccharomyces cerevisiae Host-Vector 2 Systems
f. Appendix E-III. Escherichia coli
g. Appendix E-III-A. Escherichia coli (EK2) Plasmid Systems
h. Appendix E-III-B. Escherichia coli (EK2) Bacteriophage Systems
i. Appendix E-IV. Neurospora crassa
j. Appendix E-IV-A. Neurospora crassa Host-Vector 1 Systems
k. Appendix E-V. Streptomyces
l. Appendix E-V-A. Streptomyces Host-Vector 1 Systems
m. Appendix E-VI. Pseudomonas putida

n. Appendix E-VI-A. Pseudomonas putida Host-Vector 1 Systems
6. Appendix F. Containment Conditions for Cloning of Genes Coding
for the Biosynthesis of Molecules Toxic for Vertebrates
a. Appendix F-I. General Information
b. Appendix F-II. Cloning of Toxin Molecule Genes in
Escherichia coli K-12
c. Appendix F-II-A.
d. Appendix F-II-B.
e. Appendix F-II-C.
f. Appendix F-III. Cloning of Toxic Molecule Genes in Organisms
Other Than Escherichia coli K
g. Appendix F-IV. Specific Approvals
7. Physical Containment
8. Appendix H. Shipment
a. Appendix H-I.
b. Appendix H-II.
c. Appendix H-III. Additional Resources
9. Appendix I. Biological Containment
a. Appendix I-I. Levels of Biological Containment
b. Appendix I-I-A. Host-Vector 1 Systems
c. Appendix I-I-B. Host-Vector 2 Systems
d. Appendix I-II. Certification of Host-Vector Systems
e. Appendix I-II-B. Data to be Submitted for Certification
10. Appendix K. Physical Containment for Large Scale Uses
of Organisms Containing Recombinant DNA Molecules
a. Appendix K-I. Selection of Physical Containment Levels
b. Appendix K-II. Good Large Scale Practice (GLSP)
11. Appendix M. Points to Consider in the Design and Submission of
Protocols for the Transfer of Recombinant DNA Molecules into
One or More Human Subjects (Points to Consider)

12. Appendix M-III. Informed Consent
a. Appendix M-III-A. Communication About the Study to Potential
Participants
b. Appendix M-III-B-1. General Requirements of Human Subjects
Research
c. Appendix M-III-B-2. Specific Requirements of Gene Transfer
Research
d. Appendix M-IV. Privacy and Confidentiality
© 2000 CRC Press LLC
13. Appendix P. Physical and Biological Containment for Recombinant
DNA Research Involving Plants
a. Appendix P-I. General Plant Biosafety Levels
b. Biological Containment Practices
14. Appendix Q. Physical and Biological Containment for Recombinant
DNA Research Involving Animals
a. Appendix Q-l. General Considerations
b. Appendix Q-I-A. Containment Levels
c. Appendix Q-l-B. Disposal of Animals (BL1 -N through BL4-N)
VIII. Research Animal Care and Handling
A. Introduction
B. Laws and Regulations Relating to Animal Care and Use
1. Animal Welfare Act
2. The Good Laboratory Practices Act
3. The Guide for the Care and Use of Laboratory Animals
4. Public Health Service Policies
5. Voluntary Regulations
C. Personnel
D. Animal Holding Facilities
E. Animal Care and Handling
F. Human Health Monitoring

Chapter 6
PERSONAL PROTECTIVE EQUIPMENT
I. INTRODUCTION
II. RESPIRATORY PROTECTION
A. “Dust” Masks
B. Half-Face Cartridge Respirators
C. Full-Face Respirators
D. Air-Supplied Respirators
E. OSHA 29 CFR 1910.134 (Slightly Abridged)
III. EYE PROTECTION
A. Chemical Splash Goggles
B. Safety Spectacles
C. Contact Lenses
IV. MATERIALS FOR PROTECTIVE APPAREL
A Recommended Information Sources
B. Overview of Chemical Protective Clothing
V. HEARING PROTECTION
VI. FIRST AID KITS
VII.OTHER PERSONAL PROTECTIVE EQUIPMENT
Appendix: Laboratory Checklist
© 2000 CRC Press LLC
DEDICATION
There are many individuals who must be thanked for their assistance with this book. As
with the 4
th
edition, my son Rob Furr who assisted me with solving technical problems.
Individuals such as John Cure, Lawrence Doucet, and David Moore who contributed material
for the 4
th
edition that has been carried over to this edition. Many other individuals who gave

lectures at meetings and with whom I talked at these meetings, whose insights and actual
knowledge found their way into the material. I hope that I’ve acknowledged all of these at the
relevant points in the text, by means of either a mention in the text or as a reference. I may
have missed some inadvertently, and if so, I hope that they accept my sincere apologies.
One group which I cannot overlook, but whom I cannot specifically name in most cases are
those individuals who compiled the numerous standards and guidelines to which I have
lavishly referred in this book. There are those who complain about regulatory standards but
one must have a mark at which to aim. There are also numerous persons and organizations
who have made information available on the Internet and this information has been extremely
helpful.
I must single out my wife for this edition for even more thanks than in the previous ones.
Not only did she tolerate the chaos and confusion alluded to in the last edition, but we
moved during the course of preparing this edition from a home in which we had lived for 30
years. She assumed much of the burden of that move and getting the new home organized.
In addition, she read every word of this edition to proofread it and make suggestions to
improve its clarity. The publisher also proofread the manuscript and eliminated still more
errors. I now more fully appreciate proofreaders! However, I assume full responsibility for
any residual errors since I followed up the previous reviews with a final review. I hope there
are none but I am sure some may still lurk within the pages.
Finally, I want to thank the purchasers of previous editions for their support. If they had
not made the 3
rd
and 4
th
editions a success, this one would not have been done.
Furr, A. Keith Ph.D. "INTRODUCTION"
CRC Handbook of Laboratory Safety
Edited by A. Keith Furr, Ph.D.
Boca Raton: CRC Press LLC,2000
©2000 CRC Press LLC

Chapter 1
INTRODUCTION
Safety and health programs for industrial organizations began to be significant early in the
20th century, and since then, there were slow but steady improvements in the industrial working
environment, until, in the early 1970s, the Federal Occupational Health and Safety Act was
passed as a national program to establish minimum standards for safety for industrial workers.
In order to comply with this Act, corporations in the United States have had to create formal
safety programs, and usually, internal safety organizations to comply with the regulations. The
more enlightened of these firms, recognizing the value to themselves and their employees of
keeping employees safe and well, strongly support these internal units. Smaller firms sometimes
have made a lesser commitment proportionally to safety, although there are increasingly fewer
exceptions. The standards incorporated in the regulations originally were based on previously
developed industrial standards created by various organizations. Some important groups were
not covered for a time, including public employees and effectively laboratory research workers.
For the latter group, the standards, based on industrial processes, simply did not translate well
to a laboratory scale environment. Many industrial organizations proceeded to develop their
own safety programs for laboratory personnel that were quite effective. Progressive universities
and colleges have created good safety programs, but again the strengths of these programs until
relatively recently were in the crafts and maintenance staff. Because of unique features of the
academic environment, safety programs affecting research operations typically were less
comprehensive than in industry. Until recently, when the OSHA standard for laboratory facilities
was enacted, even some larger educational institutions, and a very high percentage of moderate
to smaller schools have had minimal safety programs for their scientific employees. In conse-
quence, safety may not have been stressed as much as it should have been in academic research
institutions. In some schools, students can still graduate without having received any formal
safety training, or in some cases without even being exposed to good safety practices. Since
attitudes, once established, are very difficult to change, the attitude that safety is of secondary
importance still may be carried over into professional careers for a significant fraction of current
laboratory research personnel. Fortunately, this scenario is changing for the better as individuals
become more aware of the consequences of not doing things well.

Over a period of years, the regulatory environment has grown to encompass more of the
research environment, beginning with the users of radioactive materials in the 1940s and 1950s.
If an organization used radioactive materials, it was required to have a radiation safety officer or
committee, because these were mandated by the Nuclear Regulatory Commission (NRC), or its
©2000 CRC Press LLC
antecedents, under their licensing regulations. Approximately half the states have assumed this
responsibility as a surrogate for the NRC. In recent years, additional regulations or guidelines
have been passed explicitly covering laboratory operations affecting the disposal of chemicals,
infectious waste, exposure to human blood, tissue and other body fluids, the use of animals and
human subjects in research, and work involving recombinant DNA. Concern about risks to
workers and the environment from chemicals led to regulations requiring hazard information to
be given to individuals exposed to chemicals, and to local communities, where the potential
exposure to hazardous chemical incidents existed. These last regulations covered industrial
operations as well as research personnel. Finally, in 1991, OSHA passed a standard specifically
designed for the laboratory environment which required operators of research laboratories to
provide laboratory employees protection equivalent to that enjoyed by industrial workers
covered by the OSHA General Industry Standard. This laboratory oriented standard is a
performance standard, which is a virtual necessity in view of the tremendous variation in
laboratory operations. Although it provides flexibility, it is no less demanding in the net safety
provided laboratory personnel than specific standards. Additional hazard specific standards
have also been established. The 1992 Bloodborne Pathogen standard resulting from concerns
about exposures to AIDS and hepatitis probably affects more laboratory workers than
emergency care providers, the group that most persons immediately think of in the context of
accidental exposure to blood, tissue, and other bodily fluids. Infectious waste rules in force in
many states affect laboratories performing basic research in the life sciences as well as medical
facilities. The Americans With Disabilities Act has many implications for the laboratory
environment, although it too was intended to be applicable to employees in all occupations. Part
of the difficulty in developing an effective laboratory safety program lies in the nature of
laboratories. Research activities conducted within laboratories normally are extremely varied and
change frequently. The processes and materials in use may present unidentified problems.

Research materials may be being synthesized for the first time or may be being used in novel
ways. Flammable solvents are probably the most common class of chemicals in use, and there
are ample sources of ignition in most laboratories. Because of the changing needs in the
laboratory and the scale of most reactions over short periods, even a well-managed facility tends
to accumulate a large and varied inventory of partially used containers of chemicals. In one
moderate, not atypical, research laboratory at the author’s institution a recent inventory found
1041 different chemicals on the shelves, only a handful of which were curren tly in use or
anticipated to be in use in the near future. Many of these will never be used since research
personnel no longer trust their quality. Laboratory equipment is often fabricated or modified
within the laboratory or in an instrument shop maintained by the facility. Devices manufactured
or modified locally obviously are extremely unlikely to have been tested by any certified safety
testing organization. Laboratory facilities that may have been well designed for their intended
initial use may easily become wholly inadequate in terms of electrical services, ventilation, or
special equipment such as hoods, as programs change or as new occupants move into the
space. Subsequent changes are more often made solely to accommodate more activities rather
than to improve safety. It is also quite possible that a laboratory was not designed properly
initially. Relatively few architectural firms appear to really know how to design laboratories,
although this situation is definitely improving. Engineering factors are often the ones sacrificed
to achieve other goals. Buildings and spaces originally built to house classrooms in a university
are regularly converted to serve expanding research programs. These converted spaces rarely
serve well as laboratory structures without substantial and difficult to obtain sums being needed
to renovate them. Even with the best of intentions, physical solutions to safety problems that
may work well in large-scale industrial processes are often extremely difficult to scale to the
laboratory environment and alternatives may be expensive.
Individuals managing laboratories are usually very capable persons who tend to be strongly
goal oriented, often to the unintended exclusion of other factors. Scientists designing an
experiment may inadvertently neglect some peripheral factors affecting safety because they are
not directly applicable to attaining the research goal. They also may not remember that the
technicians or students under them may not have the years of experience and training that
research directors do. They may not evaluate all of the consequences if some item of equipment